U.S. patent application number 09/984490 was filed with the patent office on 2003-04-03 for 123 human secreted proteins.
Invention is credited to Brewer, Laurie A., Ebner, Reinhard, Fischer, Carrie L., Kyaw, Hla, LaFleur, David W., Li, Yi, Moore, Paul A., Olsen, Henrik S., Rosen, Craig A., Ruben, Steven M., Shi, Yanggu, Soppet, Daniel R., Zeng, Zhizhen.
Application Number | 20030064412 09/984490 |
Document ID | / |
Family ID | 27586903 |
Filed Date | 2003-04-03 |
United States Patent
Application |
20030064412 |
Kind Code |
A1 |
Fischer, Carrie L. ; et
al. |
April 3, 2003 |
123 human secreted proteins
Abstract
The present invention relates to novel human secreted proteins
and isolated nucleic acids containing the coding regions of the
genes encoding such proteins. Also provided are vectors, host
cells, antibodies, and recombinant methods for producing human
secreted proteins. The invention further relates to diagnostic and
therapeutic methods useful for diagnosing and treating disorders
related to these novel human secreted proteins.
Inventors: |
Fischer, Carrie L.; (Burke,
VA) ; Rosen, Craig A.; (Laytonsville, MD) ;
Soppet, Daniel R.; (Centreville, VA) ; Ruben, Steven
M.; (Olney, MD) ; Kyaw, Hla; (Frederick,
MD) ; Li, Yi; (Sunnyvale, CA) ; Zeng,
Zhizhen; (Lansdale, PA) ; LaFleur, David W.;
(Washington, DC) ; Moore, Paul A.; (Germantown,
MD) ; Shi, Yanggu; (Gaithersburg, MD) ; Olsen,
Henrik S.; (Gaithersburg, MD) ; Ebner, Reinhard;
(Gaithersburg, MD) ; Brewer, Laurie A.; (St. Paul,
MN) |
Correspondence
Address: |
HUMAN GENOME SCIENCES INC
9410 KEY WEST AVENUE
ROCKVILLE
MD
20850
|
Family ID: |
27586903 |
Appl. No.: |
09/984490 |
Filed: |
October 30, 2001 |
Related U.S. Patent Documents
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09984490 |
Oct 30, 2001 |
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09227357 |
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09227357 |
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60051926 |
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60052793 |
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Current U.S.
Class: |
435/7.9 ;
530/388.15; 530/391.1 |
Current CPC
Class: |
G01N 33/5011 20130101;
C12N 9/00 20130101; G01N 33/5047 20130101; A61K 38/00 20130101;
G01N 33/5058 20130101; C07K 14/47 20130101; G01N 33/6842 20130101;
A61P 7/00 20180101; A61P 37/00 20180101; A61P 29/00 20180101; A61P
25/00 20180101; G01N 33/5041 20130101; G01N 33/505 20130101; G01N
33/5008 20130101; G01N 33/68 20130101; G01N 2500/00 20130101 |
Class at
Publication: |
435/7.9 ;
530/391.1; 530/388.15 |
International
Class: |
G01N 033/53; G01N
033/542; C07K 016/46 |
Claims
What is claimed is:
1. An isolated antibody or portion thereof that specifically binds
to a protein whose sequence consists of amino acid residues 21 to
116 of SEQ ID NO:238.
2. The isolated antibody or portion thereof of claim 1, wherein
said protein specifically bound by said antibody or portion thereof
is glycosylated.
3. The isolated antibody or portion thereof of claim 1 which is a
monoclonal antibody.
4. The isolated antibody or portion thereof of claim 1 which is a
polyclonal antibody.
5. The isolated antibody or portion thereof of claim 1 which is a
chimeric antibody.
6. The isolated antibody or portion thereof of claim 1 which is a
humanized antibody.
7. The isolated antibody or portion thereof of claim 1 which is a
human antibody.
8. The isolated antibody or portion thereof of claim 1 which is a
Fab fragment.
9. The isolated antibody or portion thereof of claim 1 which is
labeled.
10. The isolated antibody of claim 9, wherein the label is selected
from the group consisting of: (a) an enzyme label; (b) a
radioisotope; and (c) a fluorescent label.
11. An isolated cell that produces the isolated antibody of claim
1.
12. A hybridoma that produces the isolated antibody of claim 1.
13. A hybridoma that produces the isolated antibody of claim 3.
14. An isolated antibody produced by immunizing an animal with a
protein whose sequence comprises of amino acid residues 21 to 116
of SEQ ID NO:238, wherein said antibody or portion thereof
specifically binds to the protein of SEQ ID NO:238.
15. The isolated antibody of claim 14, wherein said protein
specifically bound by said antibody is glycosylated.
16. The isolated antibody of claim 14 which is a monoclonal
antibody.
17. The isolated antibody of claim 14 which is a polyclonal
antibody.
18. The isolated antibody of claim 14 which is labeled.
19. The isolated antibody of claim 18, wherein the label is
selected from the group consisting of: (a) an enzyme label; (b) a
radioisotope; and (c) a fluorescent label.
20. An isolated cell that produces the isolated antibody of claim
14.
21. A hybridoma that produces the isolated antibody of claim
14.
22. A hybridoma that produces the isolated antibody of claim
16.
23. An isolated antibody or portion thereof that specifically binds
to a protein whose sequence consists of amino acid residues 1 to
116 of SEQ ID NO:238.
24. The isolated antibody or portion thereof of claim 23 wherein
said protein specifically bound by said antibody or portion thereof
is glycosylated.
25. The isolated antibody or portion thereof of claim 23 which is a
monoclonal antibody.
26. The isolated antibody or portion thereof of claim 23 which is a
polyclonal antibody.
27. The isolated antibody or portion thereof of claim 23 which is a
chimeric antibody.
28. The isolated antibody or portion thereof of claim 23 which is a
humanized antibody.
29. The isolated antibody or portion thereof of claim 23 which is a
human antibody.
30. The isolated antibody or portion thereof of claim 23 which is a
Fab fragment.
31. The isolated antibody or portion thereof of claim 23 which is
labeled.
32. The isolated antibody of claim 31, wherein the label is
selected from the group consisting of: (a) an enzyme label; (b) a
radioisotope; and (c) a fluorescent label.
33. An isolated cell that produces the isolated antibody of claim
23.
34. A hybridoma that produces the isolated antibody of claim
23.
35. A hybridoma that produces the isolated antibody of claim
25.
36. An isolated antibody or portion thereof that specifically binds
to a protein whose sequence consists of the amino acid sequence of
the secreted polypeptide encoded by the HLHFPO3 cDNA contained in
ATCC Deposit No. 209126.
37. The isolated antibody or portion thereof of claim 36, wherein
said protein specifically bound by said antibody or portion thereof
is glycosylated.
38. The isolated antibody or portion thereof of claim 36 which is a
monoclonal antibody.
39. The isolated antibody or portion thereof of claim 36 which is a
polyclonal antibody.
40. The isolated antibody or portion thereof of claim 36 which is a
chimeric antibody.
41. The isolated antibody or portion thereof of claim 36 which is a
humanized antibody.
42. The isolated antibody or portion thereof of claim 36 which is a
human antibody.
43. The isolated antibody or portion thereof of claim 36 which is a
Fab fragment.
44. The isolated antibody or portion thereof of claim 36 which is
labeled.
45. The isolated antibody of claim 44, where in the label is
selected from the group consisting of: (a) an enzyme label; (b) a
radioisotope; and (c) a fluorescent label.
46. An isolated cell that produces the isolated antibody of claim
36.
47. A hybridoma that produces the isolated antibody of claim
36.
48. A hybridoma that produces the isolated antibody of claim
38.
49. An isolated antibody produced by immunizing an animal with a
protein whose sequence comprises the amino acid sequence of the
secreted polypeptide encoded by the HLIFP03 cDNA contained in ATCC
Deposit No. 209126, wherein said antibody or portion thereof
specifically binds to the polypeptide encoded by the HLHFP03 cDNA
contained in ATCC Deposit No. 209126.
50. The isolated antibody of claim 49, wherein said protein
specifically bound by said antibody is glycosylated.
51. The isolated antibody of claim 49 which is a monoclonal
antibody.
52. The isolated antibody of claim 49 which is a polyclonal
antibody.
53. The isolated antibody of claim 49 which is labeled.
54. The isolated antibody of claim 53, wherein the label is
selected from the group consisting of: (a) an enzyme label; (b) a
radioisotope; and (c) a fluorescent label.
55. An isolated cell that produces the isolated antibody of claim
49.
56. A hybridoma that produces the isolated antibody of claim
49.
57. A hybridoma that produces the isolated antibody of claim
51.
58. An isolated antibody or portion thereof that specifically binds
to a protein whose sequence consists of the amino acid sequence of
the full-length polypeptide encoded by the HLHFP03 cDNA contained
in ATCC Deposit No. 209126;
59. The isolated antibody or portion thereof of claim 58 wherein
said protein specifically bound by said antibody or portion thereof
is glycosylated.
60. The isolated antibody or portion thereof of claim 58 which is a
monoclonal antibody.
61. The isolated antibody or portion thereof of claim 58 which is a
polyclonal antibody.
62. The isolated antibody or portion thereof of claim 58 which is a
chimeric antibody.
63. The isolated antibody or portion thereof of claim 58 which is a
humanized antibody.
64. The isolated antibody or portion thereof of claim 58 which is a
human antibody.
65. The isolated antibody or portion thereof of claim 58 which is a
Fab fragment.
66. The isolated antibody or portion thereof of claim 58 which is
labeled.
67. The isolated antibody of claim 66 wherein the label is selected
from the group consisting of: (a) an enzyme label; (b) a
radioisotope; and (c) a fluorescent label.
68. An isolated cell that produces the isolated antibody of claim
58.
69. A hybridoma that produces the isolated antibody of claim
58.
70. A hybridoma that produces the isolated antibody of claim 60.
Description
FIELD OF THE INVENTION
[0001] This invention relates to newly identified polynucleotides
and the polypeptides encoded by these polynucleotides, uses of such
polynucleotides and polypeptides, and their production.
BACKGROUND OF THE INVENTION
[0002] Unlike bacterium, which exist as a single compartment
surrounded by a membrane, human cells and other eucaryotes are
subdivided by membranes into many functionally distinct
compartments. Each membrane-bounded compartment, or organelle,
contains different proteins essential for the function of the
organelle. The cell uses "sorting signals," which are amino acid
motifs located within the protein, to target proteins to particular
cellular organelles.
[0003] One type of sorting signal, called a signal sequence, a
signal peptide, or a leader sequence, directs a class of proteins
to an organelle called the endoplasmic reticulum (ER). The ER
separates the membrane-bounded proteins from all other types of
proteins. Once localized to the ER, both groups of proteins can be
further directed to another organelle called the Golgi apparatus.
Here, the Golgi distributes the proteins to vesicles, including
secretory vesicles, the cell membrane, lysosomes, and the other
organelles.
[0004] Proteins targeted to the ER by a signal sequence can be
released into the extracellular space as a secreted protein. For
example, vesicles containing secreted proteins can fuse with the
cell membrane and release their contents into the extracellular
space--a process called exocytosis. Exocytosis can occur
constitutively or after receipt of a triggering signal. In the
latter case, the proteins are stored in secretory vesicles (or
secretory granules) until exocytosis is triggered. Similarly,
proteins residing on the cell membrane can also be secreted into
the extracellular space by proteolytic cleavage of a "linker"
holding the protein to the membrane.
[0005] Despite the great progress made in recent years, only a
small number of genes encoding human secreted proteins have been
identified. These secreted proteins include the commercially
valuable human insulin, interferon, Factor VIII, human growth
hormone, tissue plasminogen activator, and erythropoeitin. Thus, in
light of the pervasive role of secreted proteins in human
physiology, a need exists for identifying and characterizing novel
human secreted proteins and the genes that encode them. This
knowledge will allow one to detect, to treat, and to prevent
medical disorders by using secreted proteins or the genes that
encode them.
SUMMARY OF THE INVENTION
[0006] The present invention relates to novel polynucleotides and
the encoded polypeptides. Moreover, the present invention relates
to vectors, host cells, antibodies, and recombinant methods for
producing the polypeptides and polynucleotides. Also provided are
diagnostic methods for detecting disorders related to the
polypeptides, and therapeutic methods for treating such disorders.
The invention further relates to screening methods for identifying
binding partners of the polypeptides.
DETAILED DESCRIPTION
[0007] Definitions
[0008] The following definitions are provided to facilitate
understanding of certain terms used throughout this
specification.
[0009] In the present invention, "isolated" refers to material
removed from its original environment (e.g., the natural
environment if it is naturally occurring), and thus is altered "by
the hand of man" from its natural state. For example, an isolated
polynucleotide could be part of a vector or a composition of
matter, or could be contained within a cell, and still -be
"isolated" because that vector, composition of matter, or
particular cell is not the original environment of the
polynucleotide.
[0010] In the present invention, a "secreted" protein refers to
those proteins capable of being directed to the ER, secretory
vesicles, or the extracellular space as a result of a signal
sequence, as well as those proteins released into the extracellular
space without necessarily containing a signal sequence. If the
secreted protein is released into the extracellular space, the
secreted protein can undergo extracellular processing to produce a
"mature" protein. Release into the extracellular space can occur by
many mechanisms, including exocytosis and proteolytic cleavage.
[0011] As used herein, a "polynucleotide" refers to a molecule
having a nucleic acid sequence contained in SEQ ID NO:X or the cDNA
contained within the clone deposited with the ATCC. For example,
the polynucleotide can contain the nucleotide sequence of the full
length cDNA sequence, including the 5' and 3' untranslated
sequences, the coding region, with or without the signal sequence,
the secreted protein coding region, as well as fragments, epitopes,
domains, and variants of the nucleic acid sequence. Moreover, as
used herein, a "polypeptide" refers to a molecule having the
translated amino acid sequence generated from the polynucleotide as
broadly defined.
[0012] In the present invention, the full length sequence
identified as SEQ ID NO:X was often generated by overlapping
sequences contained in multiple clones (contig analysis). A
representative clone containing all or most of the sequence for SEQ
ID NO:X was deposited with the American Type Culture Collection
("ATCC"). As shown in Table 1, each clone is identified by a cDNA
Clone ID (Identifier) and the ATCC Deposit Number. The ATCC is
located at 10801 University Boulevard, Manassas, Va. 20110-2209,
USA. The ATCC deposit was made pursuant to the terms of the
Budapest Treaty on the international recognition of the deposit of
microorganisms for purposes of patent procedure.
[0013] A "polynucleotide" of the present invention also includes
those polynucleotides capable of hybridizing, under stringent
hybridization conditions, to sequences contained in SEQ ID NO:X,
the complement thereof, or the cDNA within the clone deposited with
the ATCC. "Stringent hybridization conditions" refers to an
overnight incubation at 42.degree. C. in a solution comprising 50%
formamide, 5.times. SSC (750 mM NaCl, 75 mM sodium citrate), 50 mM
sodium phosphate (pH 7.6), 5.times. Denhardt's solution, 10%
dextran sulfate, and 20 .mu.g/ml denatured, sheared salmon sperm
DNA, followed by washing the filters in 0.1.times. SSC at about
65.degree. C.
[0014] Also contemplated are nucleic acid molecules that hybridize
to the polynucleotides of the present invention at lower stringency
hybridization conditions. Changes in the stringency of
hybridization and signal detection are primarily accomplished
through the manipulation of formamide concentration (lower
percentages of formamide result in lowered stringency); salt
conditions, or temperature. For example, lower stringency
conditions include an overnight incubation at 37.degree. C. in a
solution comprising 6.times. SSPE (20.times. SSPE=3M NaCl; 0.2M
NaH.sub.2PO.sub.4; 0.02M EDTA, pH 7.4), 0.5% SDS, 30% formamide,
100 ug/ml salmon sperm blocking DNA; followed by washes at
50.degree. C. with 1.times. SSPE, 0.1% SDS. In addition, to achieve
even lower stringency, washes performed following stringent
hybridization can be done at higher salt concentrations (e.g.
5.times. SSC).
[0015] Note that variations in the above conditions may be
accomplished through the inclusion and/or substitution of alternate
blocking reagents used to suppress background in hybridization
experiments. Typical blocking reagents include Denhardt's reagent,
BLOTTO, heparin, denatured salmon sperm DNA, and commercially
available proprietary formulations. The inclusion of specific
blocking reagents may require modification of the hybridization
conditions described above, due to problems with compatibility.
[0016] Of course, a polynucleotide which hybridizes only to polyA+
sequences (such as any 3' terminal polyA+ tract of a cDNA shown in
the sequence listing), or to a complementary stretch of T (or U)
residues, would not be included in the definition of
"polynucleotide," since such a polynucleotide would hybridize to
any nucleic acid molecule containing a poly (A) stretch or the
complement thereof (e.g., practically any double-stranded cDNA
clone).
[0017] The polynucleotide of the present invention can be composed
of any polyribonucleotide or polydeoxribonucleotide, which may be
unmodified RNA or DNA or modified RNA or DNA. For example,
polynucleotides can be composed of single- and double-stranded DNA,
DNA that is a mixture of single- and double-stranded regions,
single- and double-stranded RNA, and RNA that is mixture of single-
and double-stranded regions, hybrid molecules comprising DNA and
RNA that may be single-stranded or, more typically, double-stranded
or a mixture of single- and double-stranded regions. In addition,
the polynucleotide can be composed of triple-stranded regions
comprising RNA or DNA or both RNA and DNA. A polynucleotide may
also contain one or more modified bases or DNA or RNA backbones
modified for stability or for other reasons. "Modified" bases
include, for example, tritylated bases and unusual bases such as
inosine. A variety of modifications can be made to DNA and RNA;
thus, "polynucleotide" embraces chemically, enzymatically, or
metabolically modified forms.
[0018] The polypeptide of the present invention can be composed of
amino acids joined to each other by peptide bonds or modified
peptide bonds, i.e., peptide isosteres, and may contain amino acids
other than the 20 gene-encoded amino acids. The polypeptides may be
modified by either natural processes, such as posttranslational
processing, or by chemical modification techniques which are well
known in the art. Such modifications are well described in basic
texts and in more detailed monographs, as well as in a voluminous
research literature. Modifications can occur anywhere in a
polypeptide, including the peptide backbone, the amino acid
side-chains and the amino or carboxyl termini. It will be
appreciated that the same type of modification may be present in
the same or varying degrees at several sites in a given
polypeptide. Also, a given polypeptide may contain many types of
modifications. Polypeptides may be branched , for example, as a
result of ubiquitination, and they may be cyclic, with or without
branching. Cyclic, branched, and branched cyclic polypeptides may
result from posttranslation natural processes or may be made by
synthetic methods. Modifications include acetylation, acylation,
ADP-ribosylation, amidation, covalent attachment of flavin,
covalent attachment of a heme moiety, covalent attachment of a
nucleotide or nucleotide derivative, covalent attachment of a lipid
or lipid derivative, covalent attachment of phosphotidylinositol,
cross-linking, cyclization, disulfide bond formation,
demethylation, formation of covalent cross-links, formation of
cysteine, formation of pyroglutamate, formylation,
gamma-carboxylation, glycosylation, GPI anchor formation,
hydroxylation, iodination, methylation, myristoylation, oxidation,
pegylation, proteolytic processing, phosphorylation, prenylation,
racemization, selenoylation, sulfation, transfer-RNA mediated
addition of amino acids to proteins such as arginylation, and
ubiquitination. (See, for instance, PROTEINS--STRUCTURE AND
MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and
Company, New York (1993); POSTTRANSLATIONAL COVALENT MODIFICATION
OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, pgs.
1-12 (1983); Seifter et al., Meth Enzymol 182:626-646 (1990);
Rattan et al., Ann NY Acad Sci 663:48-62 (1992).)
[0019] "SEQ ID NO:X" refers to a polynucleotide sequence while "SEQ
ID NO:Y" refers to a polypeptide sequence, both sequences
identified by an integer specified in Table 1.
[0020] "A polypeptide having biological activity" refers to
polypeptides exhibiting activity similar, but not necessarily
identical to, an activity of a polypeptide of the present
invention, including mature forms, as measured in a particular
biological assay, with or without dose dependency. In the case
where dose dependency does exist, it need not be identical to that
of the polypeptide, but rather substantially similar to the
dose-dependence in a given activity as compared to the polypeptide
of the present invention (i.e., the candidate polypeptide will
exhibit greater activity or not more than about 25-fold less and,
preferably, not more than about tenfold less activity, and most
preferably, not more than about three-fold less activity relative
to the polypeptide of the present invention.)
[0021] Polynucleotides and Polypeptides of the Invention
[0022] Features of Protein Encoded by Gene No: 1
[0023] This gene is expressed primarily in cerebellum. Therefore,
polynucleotides and polypeptides of the invention are useful as
reagents for differential identification of the tissue(s) or cell
type(s) present in a biological sample and for diagnosis of
diseases and conditions which include, but are not limited to,
neural disorders, particularly damage to the cerebellum or
additional CNS tissues caused by injuries, which include, but are
not limited to, trauma or ischemia. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the central nervous system (CNS),
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
neural, and cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0024] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 150 as residues: Pro-7 to Cys-21, Leu-25 to
Ser-30.
[0025] The tissue distribution in cerebellum indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection, treatment, and/or prevention of
neurodegenerative disease states, behavioral disorders, or
inflammatory conditions which include, but are not limited to
Alzheimer's Disease, Parkinson's Disease, Huntington's Disease,
Tourette Syndrome, meningitis, encephalitis, demyelinating
diseases, peripheral neuropathies, neoplasia, trauma, congenital
malformations, spinal cord injuries, ischemia and infarction,
aneurysms, hemorrhages, schizophrenia, mania, dementia, paranoia,
obsessive compulsive disorder, depression, panic disorder, learning
disabilities, ALS, psychoses, autism, and altered behaviors,
including disorders in feeding, sleep patterns, balance, and
perception.
[0026] In addition, elevated expression of this gene product in
regions of the brain indicates it plays a role in normal neural
function. Potentially, this gene product is involved in synapse
formation, neurotransmission, learning, cognition, homeostasis, or
neuronal differentiation or survival. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed tissues.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tumors and tissues.
[0027] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:11 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1868 of SEQ ID NO:11, b is an integer
of 15 to 1882, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:11, and where b is greater
than or equal to a+14.
[0028] Features of Protein Encoded by Gene No: 2
[0029] The translation product of this gene was found to have
homology to the human env endogenous retrovirus protein (See
Genbank Accession No, gil757872), which is thought to play a
contributing role in the events leading up to the onset of cancer
or of proliferative disorders in teratocarcinoma cell lines. In
specific embodiments, polypeptides of the invention comprise the
following amino acid sequence:
1 VDPRVRRFWEDPEYPPVAVMSRLMLRRIPTVMSNTHRTQPSTWEQIKKLSQMVGENLRKAGQPVT
(SEQ ID NO: 289), VRRFWEDPEYPPVAVMSRLMLRRIP (SEQ ID NO: 290),
SNTHRTQPSTWEQIKKLSQMVGENLRK (SEQ ID NO: 291),
SACHSHTVFNWSEQNGQMVQMVRRMARVPIIWNHGSIGAPQPQMIWPIVGA (SEQ ID NO:
292), KHKDLWQLLIALNKIKIWERIKKHLEGHSANLSLDIAKYIYIFKASQAHLT
LMPELECSKELQTD MARVPIIWNHGSIGAPQPQMIWPIV (SEQ ID NO: 293),
RIKKHLEGHSANLSL DIAKYIYIFKASQAHLT (SEQ ID NO: 294),
VFLQQGLTQRSVILIGHICQFWLAIMPGYNHFMTQLHMLSGLNI- YH (SEQ ID NO: 295),
NKSAPIIEAYHP QKSICKQN IGHICQFWLAIMPGYNHFMTQLHMLSGL (SEQ ID NO:
296), SIPGTPDLNARTGVLEGAADRLAASNPLKWIKTLRSSVISMMIVLLICVVCLYIVCRC
(SEQ ID NO: 297), VLEGAADRLAASNPLKWIKTLRSSVIS (SEQ ID NO: 298),
LTVTKLPWLFIALQNKRMGTSWEQA (SEQ ID NO: 299), and/or
PKSGHKLAPKLVINKISAALSHACDSLTPTLEGCRFTGM RARNNWPTQGG
MGTSWEQAPKSGHKLAPKLVINKISAALS (SEQ ID NO: 300).
[0030] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0031] This gene is expressed primarily in PHA stimulated
T-cells.
[0032] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of the following diseases and conditions which include,
but are not limited to, immune disorders, particularly autoimmune,
inflammatory, or immunodeficiency diseases, in addition to,
proliferative conditions such as cancers. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
-the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
immune, hematopoietic, teratocarcinoma, and cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum; plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0033] The tissue distribution in T-cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. The expression of this gene product
indicates a role in the regulation of the proliferation; survival;
differentiation; and/or activation of potentially all hematopoietic
cell lineages, including blood stem cells. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer e.g. by boosting immune responses. Since
the gene is expressed in cells of lymphoid origin, the natural gene
product may be involved in immune functions. Therefore it may be
also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS, and
leukemia.
[0034] In addition, this gene product may have commercial utility
in the expansion of stem cells and committed progenitors of various
blood lineages, and in the differentiation and/or proliferation of
various cell types. The protein may also show utility in the
development of novel inhibitors to viral infections, or the protein
may be useful in the development of novel vectors, such as those
used in gene therapy, and/or immuno-therapy which could lead to the
amelioration of disease of disease states. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed tumors and
tissues.
[0035] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:12 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1576 of SEQ ID NO:12, b is an integer
of 15 to 1590, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:12, and where b is greater
than or equal to a+14.
[0036] Features of Protein Encoded by Gene No: 3
[0037] The translation product of this gene was shown to have
homology to the human retrovirus-related reverse transcriptase
pseudogene (See Genbank Accession No.
pirlA25313.vertline.GNHULL).In specific embodiments, polypeptides
of the invention comprise the following amino acid sequence:
2 STHASVQKKDLTKFSAHSWLKKKKTFRKMIMEEIFLNLIKNIYKSPYSQCNT (SEQ ID NO:
301), VRSEKGFDKIQCPFMVK (SEQ ID NO: 302),
FSKPSSYKTYIPKINLHFYILLMNIWETIKIVPLNNCFTKMNYLGI (SEQ ID NO: 303),
KKETKLSLFANDMI (SEQ ID NO: 304), and/or SPLLFNILLEVLSSAVRKEKELK
(SEQ ID NO: 305).
[0038] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0039] This gene is expressed primarily in PHA activated T
cells.
[0040] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions: immune or hematopoietic
disorders, particularly inflammation, immunodeficiencies, and
autoimmune diseases. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune,
hematopoietic, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0041] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 152 as residues: Ile-14 to Thr-24.
[0042] The tissue distribution in T-cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. The expression of this gene product
indicates a role in the regulation of the proliferation; survival;
differentiation; and/or activation of potentially all hematopoietic
cell lineages, including blood stem cells. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer e.g. by boosting immune responses. Since
the gene is expressed in cells of lymphoid origin, the natural gene
product may be involved in immune functions. Therefore it may be
also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS, and
leukemia.
[0043] In addition, this gene product may have commercial utility
in the expansion of stem cells and committed progenitors of various
blood lineages, and in the differentiation and/or proliferation of
various cell types. Alternatively, the homology to a reverse
transcriptase human gene may implicate this gene as providing
utility in the understanding of host-viral interactions,
particularly those involving retroviruses and other
integration-dependent viruses. Moreover, the protein may show
utility in the development of novel inhibitors to viral infection,
and thus to the amelioration of human diseases and conditions.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0044] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:13 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1359 of SEQ ID NO:13, b is an integer
of 15 to 1373, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:13, and where b is greater
than or equal to a+14.
[0045] Features of Protein Encoded by Gene No: 4
[0046] The translation product of this gene shares sequence
homology with npdcf-1 which is thought to be important in promoting
the survival of bi-potential glial progenitor cells (See Genbank
Accession No. gi.vertline.456107). One embodiment of this gene
comprises polypeptides of the following amino acid sequence:
3 LRRPSTPLRRPWLHLQLPRISLGDQRLAQSAEMYHYQHQRQQMLSLERHKEPP (SEQ ID NO:
306), KELDTALRMRRMRTETSRCTSARAWPRPGKWRCATICSTTPHCPRPCRPP
AHRLHCHDLEADRRPLAPR RATQGAGHGSSDEENEDG (SEQ ID NO: 307),
DFTVYECPGMAPTGEMEVRNHLFD HAALSAPLPAPSSPLALP
KAEYATAKALATPAATPDLAWGPAPGTERGDVPLPAPT- ATDVVPGAA (SEQ ID NO: 308),
SAEMYHYQHQRQQML (SEQ ID NO: 309), LERHKEPPKEL (SEQ ID NO: 310),
AKCPPGAHACGP (SEQ ID NO: 311), PVHMSPLEP (SEQ ID NO: 312),
WCRLQREIRLTQ (SEQ ID NO: 313), SSDEENEDGDFTVYECPG (SEQ ID NO: 314),
APTGEMEVRN (SEQ ID NO: 315), CPGSLDCALK (SEQ ID NO: 316),
RATQGAGHGSSDEENEDGDETVYECPGMAPTGEMEVRNHLFDHAALS (SEQ ID NO: 317),
APLPAPSSPLALP NEDGDFTVYECPGMAPTGEMEV (SEQ ID NO: 318),
RPTRPSSSCVLPRCLRCSRRGARSPRRAPGLAVPCCPGGGAEGWR (SEQ ID NO: 319),
RRCLRPPRGTCGCCGCCSPASSSAPPCVEPPPATRNVAACPGSL- DCALKKRA
SVLLVHMPVGLPSALPXGTAKACFAXMRRASXGGRAQPXLEMRLIPGPR ELARKGIWTSIPP
RCLRCSRRGARSPRRAPGLAVPCCP (SEQ ID NO: 320), and/or
GSLDCALKKRASVLLVHMPVGLPSALPXGTAK- AC (SEQ ID NO: 321).
[0047] Additional embodiments is the polynucleotides encoding these
polypeptides.
[0048] This gene is expressed primarily in cerebellum, synovial
sarcoma, and to a lesser extent, in several other cancer cell
lines.
[0049] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neural or skeletal disorders, particularly tumors
characterized by cells of a relatively undifferentiated state,
including neural tumors. Similarly, polypeptides and antibodies
directed to those polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the synovial fluid, prostate,
breast and uterus, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types (e.g., neural, skeletal, and cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0050] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 153 as residues: Pro-6 to Arg-11, Glu-52 to
Gly-59.
[0051] The tissue distribution in the cerebellum, combined with the
homology to the human npdcf-1 protein indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for diagnosing and treating tumors that contain relatively
high numbers of undifferentiated cells. Moreover, this gene is
useful for the detection, treatment, and/or prevention of
neurodegenerative disease states and behavioural disorders such as
Alzheimers Disease, Parkinsons Disease, Huntingtons Disease,
Tourette Syndrome, schizophrenia, mania, dementia, paranoia,
obsessive compulsive disorder, panic disorder, learning
disabilities, ALS, psychoses , autism, and altered behaviors,
including disorders in feeding, sleep patterns, balance, and
perception.
[0052] In addition, the gene or gene product may also play a role
in the treatment and/or detection of developmental disorders
associated with the developing embryo, sexually-linked disorders,
or disorders of the cardiovascular system. Alternatively, the
expression of this gene product in synovium would suggest a role in
the detection and treatment of disorders and conditions affecting
the skeletal system, in particular osteoporosis, bone cancer, as
well as, disorders afflicting connective tissues (e.g., arthritis,
trauma, tendonitis, chrondomalacia and inflammation), such as in
the diagnosis or treatment of various autoimmune disorders such as
rheumatoid arthritis, lupus, scleroderma, and dermatomyositis as
well as dwarfism, spinal deformation, and specific joint
abnormalities as well as chondrodysplasias (i.e. spondyloepiphyseal
dysplasia congenita, familial osteoarthritis, Atelosteogenesis type
II, metaphyseal chondrodysplasia type Schmid).
[0053] Moreover, the protein may be useful for inducing astroglial
proliferation and promoting neuronal survival, in addition to other
highly vascular tissues. The protein can also be used to regulate
cellular metabolism (e.g., through the modulation of protein
expression). Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0054] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:14 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1128 of SEQ ID NO:14, b is an integer
of 15 to 1142, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:14, and where b is greater
than or equal to a+14.
[0055] Features of Protein Encoded by Gene No: 5
[0056] The translation product of this gene was found to have
homology to the RoBo-1 protein from Rattus norvegicus (See Genbank
Accession No.gi.vertline.2895563 (AF041083)) which is thought to be
important as a mediator in bone remodeling. In specific
embodiments, polypeptides of the invention comprise the following
amino acid sequence: DSHQARSRRLEALWSPSLGEVSSST (SEQ ID NO: ).
Polynucleotides encoding these polypeptides are also encompassed by
the invention.
[0057] This gene is expressed primarily in colon, pituitary, and to
a lesser extent in fetal lung and fibrosarcoma.
[0058] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, endocrine, gastrointestinal, pulmonary, skeletal, or
developmental and proliferative disorders, particularly those
effecting the Gut/pituitary/hypothalamic axis. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
digestive system and regulation of feeding, expression of this gene
at significantly higher or lower levels may be routinely detected
in certain tissues or cell types (e.g., gastrointesinal, endocrine,
developmental, skeletal, pulmonary, and cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, amniotic
fluid, pulmonary surfactant or sputum, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0059] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 154 as residues: Asn-26 to Cys-32, Cys-100 to
Leu-112, Cys-128 to Ser-135.
[0060] The tissue distribution in colon and pituitary indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for treating disorders related to the intake and
utilization of food since this gene is expressed in the digestive
tract and a CNS site involved in regulation of weight homeostasis.
The secreted protein can also be used to determine biological
activity, to raise antibodies, as tissue markers, to isolate
cognate ligands or receptors, to identify agents that modulate
their interactions, and as nutritional supplements. It may also
have a very wide range of biological activities. Typical of these
are cytokine, cell proliferation/differentiation modulating
activity or induction of other cytokines;
immunostimulating/immunosuppres- sant activities (e.g., for
treating human immunodeficiency virus infection, cancer, autoimmune
diseases and allergy); regulation of hematopoiesis (e.g., for
treating anemia or as adjunct to chemotherapy); stimulation or
growth of bone, cartilage, tendons, ligaments and/or nerves (e.g.,
for treating wounds, stimulation of follicle stimulating hormone
(for control of fertility); chemotactic and chemokinetic activities
(e.g., for treating infections, tumors); hemostatic or thrombolytic
activity (e.g., for treating hemophilia, cardiac infarction etc.);
anti-inflammatory activity (e.g., for treating septic shock,
Crohn's disease); as antimicrobials; for treating psoriasis or
other hyperproliferative diseases; for regulation of metabolism,
and behavior. Also contemplated is the use of the corresponding
nucleic acid in gene therapy procedures. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0061] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:15 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1020 of SEQ ID NO:15, b is an integer
of 15 to 1034, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO: 15, and where b is greater
than or equal to a+14.
[0062] Features of Protein Encoded by Gene No: 6
[0063] The translation product of this gene shares sequence
homology with Cortical granule lectin which is thought to be
important in blocking polyspermy (See Genbank Accession No.
gnl.vertline.PID.vertline.e1181610)- . In specific embodiments,
polypeptides of the invention comprise the following amino acid
sequence:
4 RSCKEIKD (SEQ ID NO: 323), GGGWTLVASVHEN (SEQ ID NO: 324),
ADYPEGDGNWANYNTFGSA (SEQ ID NO: 325), ATSDDYKNPGYYDI (SEQ ID NO:
326), CIGGGGYFPEA (SEQ ID NO: 327), DSDKIT (SEQ ID NO: 329),
YQTFCDMT (SEQ ID NO: 330), and/or EITEAAVLLFY (SEQ ID NO: 328).
[0064] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0065] This gene is expressed primarily in benign and metastatic
colon, and to a lesser extent in HEL cells.
[0066] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, gastrointestinal, reproductive, or developmental
disorders, particularly cancer, or inflammatory conditions.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the digestive system, expression of this gene at significantly
higher or lower levels may be detected in certain tissues or cell
types (e.g., gastrointestinal, reproductive, proliferating, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, seminal fluid, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0067] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 155 as residues: Arg-15 to Ser-33, Pro-35 to
Cys-41.
[0068] The tissue distribution in colon, combined with the homology
to cortical granule lectins indicates that polynucleotides and
polypeptides corresponding to this gene are useful for treating
disorders of the colon. These may include diseases related to
damage or chronic inflammation as well as tumors of the colon. The
product may also be useful for the identification of colon cancer
metastasis and, as a secreted protein, may have diagnostic and
prognostic applications. Moreover, the protein is useful in the
treatment, detection, and/or prevention of reproductive disorders,
particularly normal testicular function, in addition having utility
in the development of contraceptives, or in the treatment of
polyspermy. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tumors and tissues.
[0069] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:16 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1184 of SEQ ID NO:16, b is an integer
of 15 to 1198, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:16, and where b is greater
than or equal to a+14.
[0070] Features of Protein Encoded by Gene No: 7
[0071] This gene is expressed primarily in eight week human
embryos.
[0072] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, fetal and/or developmental abnormalities. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
developing fetus, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types (e.g., developing, differentiating, and cancerous and
wounded tissues) or bodily fluids (e.g., amniotic fluid, serum,
plasma, lymph, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0073] The tissue distribution in eight week old tissue indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for detecting embryonic abnormalities, in particular
congenital abnormalities, which include, but are not limited to
Tay-Sachs disease, phenylkenonuria, galactosemia, hyperlipidemias,
porphyrias, and Hurler's syndrome. Expression within embryonic
tissue and other cellular sources marked by proliferating cells
indicates that this protein may play a role in the regulation of
cellular division, and may show utility in the diagnosis,
treatment, and/or prevention of cancer and other proliferative'
disorders. Similarly, developmental tissues rely on decisions
involving cell differentiation and/or apoptosis in pattern
formation. Thus this protein may also be involved in apoptosis or
tissue differentiation and could again be useful in cancer therapy.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0074] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:17 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1433 of SEQ ID NO:17, b is an integer
of 15 to 1447, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:17, and where b is greater
than or equal to a+14.
[0075] Features of Protein Encoded by Gene No: 8
[0076] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
5 MGKRAHEVRRPPHSRPLHGTPAGWVLDPSGYKDVTQDA (SEQ ID NO: 331),
EVMEVLQNLYRTKSFLFVGCGETLRDQIFQALFLYSVPNKVDLEHYMLVLKE
NEDHFFKHQADMLLHGIKVVSYGDCFDHFPGYVQDLATQICKQQSPGHLYSN SWSATPDGRGGP
VLDPSGYKDVTQDAEVMEVLQNLYRT (SEQ ID NO: 332), YSVPNKVDLEHY
MLVLKENEDHFFKII (SEQ ID NO: 333), DLATQICKQQSPGHLYSNSWSATPD (SEQ ID
NO: 334), RRMKTISLSIRQICFC (SEQ ID NO: 335),
TESKLYPTGTVLTTFQDMCKTLPLRSANSKAQDICTRIHGVPLLMGEEAHDSD
SHASDRGHHTMLPLPAGSFSESSHQAWEVEMLIAWTAPHYWVMHARTVQRGS
TESKLYPTGTVLTTFQDMCKTLPLRSA (SEQ ID NO: 336), and/or LMGEEAH
DSDSHASDRGHHTMLPLPAG (SEQ ID NO: 337).
[0077] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0078] This gene is expressed primarily in endothelial cells, and
to a lesser extent in lymph node, tonsils, heart and spinal
cord.
[0079] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, vascular diseases, such as restenosis, including
disorders of the integumentary system. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the vascular system, expression
of this gene at significantly higher or lower levels may be
routinely detected in certain tissues or cell types (e.g.,
vascular, integumentary, immune, hematopoietic, neural,
cardiovascular, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0080] The tissue distribution in endothelial cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for treating diseases of the vasculature including problems
associated with diabetes and restenosis following angioplasty.
Moreover, the protein is useful in the detection, treatment, and/or
prevention of a variety of other vascular conditions, which
include, but are not limited to, stroke, microvascular disease,
aneurysm, vascular leak syndrome, or embolism. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0081] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:18 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1408 of SEQ ID NO:18, b is an integer
of 15 to 1422, where both a and b correspond to the-positions of
nucleotide residues shown in SEQ ID NO:18, and where b is greater
than or equal to a+14.
[0082] Features of Protein Encoded by Gene No: 9
[0083] The translation product of this gene was shown to have
homology to the Gcap1 gene product of Mus musculus, which is
specifically expressed in cerebellum and appears to be
developmentally regulated (See Genbank Accession No.
gi.vertline.862343). In specific embodiments, polypeptides of the
invention comprise the following amino acid sequence:
6 LCAVEKTRTFTRGDCGPNRHHKHVLKAKDNNHIQRHQFSSTLEFS (SEQ ID NO:338),
SNSTDGLKYICVYLYVCTHPCIYIYLSAHTLHMYTHYLCKI SST
LEFSSNSTDGLKYICVYLYVCTHPCIY (SEQ ID NO:339), STSVCICTCAHTHVYI (SEQ
ID NO:340), FIYLHTHYICIHTIYVKYNICIM- HINSNKCICVIFKIEQLYLEVVNAENWFYC
IHTIYVKYNICIMHIN SNKCICVITFKIEQLY (SEQ ID NO:341), and/or NSAVTVQMA
(SEQ ID NO:342).
[0084] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0085] This gene is expressed primarily in fetal lung, endothelial
cells and to a lesser extent, in astrocytes and fetal brain.
[0086] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, vasdcular, developmental, neural, or proliferative
conditions, particularly endothelial cell proliferation, such as
occurs in restenosis. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the vascular system, expression
of this gene at significantly higher or lower levels may be
routinely detected in certain tissues or cell types (e.g.,
developmental, neural, vascular, and cancerous and wounded tissues)
or bodily fluids (e.g., amniotic fluid, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression, level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0087] The tissue distribution in fetal brain, in addition to the
homology to a brain-specific regulatory protein indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection, treatment, or prevention of
neurodegenerative disease states and behavioural disorders such as
Alzheimers Disease, Parkinsons Disease, Huntingtons Disease,
Tourette Syndrome, schizophrenia, mania, dementia, paranoia,
obsessive compulsive disorder, panic disorder, learning
disabilities, ALS, psychoses , autism; and altered behaviors,
including disorders in feeding, sleep patterns, balance, and
perception. In addition, the gene or gene product may also play a
role in the treatment and/or detection of developmental disorders
associated with the developing embryo, sexually-linked disorders,
or disorders of the cardiovascular system.
[0088] Alternatively, the tissue distribution indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for treating abnormal proliferation of endothelial cells
such as occurs upon injury to the lung or arteries. Moreover, this
protein may play a role in the regulation of cellular division, and
may show utility in the diagnosis and treatment of cancer and other
proliferative disorders. Similarly, developmental tissues rely on
decisions involving cell differentiation and/or apoptosis in
pattern formation. Thus this protein may also be involved in
apoptosis or tissue differentiation and could again be useful in
cancer therapy. Protein, as well as, antibodies directed against
the protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0089] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:19 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1093 of SEQ ID NO:19, b is an integer
of 15 to 1107, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:19, and where b is greater
than or equal to a+14.
[0090] Features of Protein Encoded by Gene No: 10
[0091] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: TKTSTPLR (SEQ ID NO:
343). Polynucleotides encoding these polypeptides are also
encompassed by the invention. The gene encoding the disclosed cDNA
is believed to reside on chromosome 12. Accordingly,
polynucleotides related to this invention are useful as a marker in
linkage analysis for chromosome 12.
[0092] This gene is expressed primarily in infant brain and fetal
tissues.
[0093] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, developmental abnormalities or neural disorders,
particularly gestational conditions, such as spina bifida.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., developing, neural,
differentiating, and cancerous and wounded tissues) or bodily
fluids (e.g., amniotic fluid, serum, lymph, plasma, urine, synovial
fluid and spinal fluid) or another tissue or cell sample taken from
an individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0094] The tissue distribution in infant brain and fetal tissues
suggests that the protein product of this clone is useful for the
detection/treatment of neurodegenerative disease states and
behavioural disorders such as Alzheimers Disease, Parkinsons
Disease, Huntingtons Disease, Tourette Syndrome, schizophrenia,
mania, dementia, paranoia, obsessive compulsive disorder, panic
disorder, learning disabilities, ALS, psychoses, autism, and
altered behaviors, including disorders in feeding, sleep patterns,
balance, and perception. In addition, the gene or gene product may
also play a role in the treatment and/or detection of developmental
disorders associated with the developing embryo, sexually-linked
disorders, or disorders of the cardiovascular system.
Alternatively, the tissue distribution suggests that the protein
product of this clone is useful for the diagnosis, treatment,
and/or prevention of cancer and other proliferative disorders
Moreover, the expression within fetal tissue and other cellular
sources marked by proliferating cells suggests that this protein
may play a role in the regulation of cellular division. Similarly,
embryonic development also involves decisions involving cell
differentiation and/or apoptosis in pattern formation. Thus this
protein may also be involved in apoptosis or tissue differentiation
and could again be useful in cancer therapy. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0095] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:20 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1169 of SEQ ID NO:20, b is an integer
of 15 to 1183, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:20, and where b is greater
than or equal to a+14.
[0096] Features of Protein Encoded by Gene No: 11
[0097] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: VCIPGAAGLSVLLG (SEQ ID
NO: 344). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0098] This gene is expressed primarily in fetal kidney.
[0099] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, urogenital, renal, or developmental disorders,
particularly renal failure, tumors of the kidney, and/or
developmental abnormalities associated with the kidney. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the renal
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., urological, renal, developmental, and cancerous and wounded
tissues) or bodily fluids (e.g., amniotic fluid, lymph, serum,
plasma, urine, synovial fluid and spinal fluid) or another tissue
or cell sample taken from an individual having such a disorder,
relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0100] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 160 as residues: Gln-26 to Gln-34.
[0101] The tissue distribution in fetal kidney indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of cancer
and other proliferative disorders, particularly renal disorders.
Expression within embryonic tissue and other cellular sources
marked by proliferating cells indicates that this protein may play
a role in the regulation of cellular division. Similarly, embryonic
development also involves decisions involving cell differentiation
and/or apoptosis in pattern formation. Thus this protein may also
be involved in apoptosis or tissue differentiation and could again
be useful in cancer therapy.
[0102] Moreover, the protein product of this gene could be used in
the treatment and/or detection of kidney diseases including
nephritus, renal tubular acidosis, proteinuria, pyuria, edema,
pyelonephritis, hydronephritis, nephrotic syndrome, crush syndrome,
glomerulonephritis, hematuria, renal colic and kidney stones, in
addition to Wilm's Tumor Disease, and congenital kidney
abnormalities such as horseshoe kidney, polycystic kidney, and
Falconi's syndrome. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0103] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:21 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1406 of SEQ ID NO:21, b is an integer
of 15 to 1420, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:21, and where b is greater
than or equal to a+14.
[0104] Features of Protein Encoded by Gene No: 12
[0105] The gene encoding the disclosed cDNA is believed to reside
on chromosome 17. Accordingly, polynucleotides related to this
invention are useful as a marker in linkage analysis for chromosome
17.
[0106] This gene is expressed primarily in breast, fetal kidney,
and T-cells.
[0107] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive, immune, developmental, or renal
disorders, particularly autoimmune diseases, chronic inflammatory
conditions, or urogenital disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune,
reproductive, cancerous and wounded tissues) or bodily fluids
(e.g., breast milk, lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0108] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 161 as residues: His-2 to Lys-7, Ser-28 to
Glu-35.
[0109] The tissue distribution in breast and T-cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Moreover, the expression of this gene
product indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of potentially all hematopoietic
cell lineages, including blood stem cells. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer e.g. by boosting immune responses.
[0110] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immune deficiency diseases such as
AIDS, and leukemia. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tumors and tissues. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types.
[0111] Alternatively, the expression within fetal tissue indicates
this protein may play a role in the regulation of cellular
division, and may show utility in the diagnosis and treatment of
cancer and other proliferative disorders. Similarly, developmental
tissues rely on decisions involving cell differentiation and/or
apoptosis in pattern formation. Thus this protein may also be
involved in apoptosis or tissue differentiation and could again be
useful in cancer therapy. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0112] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:22 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1561 of SEQ ID NO:22, b is an integer
of 15 to 1575, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:22, and where b is greater
than or equal to a+14.
[0113] Features of Protein Encoded by Gene No: 13
[0114] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
7 SILPVEMAAAVAGMLRGGLLPQAGRLPTLQTVRYGSKAVTRHRRV (SEQ ID NO: 345),
and/or AGMLRGGLLPQAGRLPTLQTVRYGSK (SEQ ID NO: 346).
[0115] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0116] This gene is expressed primarily in the frontal cortex of
the brain.
[0117] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neural disorders, particularly neurodegenerative
disorders, ischemia, Alzheimer's, or Parkinson's. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
central nervous system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., neural, and cancerous and wounded tissues) or
bodily fluids (e.g., serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0118] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 162 as residues: Glu-31 to Gly-37.
[0119] The tissue distribution in frontal cortex indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses ,
autism, and altered behaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing embryo.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tumors and tissues.
[0120] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:23 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 527 of SEQ ID NO:23, b is an integer
of 15 to 541, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:23, and where b is greater
than or equal to a+14.
[0121] Features of Protein Encoded by Gene No: 14
[0122] The gene encoding the disclosed cDNA is believed to reside
on chromosome 1. Accordingly, polynucleotides related to this
invention are useful as a marker in linkage analysis for chromosome
1.
[0123] This gene is expressed primarily in ovary, and to a lesser
extent, in infant brain.
[0124] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive, neural, or developmental disorders,
particularly cancers and other diseases of the reproductive system
including ovarian cysts and hormonal disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the female
reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., reproductive, neural, developmental, and
cancerous and wounded tissues) or bodily fluids (e.g., amniotic
fluid, lymph, seminal fluid, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0125] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 163 as residues: Ser-32 to Glu-37.
[0126] The tissue distribution in ovarian tissue indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for diagnosis and intervention of ovarian tumors, in
addition to other tumors where expression has been indicated.
Alternatively, expression within the fetal brain indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses ,
autism, and altered behaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing
embryo.
[0127] Moreover, the expression within fetal tissue indicates this
protein may play a role in the regulation of cellular division, and
may show utility in the diagnosis and treatment of cancer and other
proliferative disorders. Similarly, developmental tissues rely on
decisions involving cell differentiation and/or apoptosis in
pattern formation. Thus this protein may also be involved in
apoptosis or tissue differentiation and could again be useful in
cancer therapy. Protein, as well as, antibodies directed against
the protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0128] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:24 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 819 of SEQ ID NO:24, b is an integer
of 15 to 833, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:24, and where b is greater
than or equal to a+14.
[0129] Features of Protein Encoded by Gene No: 15
[0130] The translation product of this gene was shown to have
homology to the highly conserved ras gene which is known to be
important in the regulation of cell growth, and thus has been shown
to serve as an inducible oncogene in eukaryotic tissues (See
Genbank Accession No. gb.vertline.Z11804.vertline.DDRASX). When
tested against PC12 (rat pheochromocytoma cells) and NIH3T3 cell
lines, supernatants removed from cells containing this gene
activated the EGR1 (early growth response gene 1) promoter element.
Thus, it is likely that this gene activates sensory neuron cells
and fibroblasts, in addition to other tissues or cell types,
through the EGR1 signal transduction pathway. The EGR1 (early
growth response gene 1) is a separate signal transduction pathway
from Jaks-STAT, genes containing the EGR1 promoter are induced in
various tissues and cell types upon activation, leading the cells
to undergo differentiation and proliferation.
[0131] Moreover, contact of cells with supernatant expressing the
product of this gene has been shown to increase the permeability of
the plasma membrane of monocytes to calcium. Thus it is likely that
the product of this gene is involved in a signal transduction
pathway that is initiated when the product-binds a receptor on the
surface of the plasma membrane of monocytes, in addition to other
cell-lines or tissue cell types, such as immune or hematopoietic
cells. Thus, polynucleotides and polypeptides have uses which
include, but are not limited to, activating monocytes.
[0132] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
ARAGQMQNLESARAGRSVSTQTGS (SEQ ID NO: 347). Polynucleotides encoding
these polypeptides are also encompassed by the invention. The gene
encoding the disclosed cDNA is believed to reside on chromosome 13.
Accordingly, polynucleotides related to this invention are useful
as a marker in linkage analysis for chromosome 13.
[0133] This gene is expressed primarily in T-cells.
[0134] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases involving immune regulation, which include,
but are not limited to autoimmune diseases such as rheumatoid
arthritis, lupus, and leukemia. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., hematopoietic,
immune, and cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0135] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 164 as residues: Ala-28 to His-41, Pro-43 to
Gln-64.
[0136] The tissue distribution in T-cells, combined with the
detected EGR1 and calcium flux activities, indicates
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders, particularly those dependent upon
signalling aberrations. Expression of this gene product in T-cells
indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of potentially all hematopoietic
cell lineages, including blood stem cells. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer--particularly considering the homology
to a conserved ras gene, and the detected EGR1 biological
activity.
[0137] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immune deficiency diseases such as
AIDS, and leukemia. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tumors and tissues. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0138] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:25 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1541 of SEQ ID NO:25, b is an integer
of 15 to 1555, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:25, and where b is greater
than or equal to a+14.
[0139] Features of Protein Encoded by Gene No: 16
[0140] This gene is expressed primarily in kidney cortex.
[0141] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the kidney including cancer and renal
dysfunction, in addition to, endocrine disorders, particularly of
the adrenal glands. Similarly, polypeptides and antibodies directed
to these polypeptides are useful in providing immunological probes
for differential identification of the tissue(s) or cell type(s).
For a number of disorders of the above tissues or cells,
particularly of the renal system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., urogenital, renal, endocrine,
and cancerous and wounded tissues) or bodily fluids (e.g., urine,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0142] The tissue distribution in kidney cortex indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment, diagnosis, and/or prevention of diseases
of the kidney including renal failure, nephritus, renal tubular
acidosis, proteinuria, pyuria, edema, pyelonephritis,
hydronephritis, nephrotic syndrome, crush syndrome,
glomerulonephritis, hematuria, renal colic and kidney stones, in
addition to Wilm's Tumor Disease, and congenital kidney
abnormalities such as horseshoe kidney, polycystic kidney, and
Falconi's syndrome. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0143] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:26 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1529 of SEQ ID NO:26, b is an integer
of 15 to 1543, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:26, and where b is greater
than or equal to a+14.
[0144] Features of Protein Encoded by Gene No: 17
[0145] This gene is expressed primarily in T-cell lymphoma, and to
a lesser extent, in bone marrow stromal cells.
[0146] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune or hematopoietic disorders, particularly
cancers, such as lymphomas and leukemias. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
immune, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0147] The tissue distribution in bone marrow stromal cells and
T-cell lymphoma indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the diagnosis, treatment,
and/or prevention of a variety of immune or hematopoietic
disorders. Expression of this gene product in T-cells indicates a
role in regulating the proliferation; survival; differentiation;
and/or activation of potentially all hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g., by boosting immune responses). Since the gene is
expressed in cells of lymphoid origin, the natural gene product may
be involved in immune functions. Therefore it may be also used as
an agent for immunological disorders including arthritis, asthma,
immune deficiency diseases such as AIDS, and leukemia. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tumors and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types.
[0148] Expression in bone marrow cells suggest that polynucleotides
and polypeptides corresponding to this gene are useful for the
treatment and diagnosis of hematopoetic related disorders such as
anemia, pancytopenia, leukopenia, thrombocytopenia or leukemia
since stromal cells are important in the production of cells of
hematopoietic lineages. The uses include bone marrow cell ex vivo
culture, bone marrow transplantation, bone marrow reconstitution,
radiotherapy or chemotherapy of neoplasia. The gene product may
also be involved in lymphopoiesis, therefore, it can be used in
immune disorders such as infection, inflammation, allergy,
immunodeficiency etc. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed tumors and
tissues.
[0149] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:27 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1248 of SEQ ID NO:27, b is an integer
of 15 to 1262, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:27, and where b is greater
than or equal to a+14.
[0150] Features of Protein Encoded by Gene No: 18
[0151] This gene is expressed primarily in medulloblastoma.
[0152] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, disorders of the central nervous system, including
cancers. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., neural, cancerous and wounded
tissues) or bodily fluids (e.g., serum, plasma, urine, synovial
fluid and spinal fluid) or another tissue or cell sample taken from
an individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0153] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 167 as residues: Phe-22 to Leu-28.
[0154] The tissue distribution indicates that polynucleotides and
polypeptides corresponding to this gene are useful for the
detection, treatment, and/or prevention of neurodegenerative
disease states and behavioural disorders such as Alzheimers
Disease, Parkinsons Disease, Huntingtons Disease, Tourette
Syndrome, schizophrenia, mania, dementia, paranoia, obsessive
compulsive disorder, panic disorder, learning disabilities, ALS,
psychoses , autism, and altered behaviors, including disorders in
feeding, sleep patterns, balance, and perception. In addition, the
gene or gene product may also play a role in the treatment and/or
detection of developmental disorders associated with the developing
embryo. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0155] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:28 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 739 of SEQ ID NO:28, b is an integer
of 15 to 753, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:28, and where b is greater
than or equal to a+14.
[0156] Features of Protein Encoded by Gene No: 19
[0157] The translation product of this gene was shown to have
homology to the mammalian notch I protein which has been shown to
be important in the regulation of cell-fate during pattern
formation and development (See Genbank Accession No.
gi.vertline.57635). One embodiment of this gene comprises
polypeptides of the following amino acid sequence:
8 KHEXHQVSDGALRCFASLADRFTRRGVDPAPLAKHGLTEE (SEQ ID NO: 348),
LLSRMAAAGGTVSGPSSACKPXRSTTGAPSTTADSKLSNQVSTIVSLLSTLCRG
SPVVTHDLLRSELPDSIESALQGDERCVLDTMRLVDFLLVLLFEGRKALPKSSA
GSTGRIPGLRRLDSSGERSHRQLIDCIRSKDTDALIDAIDTGAFEVNFMDDVG
QTLLNWASAFGTQEMVEFLCERGADVNRGQRSSSLHYAACFGRPQVAKT
LLRHGANPDLRDEDGKTPLDKARERGHSEVVAILQSPGDWMCPVNKGDDK
PLDKARERGHSEVVAIL (SEQ ID NO: 349), AKTLLRHGANPDLRD (SEQ ID NO:
350), GRGRAWLCRRPVGSWIGAVWNDKPDKETFKKPWQMWTQIHCWNG- YRWDXXDXKD (SEQ
ID NO: 351), SWIGAVWNDKPDKETFKKPWQMW (SEQ ID NO: 352),
KTMADVDPDTLLEWLQMGXGRXKGHATN TP (SEQ ID NO: 353),
RGVDPAPLAKHGLTEELLSRMAAAGGTVSGPSSA (SEQ ID NO: 354),
RSTTGAPSTTADSKLSNQVSTIVSLLSTLCR (SEQ ID NO: 355),
FEVNFMDDVGQTLLNWASAFGTQEMVEFLCERGA (SEQ ID NO: 356), and/or
EDGKTPLDKARERGHSEVVAILQSPGDW (SEQ ID NO: 357).
[0158] An additional embodiment is the polynucleotides encoding
these polypeptides.
[0159] This gene is expressed primarily in endothelial cells.
[0160] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, vascular disorders, particularly diseases involving
angiogenic abnormalities including diabetic retinopathy, macular
degeneration, and other diseases including arterioscerosis, stroke,
aneurysm, embolism, and cancer. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the vascular system, expression
of this gene at significantly higher or lower levels may be
routinely detected in certain tissues or cell types (e.g.,
endothelial, vascular, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue from
an individual not having the disorder.
[0161] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 168 as residues: Asp-17 to Phe-23.
[0162] The tissue distribution in endothelial cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for treating diseases where an increase or decrease in
angiogenesis is indicated and as a factor in the wound healing
process. The protein is useful in the treatment of cancer cells and
tissues, particularly in inhibiting angiogenesis of the invading
tumor. Protein, as well as, antibodies directed against the protein
may show utility as a tumor marker and/or immunotherapy targets for
the above listed tissues.
[0163] Alternatively, considering the homology to the Notch I
protein, this gene may show utility in the detection/treatment of
neurodegenerative disease states and behavioural disorders such as
Alzheimers Disease, Parkinsons Disease, Huntingtons Disease,
Tourette Syndrome, schizophrenia, mania, dementia, paranoia,
obsessive compulsive disorder, panic disorder, learning
disabilities, ALS, psychoses, autism, and altered behaviors,
including disorders in feeding, sleep patterns, balance, and
perception. In addition, the gene or gene product may also play a
role in the treatment and/or detection of developmental disorders
associated with the developing embryo, sexually-linked disorders,
or disorders of the cardiovascular system. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0164] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:29 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1607 of SEQ ID NO:29, b is an integer
of 15 to 1621, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:29, and where b is greater
than or equal to a+14.
[0165] Features of Protein Encoded by Gene No: 20
[0166] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: TRPTMPNFLWFPKCA (SEQ ID
NO: 358). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0167] This gene is expressed primarily in meningioma tissues.
[0168] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neural or central nervous system disorders,
particularly cancers of the central nervous system and endothelium.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., neural, endothelial, CNS, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0169] The tissue distribution in meningioma tissue indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses,
autism, and altered behaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing embryo,
sexually-linked disorders, or disorders of the cardiovascular
system.
[0170] Moreover, the protein is useful in inhibiting or
ameliorating infections of the meninges, particular viral
infections. In addition, the protein may show utility in the
treatment, detection, and/or prevention of such infections and
disorders, in addition to degenerative conditions or congenital
defects of the meninges. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0171] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:30 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 907 of SEQ ID NO:30, b is an integer
of 15 to 921, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:30, and where b is greater
than or equal to a+14.
[0172] Features of Protein Encoded by Gene No: 21
[0173] The translation product of this gene was shown to have
homology to the retinoic acid receptor gamma-2 which is thought to
be important in the development of, and may be a key determinant
for, human breast cancer during aberrant activation (See Genbank
Accession No. AA176435). In specific embodiments, polypeptides of
the invention comprise the following amino acid sequence:
9 LPPCLAQIFPFFSSGTNLTFCFFVFVFVFVFAELDYRNSYEIEY (SEQ ID NO:
359).
[0174] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0175] This gene is expressed primarily in ovary, and to a lesser
extent, in meningioma.
[0176] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive or neural disorders, particularly ovarian
cancer, as well as, other cancers of the reproductive system,
meninges, and endothelial tissue in general. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the female
reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., ovarian, reproductive, neural, endothelial,
endocrine, and cancerous and wounded tissues) or bodily fluids
(e.g., amniotic fluid, lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0177] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 170 as residues: Leu-8 to Gln-1 8, Thr-26 to Lys-33,
Met-39 to Cys-46, Ala-62 to Pro-69, Pro-83 to Glu-90.
[0178] The tissue distribution in ovarian tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for diagnosis and intervention of tumors within these
tissues, in addition to other tumors where expression has been
indicated. The protein may also show utility in the treatment,
detection, prevention, and/or amelioration of degenerative
conditions or congenital disorders of the meninges, and the brain
and spinal cord, in general. Protein, as well as, antibodies
directed against the protein may show utility as a tissue-specific
marker and/or immunotherapy target for the above listed tumors and
tissues.
[0179] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:31 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 2081 of SEQ ID NO:31, b is an integer
of 15 to 2095, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:3 1, and where b is greater
than or equal to a+14.
[0180] Features of Protein Encoded by Gene No: 22
[0181] This gene is expressed primarily in the spongy tissue from
Alzheimer's brain.
[0182] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neural disorders, which include, but are not limited to
Alzheimer's disease and other neurodegenerative diseases.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells. particularly of
the central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., neural, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0183] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 171 as residues: Ser-31 to Ala-37, Ala-50 to Tyr-55,
Phe-63 to Arg-68, His-83 to Pro-89.
[0184] The tissue distribution in spongy tissue from Alzheimer's
patient indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the detection/treatment
of neurodegenerative disease states and behavioural disorders such
as Alzheimers Disease, Parkinsons Disease, Huntingtons Disease,
Tourette Syndrome, schizophrenia, mania, dementia, paranoia,
obsessive compulsive disorder, panic disorder, learning
disabilities, ALS, psychoses , autism, and altered behaviors,
including disorders in feeding, sleep patterns, balance, and
perception. In addition, the gene or gene product may also play a
role in the treatment and/or detection of developmental disorders
associated with the developing embryo. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0185] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:32 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1824 of SEQ ID NO:32, b is an integer
of 15 to 1838, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:32, and where b is greater
than or equal to a+14.
[0186] Features of Protein Encoded by Gene No: 23
[0187] This gene is expressed primarily in bone marrow cells.
[0188] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune or hematological disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
hematological and immune systems, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, haematopoeitic, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0189] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 172 as residues: Glu-22 to Ser-33, Leu-47 to Ser-55,
Thr-87 to Arg-104.
[0190] The tissue distribution in bone marrow cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment and diagnosis of hematopoetic related
disorders, which include, but are not limited to anemia,
pancytopenia, leukopenia, thrombocytopenia or leukemia since
stromal cells are important in the production of cells of
hematopoietic lineages. The uses include bone marrow cell ex vivo
culture, bone marrow transplantation, bone marrow reconstitution,
radiotherapy or chemotherapy of neoplasia. The gene product may
also be involved in lymphopoiesis, therefore, it can be used in
immune disorders such as infection, inflammation, allergy,
immunodeficiency etc. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0191] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:33 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 768 of SEQ ID NO:33, b is an integer
of 15 to 782, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:33, and where b is greater
than or equal to a+14.
[0192] Features of Protein Encoded by Gene No: 24
[0193] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: LKCTIYGGA (SEQ ID NO:
360). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0194] This gene is expressed primarily in neutrophils.
[0195] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the immune system, including inflammatory
diseases and allergies. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune,
haematopoeitic, and. cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0196] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 173 as residues: Gln-36 to Lys-41.
[0197] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product
indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g. by boosting immune responses). Since the gene is
expressed in cells of lymphoid origin, the natural gene product may
be involved in immune functions.
[0198] Therefore it may be also used as an agent for immunological
disorders including arthritis, asthma, immunodeficiency diseases
such as AIDS, leukemia, rheumatoid arthritis, granulomatous
disease, inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues, such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic, anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0199] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:34 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1546 of SEQ ID NO:34, b is an integer
of 15 to 1560, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:34, and where b is greater
than or equal to a+14.
[0200] Features of Protein Encoded by Gene No: 25
[0201] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
10 HVLWSLLSACWTQFLVYFCCLMILQRTFPPRALRTSPWLSNPMGVKGKKKKGTFME (SEQ ID
NO: 361), and/or ELVYFCCLMILQRTFPPRALRTSPWLSNPM (SEQ ID NO:
362).
[0202] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0203] This gene is expressed primarily in neutrophils.
[0204] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune or hematopoietic disorders, including
inflammatory diseases and allergies. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune,
haematopoeitic, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0205] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product
indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g. by boosting immune responses).
[0206] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immunodeficiency diseases such as
AIDS, leukemia, rheumatoid arthritis, granulomatous disease,
inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues, such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0207] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:35 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence-would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1078 of SEQ ID NO:35, b is an integer
of 15 to 1092, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:35, and where b is greater
than or equal to a+14.
[0208] Features of Protein Encoded by Gene No: 26
[0209] This gene is expressed primarily in neutrophils.
[0210] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune or hematopoietic disorders, including
inflammatory conditions and allergies. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune,
hematopoietic, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0211] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 175 as residues: Lys-9 to Leu-16, Ser-33 to
Met-43.
[0212] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product
indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g. by boosting immune responses).
[0213] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immunodeficiency diseases such as
AIDS, leukemia, rheumatoid arthritis, granulomatous disease,
inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0214] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:36 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1139 of SEQ ID NO:36, b is an integer
of 15 to 1153, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:36, and where b is greater
than or equal to a+14.
[0215] Features of Protein Encoded by Gene No: 27
[0216] The translation product of this gene was shown to have
homology to the intrinsic factor-B 12 receptor precursor of Rattus
norvegicus which is thought to be important in development (See
Genbank Accession No. gi.vertline.2961490 (AF022247)). One
embodiment of this gene comprises polypeptides of the following
amino acid sequence:
11 DCNRDYHKAFGNLRSPGWPDNYDNDXDCXVTLTAPQNHHSGIVENAETISWR (SEQ ID NO:
363), FGNLRSPGWPDNYDN (SEQ ID NO: 364), ASFYRTS (SEQ ID NO: 366),
and/or APQNHXLKCRNDFLEV (SEQ ID NO: 365).
[0217] An additional embodiment is the polynucleotides encoding
these polypeptides.
[0218] This gene is expressed primarily in neutrophils.
[0219] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune or hematopoietic disorders, including
inflammatory disorders, and allergies. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune,
haematopoetic, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0220] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product
indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g. by boosting immune responses).
[0221] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immunodeficiency diseases such as
AIDS, leukemia, rheumatoid arthritis, granulomatous disease,
inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues, such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0222] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:37 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 971 of SEQ ID NO:37, b is an integer
of 15 to 985, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:37, and where b is greater
than or equal to a+14.
[0223] Features of Protein Encoded by Gene No: 28
[0224] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
12 KADVKWHMCLQSPLCGLFCSIEGVLK (SEQ ID NO: 367),
ACMNPAMCFVCACPHTGSTPEKAILQGRLISLGTSLSPASNGSGQQSFSICMI (SEQ ID NO:
368), NPSLPXSTSSHHLFSVLTGDLDSYSQRKLKPTSRKSFLLPKTQTYXVXHPSSP
PLVLVQHRSPLSTYPKPVPSCCALDLISVIALETFLVYIHLFPSIDLSYWILSMLQ
PLLLIKQQSTKTLSLNCMLYSSYYLISFLSFKAKVLRRGGNILHHFFTSYSEFNTY
CPHTGSTPEKAILQGRLISLGTSLSPAS (SEQ ID NO: 369),
QHRSPLSTYPKPVPSCCALDLISV (SEQ ID NO: 370), and/or
IKQQSTKTLSLNCMLYSSYYLISFLSFKA (SEQ ID NO: 371).
[0225] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0226] This gene is expressed primarily in neutrophils.
[0227] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune and/or haematological disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., immune, hematopoietic and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0228] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 177 as residues: Pro-55 to Ser-66.
[0229] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product
indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g. by boosting immune responses).
[0230] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immunodeficiency diseases such as
AIDS, leukemia, rheumatoid arthritis, granulomatous disease,
inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues, such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0231] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:38 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1108 of SEQ ID NO:38, b is an integer
of 15 to 1122, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:38, and where b is greater
than or equal to a+14.
[0232] Features of Protein Encoded by Gene No: 29
[0233] This gene is expressed primarily in neutrophils.
[0234] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune and haematological disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., immune, hematopoietic, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0235] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product
indicates a role in regulating the proliferation; survival;
differentiation; and/or activation of hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g. by boosting immune responses).
[0236] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immunodeficiency diseases such as
AIDS, leukemia, rheumatoid arthritis, granulomatous disease,
inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues, such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0237] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:39 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 584 of SEQ ID NO:39, b is an integer
of 15 to 598, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:39, and where b is greater
than or equal to a+14.
[0238] Features of Protein Encoded by Gene No: 30
[0239] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
KYLVSSVLPTISMARSLISALRSG (SEQ ID NO: 372). Polynucleotides encoding
these polypeptides are also encompassed by the invention. The gene
encoding the disclosed cDNA is believed to reside on chromosome 7.
Accordingly, polynucleotides related to this invention are useful
as a marker in linkage analysis for chromosome 7.
[0240] This gene is expressed primarily in ovarian cancer.
[0241] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive disorders, particularly ovarian cancer.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., reproductive, ovarian, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0242] The tissue distribution in ovarian tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for diagnosis and treatment of ovarian cancer. Moreover, the
protein is useful for the treatment, detection, and/or prevention
of endocrine disorders, particularly those related to the
reproductive system. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0243] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:40 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1115 of SEQ ID NO:40, b is an integer
of 15 to 1129, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:40, and where b is greater
than or equal to a+14.
[0244] Features of Protein Encoded by Gene No: 31
[0245] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
13 MRTLFGAVRAPFSSLTLLLITPSPSPL (SEQ ID NO: 373), MAYAFHRTST (SEQ ID
NO: 374), LKSTYTLLSILWFLVLIPVEGN (SEQ ID NO: 375), and/or
GPLLASHATLCFSLGSKF (SEQ ID NO: 376).
[0246] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0247] This gene is expressed primarily in ovarian cancer.
[0248] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive, or endocrine disorders, particularly
proliferative condtions such as ovarian cancer. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., reproductive, endocrine, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0249] The tissue distribution in ovarian tumor tissue indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for diagnosis and treatment of ovarian cancer. Moreover,
the protein is useful for the detection, treatment, and/or
prevention of a variety of reproductive disorders such as
infertility. In addition, the protein may also be useful in the
development of novel or improved contraceptives. The expression
within cellular sources marked by proliferating cells indicates
this protein may play a role in the regulation of cellular
division, and may show utility in the diagnosis and treatment of
cancer and other proliferative disorders. Similarly, developmental
tissues rely on decisions involving cell differentiation and/or
apoptosis in pattern formation. Thus this protein may also be
involved in apoptosis or tissue differentiation and could again be
useful in cancer therapy. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0250] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:41 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1144 of SEQ ID NO:41, b is an integer
of 15 to 1158, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:41, and where b is greater
than or equal to a+14.
[0251] Features of Protein Encoded by Gene No: 32
[0252] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: TVWGILPRKR (SEQ ID NO:
377). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0253] This gene is expressed primarily in ovarian tumor.
[0254] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive or endocrine disorders, particularly
proliferative conditions such as ovarian cancer. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., reproductive, endocrine, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0255] The tissue distribution in ovarian tumor tissue indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for diagnosis and treatment of ovarian cancer. Moreover,
the protein is useful for the detection, treatment, and/or
prevention of a variety of reproductive disorders such as
infertility. In addition, the protein may also be useful in the
development of novel or improved contraceptives. The expression
within cellular sources marked by proliferating cells indicates
this protein may play a role in the regulation of cellular
division, and may show utility in the diagnosis and treatment of
cancer and other proliferative disorders. Similarly, developmental
tissues rely on decisions involving cell differentiation and/or
apoptosis in pattern formation. Thus this protein may also be
involved in apoptosis or tissue differentiation and could again be
useful in cancer therapy. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0256] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:42 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1753 of SEQ ID NO:42, b is an integer
of 15 to 1767, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:42, and where b is greater
than or equal to a+14.
[0257] Features of Protein Encoded by Gene No: 33
[0258] The translation product of this gene shares sequence
homology with uroplakin III which is thought to be important in
urothelial differentiation (See Accession No. d10226610). In
specific embodiments, polypeptides of the invention comprise the
following amino acid sequence: ASIDTWPGRRSGGMIVITSI (SEQ ID NO:
378) and/or GSPQAETRWSDPIALHQGKSPASIDTWP- GRRSGGMIVITSI (SEQ ID NO:
379). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0259] This gene is expressed primarily in ovarian tumor.
[0260] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive or endocrine disorders, particularly
proliferative conditions such as ovarian cancer. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., reproductive, endocrine, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0261] The tissue distribution in ovarian tumor tissue, combined
with the homology to uroplakin III indicates that polynucleotides
and polypeptides corresponding to this gene are useful for the
diagnosis and treatment of reproductive disorders, urogential
condtions, or endocrine disorders. Moreover, the protein is useful
for the detection, treatment, and/or prevention of a variety of
reproductive disorders such as infertility. In addition, the
protein may also be useful in the development of novel or improved
contraceptives. The expression within cellular sources marked by
proliferating cells indicates this protein may play a role in the
regulation of cellular division, and may show utility in the
diagnosis and treatment of cancer and other proliferative
disorders. Similarly, developmental tissues rely on decisions
involving cell differentiation and/or apoptosis in pattern
formation. Thus this protein may also be involved in apoptosis or
tissue differentiation and could again be useful in cancer therapy.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0262] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:43 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 903 of SEQ ID NO:43, b is an integer
of 15 to 917, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:43, and where b is greater
than or equal to a+14.
[0263] Features of Protein Encoded by Gene No: 34
[0264] The translation product of this gene shares sequence
homology with estrogen-responsive finger protein. which is thought
to be important in uterine implantation. (See Accession No.
1088467; and J. Biol. Chem. 270 (41), 24406-24413 (1995), herein
incorporated by reference in its entirety.) Moreover, the protein
product of this gene was also shown to homology to the human rfp
transforming protein (See Genbank Accession No. gil337372) which is
thought to play a role in in male germ cell development. Preferred
polypeptide fragments comprise the amino acid sequence:
14 VXDITFDPDTAHKYLRLQEENRKVTNTTPWEHPYPDLPSRFLH (SEQ ID NO: 380);
LYLHRYYFEVEIFGAGTYV (SEQ ID NO: 381); SCISGNNFSWSLQWNGKEFTAW (SEQ
ID NO: 382); TPLKAGPFWSSGSILTS (SEQ ID NO: 383);
SVSEVKAVAEMQFGELLAAVRKAQANVMLFLXEKEQAAL (SEQ ID NO: 384);
EKSKQELETMAAISNTVQFLEEYCKFKNTEDITFPSVYIGLKD (SEQ ID NO: 385);
LENYKKKLQEFSKEEEYDIRTQVSAXVQR (SEQ ID NO: 386); GTVSRERRAG (SEQ ID
NO: 388), HGDPTQSWPFLELGVYIDFPGGILSFYG- VEYDSM (SEQ ID NO: 389),
TLVHKFACKFSEPVYAAFWLSKKENAIRIVDLG- EEPEKPAP SLVGTAP
SFYGVEYDSMTLVHKFACKFSEPVYAAFWL (SEQ ID NO: 390),
AELQCTQLDLERKLKLNENAISRLQANQKSVLVSVSEVKAVAEMQFG- ELLAAV (SEQ ID NO:
391), RKAQANVMLFLXEKEQAALSQANGIKAHLEYKS- AEMEKSKQELETMAAISN
TVQFLEEYCKFKNTEDITFPSVYIGLKDKLSGIRKVITE- STVHLIXXLENYKKKL
QEFSKEEEYDIRTQVSAXVQRKYWTSKPEPSTREQFLQYVX- DITFDPDTAHKYL
RLQEENRKVTNTTPWEHPYPDLPSRFLHWRQVLSQQSLYLHRYY- FEVEIFGA
GTYVGLTCKGIDXKGEERXSCISGNNFSWSLQWNGKEFTAWYSDMETPL
KAGPFWSSGSILTSQEGSFPSMA RTAPYGAKESSWR (SEQ ID NO: 392), and/or
MFSFRDPIGFQKPATISSYFCPQITLKCKSHH- CSWQRSGIWLLESREQSPPRT
VLASRVPLPDLQSGWRFPSWKARRQHRLVLKTCRQT- CEPESWNHTLRHRR
KGSLLGSQYRPRAPERASFEWGLHVTVPGRELLPVPLEAPGEV- VSGNATXALL
PFXVDAFAGQANIGACPEDLHLKIVPVQVQTLLGQHLPPVQEPAGEV- RVG
MLPGRGVGDLAVLLLQPEILVCCVRVERDVXHILEELFPGAGLRFGSPIFALN
NGRHLSSDVILLFLGKLLELFLIVLQXXD
GVYIDFPGGILSFYGVEYDSMTLVHKFACKFSEPVYAA (SEQ ID NO: 387).
[0265] Also preferred are polynucleotide fragments encoding these
polypeptide fragments.
[0266] This gene is expressed primarily in ovarian cancer.
[0267] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, ovarian cancer and other disorders of the reproductive
system. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., reproductive, developmental, ovarian,
testicular, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, seminal, fluid, amniotic fluid, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0268] The tissue distribution in ovarian tumors, combined with the
homology to estrogen-responsive finger protein, in addition, to the
conserved rfp transforming protein indicates that polynucleotides
and polypeptides corresponding to this gene are useful for
diagnosis and treatment of ovarian cancer and other disorders of
the reproductive system. Moreover, the expression within cellular
sources marked by proliferating cells indicates this protein may
play a role in the regulation of cellular division, and may show
utility in the diagnosis and treatment of cancer and other
proliferative disorders. Similarly, developmental tissues rely on
decisions involving cell differentiation and/or apoptosis in
pattern formation. Thus this protein may also be involved in
apoptosis or tissue differentiation and could again be useful in
cancer therapy. The protein may also show utility in the
development of novel contraceptives. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0269] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:44 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1973 of SEQ ID NO:44, b is an integer
of 15 to 1987, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:44, and where b is greater
than or equal to a+14.
[0270] Features of Protein Encoded by Gene No: 35
[0271] This gene shows sequence homology to a Caenorhabditis
elegans gene, called D1054.3, in addition, to the Sgt1p protein of
Saccharomyces cerevisiae which are thought to play a role in the
regulation of cellular division and developmental precesses (See
Accession Nos. gn1.vertline.PID.vertline.e348554 and
gi.vertline.1870791, respectively) Preferred polypeptide fragments
comprise the amino acid sequence:
15 SKIKYDWYQTESQVVITLMIKNVQKNDVNVEFSEKELSALVKLPSGEDYNLKL (SEQ ID
NO: 393); ELLHPIIPEQSTFKVLSTKIEIKLKKPEAVRWEKLEGQGDVPTPKQ- FVDVKNLY
PSSSPTRNWDKLVGEIKEEEKNEKLEGDAALNRLFQQIYSDGSDEVKRA- MN
KSFMESGGTVLSTNWSDVGKRKVEINPPDDMEWKKY
GDAALNRLFQQIYSDGSDEVKRAMNKSFMESGGTVLSTN (SEQ ID NO: 394);
MAAAAAGTXXSQRFFQSFSDALIDEDPQAALEELTKALEQKPDDAQYYCQ (SEQ ID NO:
396), RAYCHILLGNYCVAVADAKKSLELNPNNSTAMLRKGICEYHEKNYAAALETFT
EGQKLDSADAN FSVWIKRCQEAQNGSESEVVSPKFSFFMFLLF LEELTKALEQKPDD
AQYYCQRAYCHILLGNYCVAVADA (SEQ ID NO: 397),
AMLRKGICEYHEKNYAAALETFTEGQKLDSA (SEQ ID NO: 398), LRLWNRNQMM
HSIIVKELIVTFFLGITVLLLLMQRSL (SEQ ID NO: 399), NSIQIIPLLC (SEQ ID
NO: 400), YMHFNNTVAKLTCKNLSLSTYQNQSASQ- WTHQSKIKYDW (SEQ ID NO:
401), YQTESQVVITLMIKNVQKNDVNVEFSEK- ELSALVKLPSGEDYNLKLELLHPI
IPEQSTFKVLSTKIEIKLKKPEAVRWEKLEGQG- DVPTPKQFVADVKNLYPSSS
PYTRNWDKLVGEIKEEEKNEKLEGDAALNRLFQQIYS- DGSDEVKRAMNKSF
MESGGTVLSTNWSDVGKRKVEINPPDDMEWKKY TCKNLSLSTYQNQSASQWTHQSKIKYDWY
(SEQ ID NO: 402), EKELSALVKLPSGEDYNLKLELLH (SEQ ID NO: 403),
LHPIIPEQSTFKVLSTKIEIKLKKPEAVR (SEQ ID NO: 404),
KQFVADVKNLYPSSSPYTRNWDKL (SEQ ID NO: 405), and/or
DWYQTESQVVITLMIKNVQKNDV (SEQ ID NO: 395).
[0272] Also preferred are polynucleotide fragments encoding these
polypeptide fragments.
[0273] This gene is expressed primarily in osteoclastoma.
[0274] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include; but are not
limited to, skeletal or developmental disorders, particularly
osteoclastoma and other forms of Cancer. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the skeletal system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
skeletal, developmental, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0275] The tissue distribution in osteoclastoma, combined with the
homology to the D1054.3 and Sgt1p proteins indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for diagnosis and treatment of osteoclastoma and other forms
of cancers. Moreover, the expression within embryonic tissue and
other cellular sources marked by proliferating cells indicates this
protein may play a role in the regulation of cellular division, and
may show utility in the diagnosis and treatment of cancer and other
proliferative disorders. Similarly, developmental tissues rely on
decisions involving cell differentiation and/or apoptosis in
pattern formation. Thus this protein may also be involved in
apoptosis or tissue differentiation and could again be useful in
cancer therapy. Protein may also play a role as a tumor supressor,
or in the development of tumor progression inhibitors. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0276] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:45 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 2039 of SEQ ID NO:45, b is an integer
of 15 to 2053, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:45, and where b is greater
than or equal to a+14.
[0277] Features of Protein Encoded by Gene No: 36
[0278] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
16 GSKGQERKWRVRMGYLN (SEQ ID NO: 406),
QRYRLLPLFCYVCSRKIKLNENLFVFSAYSLATLPHTYLFSIVEC SSFCLSGTRN (SEQ ID
NO: 407), and/or FSAYSLATLPHTYLFSIVEC SSFCLSG (SEQ ID NO: 408).
[0279] Polynucleotides encoding these polypeptides are also
encompassed by the invention. The gene encoding the disclosed cDNA
is believed to reside on chromosome 7. Accordingly, polynucleotides
related to this invention are useful as a marker in linkage
analysis for chromosome 7.
[0280] This gene is expressed primarily in human placenta.
[0281] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, developmental, vascular, and/or reproductive disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the embryonic and reproductive systems, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., developmental, vascular,
reproductive, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, amniotic fluid, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0282] The tissue distribution in human placenta tissue indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for the treatment and diagnosis of the disorders of
embryonic and reproductive systems. Moreover, the protein is useful
for the detection, treatment, and/or prevention of a variety of
vascular disorders, which include, but are not limited to,
microvascular disease, aneurysm, arteriosclerosis, atherosclerosis,
stroke, or embolism. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0283] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:46 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1258 of SEQ ID NO:46, b is an integer
of 15 to 1272, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:46, and where b is greater
than or equal to a+14.
[0284] Features of Protein Encoded by Gene No: 37
[0285] This gene is expressed primarily in anergic T-cells.
[0286] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune or hematopoietic disorders, particularly
inflammatory conditions and immunodeficiencies such as AIDS.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune system, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types (e.g., immune, hematopoietic, and cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0287] The tissue distribution in anergic T-cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment and diagnosis of T cell related disorders.
Moreover, the expression of this gene product indicates a role in
regulating the proliferation; survival; differentiation; and/or
activation of hematopoietic cell lineages, including blood stem
cells. This gene product may be involved in the regulation of
cytokine production, antigen presentation, or other processes that
may also suggest a usefulness in the treatment of cancer (e.g. by
boosting immune responses).
[0288] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immunodeficiency diseases such as
AIDS, leukemia, rheumatoid arthritis, granulomatous disease,
inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues, such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0289] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:47 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 759 of SEQ ID NO:47, b is an integer
of 15 to 773, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:47, and where b is greater
than or equal to a+14.
[0290] Features of Protein Encoded by Gene No: 38
[0291] The translation product of this gene shares sequence
homology with a murine bone-related sulphatase (See Genbank
Accession No. 3046314, and Genseq Accession No. R51355) which is
thought to be involved in proteoglycan metabolism. In specific
embodiments, polypeptides of the invention comprise the following
amino acid sequence:
17 ASFGSCSLSLPCSARERTPEGGGWPGGRLSEPLPA (SEQ ID NO: 409), APNVVLV
(SEQ ID NO: 410), DGRLTF (SEQ ID NO: 411), PGSQVVKLPFINFM (SEQ ID
NO: 412), FLNAYTNSP (SEQ ID NO: 413), ICCPSRAAMWSGLFTHLTESWNNFKG-
LDPNYTTWMD (SEQ ID NO: 414), TQKFGK (SEQ ID NO: 415), DYTSGHHSI
(SEQ ID NO: 416), SNRVEAWTRDVAFLLRQEGRP (SEQ ID NO: 417), DWQNTDKA
(SEQ ID NO: 418), YLGLNLPHPYPSPSSGENFGSSTFHTSLYWLEKV (SEQ ID NO:
419), DAIKIPKW (SEQ ID NO: 420), YTKNCTG (SEQ ID NO: 421),
NIRAFYYAMCAETDAMLGEIILALH (SEQ ID NO: 422), LDLLQKTIVIY (SEQ ID NO:
423), MEHRQFYKMSMYEAS (SEQ ID NO: 424), HVPLLMMGPGIKA (SEQ ID NO:
425), VVSLVDIYPTMLDIAGI (SEQ ID NO: 426), DPDELTN (SEQ ID NO: 427),
WKYIAY (SEQ ID NO: 428), NFPEITYSLDQKLHSIINYPKVSASVHQY-
NKEQFIKWKQSIGQNYSNVIANFRWHQDWOKEPRKYENAID (SEQ ID NO: 429),
QWLKTHMNPRAV FPEITYSLDQKL (SEQ ID NO: 430), NYPKVSASVHQYNKEQFI (SEQ
ID NO: 431), GQNYSNVIA (SEQ ID NO: 432), RWHQDWQ (SEQ ID NO: 433),
and/or PRKYENAI (SEQ ID NO: 434).
[0292] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0293] This gene is expressed primarily in retina.
[0294] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, visual, skeletal, or metabolic disorders, particularly
eye dieases and bone metabolic disorders. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the eye, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., visual, skeletal,
metabolic, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, vitreous humor, aqueous humor,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0295] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 187 as residues: Ala-21 to Arg-27, Asp-40 to Arg-45,
Glu-97 to Thr-110, Glu-117 to Lys-128, Arg-175 to Lys-182, Pro-207
to Gly-220, Val-253 to Ile-272.
[0296] The tissue distribution in retina, combined with the
homology to sulphatases indicates that polynucleotides and
polypeptides corresponding to this gene are useful for diagnosis
and treatment of eye disorders. Moreover, this gene may be useful
in the detection, treatment, and/or prevention of bone-related
disorders, osteoporosis, Paget's disease, osteomalacia, in addition
to bone metabolic disorders, particularly those involving
proteoglycans. The protein is also useful in the disorders
involving aberrant proteoglycan metabolism or related conditions,
which may include arthritis, immune cell migration, cellular
proliferation, vascular disorders, hematopoietic disorders, in
addition to showing utility in the detection, treatment, and/or
prevention of the disorders mentioned above. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0297] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:42 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 2105 of SEQ ID NO:48, b is an integer
of 15 to 2119, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:48, and where b is greater
than or equal to a+14.
[0298] Features of Protein Encoded by Gene No: 39
[0299] This gene is expressed primarily in human stomach
cancers.
[0300] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, gastointestinal disorders, particularly cancer.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the cancer, expression of this gene at significantly higher or
lower levels may be routinely detected in certain tissues or cell
types (e.g., endothelial, gastrointestinal, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, chyme, synovial fluid and spinal fluid) or another tissue or
cell sample taken from an individual having such a disorder,
relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0301] The tissue distribution in tumors of the stomach indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for the diagnosis, treatment, and/or prevention of these
tumors, in addition to other tumors in other tissues. The protein
may also be useful for the treatment and/or prevention of ulcers,
in addition to additional gastrointestinal or metabolic conditions.
Protein, as well as, antibodies directed against the protein may
show utility as a tissue-specific marker and/or immunotherapy
target for the above listed tissues.
[0302] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:49 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1174 of SEQ ID NO:49, b is an integer
of 15 to 1188, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:49, and where b is greater
than or equal to a+14.
[0303] Features of Protein Encoded by Gene No: 40
[0304] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: RNSLHCYNEQPPNASGLIQWSSD
LIPISLQCGCSW (SEQ ID NO: 435). Polynucleotides encoding these
polypeptides are also encompassed by the invention.
[0305] This gene is expressed primarily in human synovial
membrane.
[0306] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of synovial membrane, skeletal and/or
musculoskeletal disorders. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the synovial membrane system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
skeletal, muscular, rheumatiod, synovial, and cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0307] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 189 as residues: Pro-10 to Ser-20.
[0308] The tissue distribution in synovial tissue indicates the
product of this gene may play a role in the detection, treatment,
and/or prevention of disorders and conditions affecting the
skeletal systemskeletal system, in particular osteoporosis, bone
cancer, as well as, disorders afflicting connective tissues (e.g.
arthritis, trauma, tendonitis, chrondomalacia and inflammation),
such as in the diagnosis or treatment of various autoimmune
disorders such as rheumatoid arthritis, lupus, scleroderma, and
dermatomyositis as well as dwarfism, spinal deformation, and
specific joint abnormalities as well as chondrodysplasias (i.e.
spondyloepiphyseal dysplasia congenita, familial osteoarthritis,
Atelosteogenesis type II, metaphyseal chondrodysplasia type
Schmid). Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0309] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:50 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 464 of SEQ ID NO:50, b is an integer
of 15 to 478, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:50, and where b is greater
than or equal to a+14.
[0310] Features of Protein Encoded by Gene No: 41
[0311] The translation product of this gene shares sequence
homology with adipose specific collagen-like factor as well as the
human adipocyte complement related protein Acrp30, the latter of
which is known to be important in energy balance and homeostasis
involving food intake, particularly in carbohydrate and lipid
catabolism/anabolism (See Genbank Accession
Nos.gnl.vertline.PID.vertline.d1008822 and W09108, respectively).
One embodiment of this gene comprises polypeptides of the following
amino acid sequence:
18 XLWDPGLPGVCRCGSIVLKSAFSVGITTSYPEXRLPIIFNKVLLPRGXALQPC (SEQ ID
NO: 436), HRGSSSVLSQGIYYFSYDITLANKHLAIGLVHNGQYRIKTFDANTG- NHDVASG
STVIYLQPEDEVWLEIFFTDQNGLFSDPGWADSLFSGFLLYVDTDYLDSI- SEDDEL
GSIVLKSAFSVGITT (SEQ ID NO: 437), GIYYFSYDITLANK (SEQ ID NO: 438),
DSLFSGFLLYVDT (SEQ ID NO: 439), and/or NHDVASGSTVIYL (SEQ ID NO:
440).
[0312] An additional embodiment is the polynucleotides encoding
these polypeptides.
[0313] This gene is expressed primarily in human schwanoma.
[0314] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neural or integumentary disorders, particularly
neurofibroma. Similarly, polypeptides and antibodies directed to
these polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the diseases relating to peripheral or sympathetic nervous system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
neural, integumentary, extracellular matrix, cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0315] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 190 as residues: Gly-16 to Pro-30, Pro-42 to Gly-56,
Gly-62 to Gly-77, Glu-93 to Gly-104, Glu-109 to Glu-114, Pro-121 to
Asp-126.
[0316] The tissue distribution in schwanoma cells combined with the
homology to a conserved human adipose specific collagen-like factor
as well as to the human adipocyte complement related protein
Acrp30, indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the detection/treatment
of neurodegenerative disease states and behavioural disorders
particularly neuroschwannoma, and including Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses ,
autism, and altered behaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing embryo,
sexually-linked disorders, or disorders of the cardiovascular
system.
[0317] Moreover, polynucleotides and polypeptides corresponding to
this gene are useful for the treatment, diagnosis, and/or
prevention of various skin disorders including congenital disorders
(i.e. nevi, moles, freckles, Mongolian spots, hemangiomas,
port-wine syndrome), integumentary tumors (i.e. keratoses, Bowen's
disease, basal cell carcinoma, squamous cell carcinoma, malignant
melanoma, Paget's disease, mycosis fungoides, and Kaposi's
sarcoma), injuries and inflammation of the skin (i.e. wounds,
rashes, prickly heat disorder, psoriasis, dermatitis),
atherosclerosis, uticaria, eczema, photosensitivity, autoimmune
disorders (i.e. lupus erythematosus, vitiligo, dermatomyositis,
morphea, scleroderma, pemphigoid, and pemphigus), keloids, striae,
erythema, petechiae, purpura, and xanthelasma. In addition, such
disorders may predispose increased susceptibility to viral and
bacterial infections of the skin (i.e. cold sores, warts,
chickenpox, molluscum contagiosum, herpes zoster, boils,
cellulitis, erysipelas, impetigo, tinea, althletes foot, and
ringworm). Moreover, the protein product of this gene may also be
useful for the treatment or diagnosis of various connective tissue
disorders such as arthritis, trauma, tendonitis, chrondomalacia and
inflammation, autoimmune disorders such as rheumatoid arthritis,
lupus, scleroderma, and dermatomyositis as well as dwarfism, spinal
deformation, and specific joint abnormalities as well as
chondrodysplasias (i.e. spondyloepiphyseal dysplasia congenita,
familial osteoarthritis, Atelosteogenesis type II, metaphyseal
chondrodysplasia type Schmid).
[0318] Alternatively, considering the homology to a conserved
adipose specific collagen-like factor, would suggest that this
protein may also be important in the diagnosis or treatment of
various autoimmune disorders such as rheumatoid arthritis, lupus,
scleroderma, and dermatomyositis as well as dwarfism, spinal
deformation, and specific joint abnormalities as well as
chondrodysplasias ie. spondyloepiphyseal dysplasia congenita,
familial osteoarthritis, Atelosteogenesis type II, metaphyseal
chondrodysplasia type Schmid. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0319] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:51 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1319 of SEQ ID NO:51, b is an integer
of 15 to 1333, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:51, and where b is greater
than or equal to a+14.
[0320] Features of Protein Encoded by Gene No: 42
[0321] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: SNSHTHTHVKSFLR (SEQ ID
NO: 441). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0322] This gene is expressed primarily in human activated
T-Cells.
[0323] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immunodeficiencies, inflammatory conditions, and other
immune or hematopoietic disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the disorders of the immune
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., immune, hematopoietic, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0324] The tissue distribution in T-cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product in
T-cells indicates a role in the regulation of the proliferation;
survival; differentiation; and/or activation of potentially all
hematopoietic cell lineages, including blood stem cells. This gene
product may be involved in the regulation of cytokine production,
antigen presentation, or other processes that may also suggest a
usefulness in the treatment of cancer (e.g., by boosting immune
responses).
[0325] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immune deficiency diseases such as
AIDS, and leukemia. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tumors and tissues. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0326] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:52 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1241 of SEQ ID NO:52, b is an integer
of 15 to 1255, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:52, and where b is greater
than or equal to a+14.,
[0327] Features of Protein Encoded by Gene No: 43
[0328] The protein product of this gene was found to have homology
to the human CD84 protein which, as a novel member of the Ig
superfamily, is thought to play an important role in the modulation
of the immune response. The present invention appears to encode a
novel full-length CD84 homolog and is highly enriched, if not
specific, for activated T cells. In specific embodiments,
polypeptides of the invention comprise the following amino acid
sequence:
19 ITPLGLGAAD (SEQ ID NO: 442),
TLRVLGKVPAVCPWCALWRKAGMDMTYSWLSRGDSTYTFHEGPVLSTSWRPGDSALSYTCR (SEQ
ID NO: 443), ANNPISNVSSCPIPDGPFYADPNYASEKPSTAFCLLAKGLLIFLLLVILAM-
GLW VIRVQKRHKMPRMKKLMRNRMKLRKEAKPGSSPA AVCPWCALWRKAGMDMTYSWL (SEQ
ID NO: 444), PGDSALSYTCRANNPISNVSSCPI (SEQ ID NO: 445),
YASEKPSTAFCLLAKGLLIFLLLV (SEQ ID NO: 446), and/or
QKRHKMPRMKKLMRNRMKLRKEAKPG (SEQ ID NO: 447).
[0329] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0330] This gene is expressed primarily in human activated
T-Cells.
[0331] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immunodeficiencies, inflammatory conditions,
infections, and other immune or hematopoietic disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
disorders of the immune system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, hematopoietic, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0332] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 192 as residues: Glu-15 to Arg-23, Asn-79 to
Gly-84.
[0333] The tissue distribution in activated T-cells indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product in
T-cells indicates a role in the regulation of the proliferation;
survival; differentiation; and/or activation of potentially all
hematopoietic cell lineages, including blood stem cells. This gene
product may be involved in the regulation of cytokine production,
antigen presentation, or other processes that may also suggest a
usefulness in the treatment of cancer (e.g., by boosting immune
responses). Since the gene is expressed in cells of lymphoid
origin, the natural gene product may be involved in immune
functions. Therefore it may be also used as an agent for
immunological disorders including arthritis, asthma, immune
deficiency diseases such as AIDS, and leukemia. Protein, as well
as, antibodies directed against the protein may show utility as a
tumor marker and/or immunotherapy targets for the above listed
tumors and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0334] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID. NO:53 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1126 of SEQ ID NO:53, b is an integer
of 15 to 1140, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:53, and where b is greater
than or equal to a+14.
[0335] Features of Protein Encoded by Gene No: 44
[0336] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: IAWSGNIPSLLCLFEHDMSFQDE
(SEQ ID NO: 448). Polynucleotides encoding these polypeptides are
also encompassed by the invention.
[0337] This gene is expressed primarily in human tonsil.
[0338] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, inflammatory conditions, infections, or
immunodeficiencies, and immune or hematopoietic diseases and/or
disorders. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune diseases, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., immune, hematopoietic, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0339] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 193 as residues: Ile-2 to Lys-9, Gln-43 to Phe-49,
Asn-59 to His-69, Gly-87 to Asp-93.
[0340] The tissue distribution in tonsils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product
indicates a role in the regulation of the proliferation; survival;
differentiation; and/or activation of potentially all hematopoietic
cell lineages, including blood stem cells. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer (e.g., by boosting immune
responses).
[0341] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immune deficiency diseases such as
AIDS, and leukemia. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tumors and tissues. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0342] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:54 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1206 of SEQ ID NO:54, b is an integer
of 15 to 1220, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:54, and where b is greater
than or equal to a+14.
[0343] Features of Protein Encoded by Gene No: 45
[0344] The translation product of this gene shares sequence
homology with a novel human G52-24 secreted protein as well as the
early lymphocyte activation antigen CD69, the latter of which has
been shown to be important in lymphocyte proliferation and
functions as a signal trasmitting receptor in lymphocytes, natural
killer cells, and platelets (See Genseq and Genbank Accession Nos.
W27288 and gi.vertline.558352, respectively). Preferred
polypeptides comprise the following amino acid sequence:
20 ENFLLRYKGPSDHWIGLSREQGQPWKWINGTEWTRQLVMKEDGANLYVAKV (SEQ ID NO:
449), SQVPRMNPXLS WVLLCYPGWSAVXTIVAHCSLDFPGSK
ELTAIKSHQYVLQAACPESWIGFQRKCFYFSDDTKNWTSSQRFCDSQDADLAQ (SEQ ID NO:
450), VESFQELVRK WIGLSREQGQPWKWING (SEQ ID NO: 451), CPESWIGFQRKC
(SEQ ID NO: 452), NFLLRYKGPSDHWIGL (SEQ ID NO: 453),
ASHLRLLSSWDYRFPILGAGECAYLNDKGASSARHYTERKWI CSKSDIHV (SEQ ID NO:
454), ENFLLRYKGPSDHWIGLSREQGQPWKWINGTEWTRQLV (SEQ ID NO: 455),
MKEDGANLYVAKVSQVPRMNPXLS WVLLCYPGWSAVXTIVAHCSLDFPGSK
EQLEELELKKKDFIKILESVQGNWRQNEDSGKGPQRSCL (SEQ ID NO: 457),
FWPESKIQPYKDMFSCEII (SEQ ID NO: 458),
SWTSSLLNXCLHSKEHSIKATIWRLFFXILTIILCGMVAALSAIRANCHQ EPSVCSSSCMP (SEQ
ID NO: 456), RKLDWFSKKVFLFF EQLEELELKKKDFIKILESVQGNWRQ (SEQ ID NO:
459), and/or NEDSGKGPQRSCLHSKEHSIKATLIWRLFFLI
ENFLLRYKGPSDHWIGLXXEQGQP- WKWINGTEWTRQ (SEQ ID NO: 460).
[0345] Also preferred are the polynucleotides encoding these
polypeptides.
[0346] This gene is expressed primarily in human testes.
[0347] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive, endocrine, and/or immune or hematopoietic
disorders. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the disorders of reproductive system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, reproductive,
endcrine, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, seminal fluid, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0348] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 194 as residues: Asn-20 to Pro-25, Ser-48 to
Asp-65.
[0349] The tissue distribution in human testes indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of reproductive disorders, particularly autoimmune disorders,
infertility, or the protein may even be useful as a novel
contraceptive. Homology of this gene product to the early
lymphocyte activation antigen CD69 indicates a role in the
regulation of the proliferation; survival; differentiation; and/or
activation of potentially all hematopoietic cell lineages,
including blood stem cells. This gene product may be involved in
the regulation of cytokine production, antigen presentation, or
other processes that may also suggest a usefulness in the treatment
of cancer (e.g., by boosting immune responses).
[0350] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immune deficiency diseases such as
AIDS, and leukemia. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0351] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:55 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 680 of SEQ ID NO:55, b is an integer
of 15 to 694, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:55, and where b is greater
than or equal to a+14.
[0352] Features of Protein Encoded by Gene No: 46
[0353] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: RHEPDPM (SEQ ID NO:
461). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0354] This gene is expressed primarily in human testes.
[0355] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, male reproductive or endocrine disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., endocrine, reproductive, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, seminal fluid,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0356] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 195 as residues: Pro-20 to Trp-25, Arg-33 to Thr-38,
Asn-5 1 to Ile-56, Gly-82 to Ser-91, Lys-151 to Arg-156.
[0357] The tissue distribution in human testicular tissues and
cells indicates that polynucleotides and polypeptides corresponding
to this gene are useful for the detection, treatment, and/or
prevention of various endocrine disorders and cancers, particularly
Addison's disease, Cushing's Syndrome, and disorders and/or cancers
of the pancrease (e.g., diabetes mellitus), adrenal cortex,
ovaries, pituitary (e.g., hyper-, hypopituitarism), thyroid (e.g.,
hyper-, hypothyroidism), parathyroid (e.g., hyper-,
hypoparathyroidism), hypothallamus, and testes.
[0358] Alternatively, expression within the human testis may be
indicative for a role in normal testicular function, and may
implicate this gene product in male fertility, and could even
suggest its use as a novel contraceptive. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0359] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:56 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 974 of SEQ ID NO:56, b is an integer
of 15 to 988, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:56, and where b is greater
than or equal to a+14.
[0360] Features of Protein Encoded by Gene No: 47
[0361] One embodiment of this gene comprises polypeptides of the
following amino acid sequence:
21 LKGREAGAGPGTAGAPGREDANGXXRGRGGXHQLYLWVDNIPLSRPKRNLS (SEQ ID
NO:462), RDFSDGVLVAEVIKFYFPKMVEMHYVGTSSLQQKLSNWGHLNRKV- LKRL
NFSVPDDV WVDNIPLSRPKRNLSRDFSDGVLV- A (SEQ ID NO:463),
YVGTSSLQQKLSNWGHLNRKVLKRL (SEQ ID NO:464), GSAWRRG (SEQ ID NO:465),
RGAGSRAPAPYRSWLPRMAVATWMWVYPRRPEVKVSRTPREGVSSAGTG
RRRLGLQRITGRCRATPASSSRSLKRSRSCWPLKRPCRSCR WLPRMAVATWMWVYPRRPEVK
(SEQ ID NO:466), CRATPASSSRSLKRSRSCWPLKR (SEQ ID NO:467),
EHNTDFNGAALSRNLQTFRLSTPCARREGRLLRA (SEQ ID NO:468);
HRRCPPYSWRSHASPLPLQLLRSPSPRWVPGKLPGGAGEPLSGPGQIPPWLRA
WGTSLDGDAAVLGAGRGPDSGGVDRAKGPPPKAQRREMQGRAQGVGHCFG
GQARSLHVASGLWKAVHSPDPDLRSGRRRLSPGPALLEFLSHLLHAHPSQGRR
ALGPQQARESSGLRPPNGLSIGGWVRRGVGALAGTRASPRGPGRRSPLLTXR
XLEPPGEVFDPHILELEQVLQAPYLHLQDLHGLLRGQQLLLLFSDLEDEAGVA
LQRPVIRWRPRRRRPVPAELTPSLGVRDTFTSGLLGYTHIHVATAILGS QLL
TDFNGAALSRNLQTFRLSTPCARREG (SEQ ID NO:469),
RCPPYSWRSHASPLPLQLLRSPSPR (SEQ ID NO:470), GAGEPLSGPGQIPPWLRAWGTSLD
(SEQ ID NO:471), LGAGRGPDSGGVDRAKGPPPKAQRREMQGR (SEQ ID NO:472),
QARSLHVASGLWKAVHSPDPDLR (SEQ ID NO:473), HPSQGRRALGPQQARESSGL (SEQ
ID NO:474), IGGWVRRGVGALAGTRASPRGPGRRSP (SEQ ID NO: 475),
EPPGEVFDPHILELEQVLQAPYLHL (SEQ ID NO:476), and/or
VPAELTPSLGVRDTFTSGLLGYTHIHVA (SEQ ID NO:477).
[0362] An additional embodiment is the polynucleotides encoding
these polypeptides.
[0363] This gene is expressed primarily in human adult testis.
[0364] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, reproductive and/or endocrine disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
disorders of the reproductive system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., reproductive, endocrine,
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
seminal fluid, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0365] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 196 as residues: Gln-21 to Gly-33, Gln-55 to
Glu-60.
[0366] The tissue distribution in testicular tissue indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection, treatment, and/or prevention of
reproductive system disorders, and may be indicative of a role for
this gene product in normal testicular function, male fertility,
and/or as a male contraceptive. Protein, as well as, antibodies
directed against the protein may show utility as a tissue-specific
marker and/or immunotherapy target for the above listed tissues.
.
[0367] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:57 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1486 of SEQ ID NO:57, b is an integer
of 15 to 1500, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:57, and where b is greater
than or equal to a+14.
[0368] Features of Protein Encoded by Gene No: 48
[0369] The translation product of this gene shares sequence
homology with the human M phase phosphoprotein 10 as well as ORF
YJR002w of Saccharomyces cerevisiae (See Genbank Accession
No.gnl.vertline.PID.vertl- ine.e266673) which are thought to play
important roles in the regulation of cellular division. Preferred
polypeptides comprise the following amino acid sequence:
22 AKNSQKEENPEHVEIQKMMDSLFLKLDALSNFHFIPKPPVEIKVVSNLPAI (SEQ ID
NO:478), TMEEVAPVSVSDAALLAPEEIKEKNKAGDIKTAAEKTATDKKRERRK- KKYQKR
MKIKEKEKRRKLLEKSSVDQAGKYSKTVASEKLKQLTKTGKASFIKVRTR
ERKLLKGTFVGEVDSKCWVTGMSEPADSPPVG
LQDEGKDKALKSSQAFFSKLQDQVKMQINDAKKTEKKKKKRQDISVHKLKL (SEQ ID
NO:479), DEGKDKALKSSQAFFSKLQDQVKMQINDA (SEQ ID NO:480),
EENPEHVEIQKMMDSLFLKLDALSNFHF (SEQ ID NO:481),
SSVDQAGKYSKTVASEKLKQLTKTGKASFIK (SEQ ID NO:483),
VSVSDAALLAPEEIKEKNKAGDI (SEQ ID NO:484), VLEVMVTVAPK (SEQ ID
NO:485), LQDEGKDKALKSSQAFFSKLQDQVKMQINDAKKTE (SEQ ID NO:486),
[0370] Also preferred are the polynucleotides encoding these
polypeptides.
[0371] This gene is expressed primarily in human thyroid.
-Therefore, polynucleotides and polypeptides of the invention are
useful as reagents for differential identification of the tissue(s)
or cell type(s) present in a biological sample and for diagnosis of
diseases and conditions which include, but are not limited to,
endocrine, proliferative, or developmental disorders, particularly
diseases relating to the thyroid gland, particularly hyper- and
hypothyroidism. Similarly, polypeptides and antibodies directed to
these polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the disorders of the endocrine system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., endocrine, developmental,
metabolic, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.:,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0372] The tissue distribution in human thyroid indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for metabolic disorders, particularly hyper-,
hypothyroidism, Graves' disease, Hashimoto's thyroiditis, and/or
cancer or neoplasias of the thyroid, and/or other endocrine organs
and immune system. Moreover, the protein may show utility in the
diagnosis, prevention, and/or treatment of developmental disorders.
In addition, the homology to an M phase phosphoprotein indicates it
may be a key player in the proliferation, maintenance, and/or
differentiation of various cell types during development. It may
also act as a morphogen to control cell and tissue type
specification. Because of potential roles in proliferation and
differentiation, this gene product may have applications in the
adult for tissue regeneration and the treatment of cancers.
Protein, as well as, antibodies directed against the protein may
show utility as a tissue-specific marker and/or immunotherapy
target for the above listed tissues.
[0373] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:58 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1377 of SEQ ID NO:58, b is an integer
of 15 to 1391, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:58, and where b is greater
than or equal to a+14.
[0374] Features of Protein Encoded by Gene No: 49
[0375] The translation product of this gene was found to have
homology to the cell division control protein 48 (cdc48) of
Methanococcus jannaschii (See Genbank Accession
No.gi.vertline.1591785) which is thought to play a key role in the
regulation of cellular division. In specific embodiments,
polypeptides of the invention comprise the following amino acid
sequence:
23 HEAAQGAVCRGQGAPATNPQAPVAAAARVARRVN (SEQ ID NO:487), KIPS (SEQ ID
NO:488), ANRRATRCLGCDHQNFVKVRNKHKGKPTFME- EVLEHLPGKTQDEVQQHEKW
YQKFLALEERKKESIQIWKTKKQQKREEIFKLKEKAD- NTPVLFHNKQEDNQKQ
KEEQRKKQKLAVEAWKKQKSIEMSMKCASQLKKKKKKKKKN- QKERQRQFK
LKLLLESYTQQKKEQEEFLRLEKEIREKAEKAEKRKNAADEISRFQER- DLHKLE
LKILDRQAKEDEKSQKQRRLAKLKEKVENNVSRDPSRLYKPTK
VKVRNKHKGKPTFMEEVLEHLPGK (SEQ ID NO:489), QHEKWYQKFLALEERKKESIQIW
(SEQ ID NO:490), FKLKEKADNTPVLFHNKQEDNQKQKEEQRKK (SEQ ID NO:491),
FLRLEKIEIREKAEKAEKRKNAADEISRFQERDLHKL (SEQ ID NO:492), and/or
KQRRLAKLKEKVENNVSRDPSRLY (SEQ ID NO:493).
[0376] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0377] This gene is expressed primarily in pancreas, and to a
lesser extent in kidney and bone marrow.
[0378] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, pancreas, urogenital, developmental, metabolic, immune,
and/or hematopoietic disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification, of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the pancreas, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., endocrine,
developmental, metabolic, immune, hematopoietic, gastrointestinal,
and cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, bile, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0379] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO:198 as residues: Pro-35 to Cys-43, Gln-56 to Lys-67,
Thr-73 to Lys-78, Tyr-93 to Asp-98, Ser-116 to Gln-125, Leu-142 to
Phe-151, Phe-169 to Arg-174, Ile-181 to Glu-190, Thr-243 to
Gly-248.
[0380] The tissue distribution in pancreas indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection, treatment, and/or prevention of various
endocrine disorders and cancers, particularly Addison's disease,
Cushing's Syndrome, and disorders and/or cancers of the pancrease
(e.g., diabetes mellitus), adrenal cortex, ovaries, pituitary
(e.g., hyper-, hypopituitarism), thyroid (e.g., hyper-,
hypothyroidism), parathyroid (e.g., hyper-,hypoparathyroidism),
hypothallamus, and testes. Alternatively, the expression within
bone marrow indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the treatment
and-diagnosis of hematopoetic related disorders such as anemia,
pancytopenia, leukopenia, thrombocytopenia or leukemia since
stromal cells are important in the production of cells of
hematopoietic lineages. The uses include bone marrow cell ex vivo
culture, bone marrow transplantation, bone marrow reconstitution,
radiotherapy or chemotherapy of neoplasia. The gene product may
also be involved in lymphopoiesis, therefore, it can be used in
immune disorders such as infection, inflammation, allergy,
immunodeficiency etc. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types.
[0381] Moreover, the protein product of this gene could be used in
the treatment and/or detection of kidney diseases including renal
failure, nephritus, renal tubular acidosis, proteinuria, pyuria,
edema, pyelonephritis, hydronephritis, nephrotic syndrome, crush
syndrome, glomerulonephritis, hematuria, renal colic and kidney
stones, in addition to Wilm's Tumor Disease, and congenital kidney
abnormalities such as horseshoe kidney, polycystic kidney, and
Falconi's syndrome. Considering the homology to a conserved cell
division control protein indicates that the protein may show
utility in the diagnosis, prevention, and/or treatment of
developmental disorders, and may even serve as a suppressor in
tumorigenesis. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0382] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:59 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1565 of SEQ ID NO:59, b is an integer
of 15 to 1579, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:59, and where b is greater
than or equal to a+14.
[0383] Features of Protein Encoded by Gene No: 50
[0384] The translation product of this gene was shown to have
homology to the chicken LRP/alpha-2-macroglobulin receptor which is
thought to play a pivitol role on the metabolism of
alpha-2-macroglubulins, as well as, complexes between plasminogen
activators and their endogenous inhibitors (See Genbank Accession
No.gb.vertline.X74904.vertline.GGLRPA2MR).
[0385] This gene is expressed primarily in neuronal tissues, and to
a lesser extent in uterine cancer.
[0386] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neuronal disorders and uterine cancer. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s) For a number of
disorders of the above tissues or cells, particularly of the
central neuron system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., neural, reproductive, cancerous and wounded
tissues) or bodily fluids (e.g., amniotic fluid, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0387] The tissue distribution in neuronal tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses,
autism, and altered behaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing embryo,
sexually-linked disorders, or disorders of the cardiovascular
system. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0388] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:60 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1227 of SEQ ID NO:60, b is an integer
of 15 to 1241, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:60, and where b is greater
than or equal to a+14.
[0389] Features of Protein Encoded by Gene No: 51
[0390] This gene is expressed primarily in uterine cancer.
[0391] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, uterine cancer, and other reproductive disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the uterine cancer, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types (e.g., reproductive, cancerous and wounded tissues) or
bodily fluids (e.g., amniotic fluid, serum, plasma, urine, synovial
fluid and spinal fluid) or another tissue or cell sample taken from
an individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0392] The tissue distribution in tumors of the uterus indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for diagnosis and intervention of these tumors or
proliferative conditions, in addition to other tumors or cell
types. Protein, as well as, antibodies directed against the protein
may show utility as a tissue-specific marker and/or immunotherapy
target for the above listed tissues.
[0393] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:61 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 916 of SEQ ID NO:61, b is an integer
of 15 to 930, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:61, and where b is greater
than or equal to a+14.
[0394] Features of Protein Encoded by Gene No: 52
[0395] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: VKPPDQSCNHWRDEQCLV (SEQ
ID NO: 494). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0396] This gene is expressed primarily in wilm's tumor.
[0397] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, Wilm's tumor, and other urogenital disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the Wilm's tumor, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types (e.g., urogenital, cancerous and wounded tissues) or
bodily fluids (e.g., serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0398] The tissue distribution in Wilm's tumor indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis and treatment of Wilm's tumor. Protein, as
well as, antibodies directed against the protein may show utility
as a tissue-specific marker and/or immunotherapy target for the
above listed tissues. Furthermore, this gene or gene product is
useful in the treatment and/or detection of kidney diseases
including renal failure, nephritus, renal tubular acidosis,
proteinuria, pyuria, edema, pyelonephritis, hydronephritis,
nephrotic syndrome, crush syndrome, glomerulonephritis, hematuria,
renal colic and kidney stones, in addition to congenital kidney
abnormalities such as horseshoe kidney, polycystic kidney, and
Falconi's syndrome. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0399] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:62 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 984 of SEQ ID NO:62, b is an integer
of 15 to 998, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:62, and where b is greater
than or equal to a+14.
[0400] Features of Protein Encoded by Gene No: 53
[0401] The translation product of this gene was shown to have
homology to the MEK kinase 3 of Mus musculus, mutations of which
and/or aberrant regulation of, may provide a predisposition to
cancer. The gene encoding the disclosed cDNA is thought to reside
on chromosome 17. Accordingly, polynucleotides related to this
invention are useful as a marker in linkage analysis for chromosome
17.
[0402] This gene is expressed primarily in pituitary, and to a
lesser extent in ulcerative colitis and hematopoietic cells.
[0403] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune, gastrointestinal, hematopoietic diseases.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the neuronal and immune tissues, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., neuronal, immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0404] The tissue distribution in hematopoietic tissues indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for the diagnosis, treatment, and/or prevention of a
variety of immune system disorders. Expression of this gene product
in ulcerative colitis indicates a role in the regulation of the
proliferation; survival; differentiation; and/or activation of
potentially all hematopoietic cell lineages, including blood stem
cells. This gene product may be involved in the regulation of
cytokine production, antigen presentation, or other processes that
may also suggest a usefulness in the treatment of cancer (e.g., by
boosting immune responses). Since the gene is expressed in cells of
lymphoid origin, the natural gene product may be involved in immune
functions. Therefore it may be also used as an agent for
immunological disorders including arthritis, asthma, immune
deficiency diseases such as AIDS, and leukemia. Protein, as well
as, antibodies directed against the protein may show utility as a
tumor marker and/or immunotherapy targets for the above listed
tumors and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types.
[0405] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:63 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1179 of SEQ ID NO:63, b is an integer
of 15 to 1193, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:63, and where b is greater
than or equal to a+14.
[0406] Features of Protein Encoded by Gene No: 54
[0407] When tested against Jurkat T-cell cell lines, supernatants
removed from cells containing this gene activated the GAS (gamma
activation site) pathway. Thus, it is likely that this gene
activates T-cells through the Jaks-STAT signal transduction
pathway. GAS (gamma activation site) is a promoter element found
upstream in many genes which are involved in the Jaks-STAT pathway.
The Jaks-STAT pathway is a large, signal transduction pathway
involved in the differentiation and proliferation of cells.
Therefore, activation of the Jaks-STATs pathway, reflected by the
binding of the GAS element, can be used to indicate proteins
involved in the proliferation and differentiation of cells.
[0408] This gene is expressed primarily in fetal spleen and adipose
tissues.
[0409] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune, metabolic, and developmental disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the fetal spleen and adipose tissues, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, developing, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0410] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 203 as residues: Tyr-41 to Phe-47.
[0411] The tissue distribution in fetal liver/spleen, combined with
the detection of GAS promoter activation activity, indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of a variety
of immune system disorders. Expression of this gene product in
fetal spleen indicates a role in the regulation of the
proliferation; survival; differentiation; and/or activation of
potentially all hematopoietic cell lineages, including blood stem
cells. This gene product may be involved in the regulation of
cytokine production, antigen presentation, or other processes that
may also suggest a usefulness in the treatment of cancer (e.g., by
boosting immune responses).
[0412] Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immune deficiency diseases such as
AIDS, and leukemia. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tumors and tissues. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0413] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:64 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 816 of SEQ ID NO:64, b is an integer
of 15 to 830, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:64, and where b is greater
than or equal to a+14.
[0414] Features of Protein Encoded by Gene No: 55
[0415] This gene is expressed primarily in IL-1/TNF stimulated
synovial and human adipose tissues.
[0416] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, rheumatoid arthritis or obessity, and disorders of the
musculo-skeletal system. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune and musculo-skeletal
systems, expression of this gene at significantly higher or lower
levels may. be routinely detected in certain tissues or cell types
and cell types (e.g., synovial and adipose cells and tissues,
musculo-skeletal, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0417] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 204 as residues: Leu-37 to Arg-45, Ser-60 to
Ser-65.
[0418] The tissue distribution in synovial tissue and adipose
tissue indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the diagnosis or
treatment of rheumatoid arthritis or other immune diseases. In
addition, the expression of this gene product in synovium indicates
a role in the detection and treatment of disorders and conditions
affecting the skeletal system, in particular osteoporosis as well
as disorders afflicting connective tissues (e.g. arthritis, trauma,
tendonitis, chrondomalacia and inflammation), such as in the
diagnosis or treatment of various autoimmune disorders such as
rheumatoid arthritis, lupus, scleroderma, and dermatomyositis as
well as dwarfism, spinal deformation, and specific joint
abnormalities as well as chondrodysplasias (ie. spondyloepiphyseal
dysplasia congenita, familial osteoarthritis, Atelosteogenesis type
II, metaphyseal chondrodysplasia type Schmid).
[0419] The tissue distribution in adipose tissue indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment of obesity and other metabolic and
endocrine conditions or disorders. Furthermore, the protein product
of this gene may show utility in ameliorating conditions which
occur secondary to aberrant fatty-acid metabolism (e.g. aberrant
myelin sheath development), either directly or indirectly. Protein,
as well as, antibodies directed against the protein may show
utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0420] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:65 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 853 of SEQ ID NO:65, b is an integer
of 15 to 867, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:65, and where b is greater
than or equal to a+14.
[0421] Features of Protein Encoded by Gene No: 56
[0422] When tested against K562 leukemia cell lines, supernatants
removed from cells containing this gene activated the ISRE assay.
Thus, it is likely that this gene activates leukemia cells through
the Jak-STAT signal transduction pathway. The interferon-sensitive
response element is a promoter element found upstream of many genes
which are involved in the Jak-STAT pathway. The Jak-STAT pathway is
a large, signal transduction pathway involved in the
differentiation and proliferation of cells. Therefore, activation
of the Jak-STAT pathway, reflected by the binding of the ISRE
element, can be used to indicate proteins involved in the
proliferation and differentiation of cells.
[0423] This gene is expressed primarily in aortic endothelium, and
to a lesser extent in melanocyte.
[0424] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, cardiovascular diseases. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the cardiovascular system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
vascular, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0425] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 205 as residues: Met-1 to Trp-12, Arg-33 to
Ser-53.
[0426] The tissue distribution in human aortic endothelial cells
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the detection or intervention of
cardiovascular diseases, such as hypertension, cadiovascular
injuries, congenital heart diseases, ischemic heart diseases,
rheumatic and other hypersensitivity diseases, cardiomyopathy,
restenosis, atherosclerosis, stoke, angina, thrombosis, and wound
healing. Protein, as well as, antibodies directed against the
protein may show utility as a tissue-specific marker and/or
immunotherapy target for the above listed tissues.
[0427] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:66 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 671 of SEQ ID NO:66, b is an integer
of 15 to 685, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:66, and where b is greater
than or equal to a+14.
[0428] Features of Protein Encoded by Gene No: 57
[0429] The translation product of this gene shares sequence
homology with prostaglandin EP3-9 receptor, which is thought to be
important in prostaglandin hormonal reaction. In specific
embodiments, polypeptides of the invention comprise the following
amino acid sequence: MAIPAFSSCQQISSAAALQI (SEQ ID NO: 495), and/or
CNGPFKHFSFTVST (SEQ ID NO: 496). Polynucleotides encoding these
polypeptides are also encompassed by the invention.
[0430] This gene is expressed primarily in human retina.
[0431] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, glaucoma or other ocular diseases. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the ocular
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., retinal and other optic tissue, tissue of the nervous
system, and cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0432] The tissue distribution in retinal tissues and the homology
to prostaglandin receptor indicates that polynucleotides and
polypeptides corresponding to this gene are useful for the
detection and intervention of ocular diseases like glaucoma.
Specifically, the receptor can be used for the identification of
agonists or antagonists, anti-inflammatories for the eyes, and
vasoconstrictive agents, etc. Furthermore, the tissue distribution
in retina indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the treatment and/or
detection of eye disorders including blindness, color blindness,
impaired vision, short and long sightedness, retinitis pigmentosa,
retinitis proliferans, and retinoblastoma.
[0433] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:67 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 787 of SEQ ID NO:67, b is an integer
of 15 to 801, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:67, and where b is greater
than or equal to a+14.
[0434] Features of Protein Encoded by Gene No: 58
[0435] The translation product of this gene shares weak sequence
homology with Hemophilus influenzae outmembrane protein P6 which is
thought to be important in host cell interaction.
[0436] This gene is expressed primarily in human adrenal gland
tumor.
[0437] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, adrenal insufficiency or hyperfunction, adrenal gland
tumors. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the endocrine systems and cancers thereof, expression of this gene
at significantly higher or lower levels may be routinely detected
in certain tissues or cell types (e.g., adrenal gland, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0438] The tissue distribution in adrenal gland tumor and homology
to Haemophilus influenzae outer membrane protein suggest that
polynucleotides and polypeptides corresponding to this gene are
useful for adrenal insufficiencies or hyperfunction, because a
secretory protein from an endocrine organ may function as a
hormone. The protein product of this gene is also useful as a
diagnostic and/or treatment for adrenal gland tumors, as well as
tumors of other tissues where expression has been observed.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0439] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:68 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 894 of SEQ ID NO:68, b is an integer
of 15 to 908, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:68, and where b is greater
than or equal to a+14.
[0440] Features of Protein Encoded by Gene No: 59
[0441] When tested against a Jurkat T-cell line,
supernatants-removed from cells containing this gene activated the
GAS (gamma activation site) pathway. Thus, it is likely that this
gene activates T-cells through the Jaks-STAT signal transduction
pathway. The GAS is a promoter element found upstream in many genes
which are involved in the Jaks-STAT pathway. The Jaks-STAT pathway
is a complex, signal transduction pathway involved in the
differentiation and proliferation of cells. Therefore, activation
of the Jaks-STATs pathway, reflected by the binding of the GAS
element, can be used to indicate proteins involved in the
proliferation and differentiation of cells.
[0442] This gene is expressed primarily in human kidney pyramid,
and to a lesser extent in human brain.
[0443] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, nephrotic, nephritic syndromes, renal failure,
hypertensive nephrosclerosis. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the renal system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., renal, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0444] The tissue distribution in kidney indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for renal diseases, including renal failure, nephritus,
renal tubular acidosis, proteinuria, pyuria, edema, pyelonephritis,
hydronephritis, nephrotic syndrome, crush syndrome,
glomerulonephritis, hematuria, renal colic and kidney stones, in
addition to Wilm's Tumor Disease, and congenital kidney
abnormalities such as horseshoe kidney, polycystic kidney, and
Falconi's syndrome. Additionally, the gene product may have
endocrine functions related to renal function, metabolism and
homeostasis. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0445] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:69 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 682 of SEQ ID NO:69, b is an integer
of 15 to 696, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:69, and where b is greater
than or equal to a+14.
[0446] Features of Protein Encoded by Gene No: 60
[0447] This gene is expressed primarily in both normal or cancerous
human breast tissue.
[0448] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, Non-neoplastic breast diseases or breast cancers.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the breast, expression of this gene at significantly higher or
lower levels may be routinely detected in certain tissues or cell
types (e.g., mammary tissue, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0449] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 209 as residues: Pro-20 to Ser-28.
[0450] The tissue distribution in breast indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for either non-neoplastic breast diseases, such as
congentital anomalities, gynecomastia, mastitis and abscess, duct
ectasia and fat necrosis, or neoplasia in the breast. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0451] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:70 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 441 of SEQ ID NO:70, b is an integer
of 15 to 455, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:70, and where b is greater
than or equal to a+14.
[0452] Features of Protein Encoded by Gene No: 61
[0453] When tested against a K562 cell line, supernatants removed
from cells containing this gene activated the ISRE
(interferon-sensitive responsive element) pathway. Thus, it is
likely that this gene activates leukemia cells, or more generally,
immunr or hematopoietic cells, or other cells or cell-types,
through the Jaks-STAT Signal transduction pathway. The ISRE is a
promoter element found upstream in many genes which are involved in
the Jaks-STAT pathway. The Jaks-STAT pathway is a complex, signal
transduction pathway involved in the differentiation and
proliferation of cells. Therefore, activation of the Jaks-STATs
pathway, reflected by the binding of the ISRE element, can be used
to indicate proteins involved in the proliferation and
differentiation of cells. In specific embodiments, polypeptides of
the invention comprise the following amino acid sequence:
IRHERLWAELALLTGRNE (SEQ ID NO: 497). Polynucleotides encoding these
polypeptides are also encompassed by the invention. The gene
encoding the disclosed cDNA is thought to reside on chromosome 3.
Accordingly, polynucleotides related to this invention are useful
as a marker in linkage analysis for chromosome 3.
[0454] This gene is expressed primarily in activated T-cells and
osteoarthritis, and to a lesser extent in aortic endothelium and
placenta.
[0455] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, inflammatory conditions, vascular disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
system and vascular tissues, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types and cell types (e.g., T-cells and
other cells and tissue of the immune system, bone tissue,
endothelium and placenta, vascular tissue, and cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0456] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 210 as residues: Gln-36 to Glu-49, Glu-51 to Leu-66,
Asp-68 to Ser-73.
[0457] The tissue distribution in activated T-cells and under
inflammatory conditions like osteoarthritis suggest that the
protein product of this gene is involved in the inflammatory
reactions. Therefore it may be useful in the diagnosis or
intervention in the inflammatory diseases with the involvement of
T-cells, including osteoarthritis. Furthermore, the tissue
distribution indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the diagnosis and/or
treatment of disorders of the placenta. Specific expression within
the placenta indicates that this gene product may play a role in
the proper establishment and maintenance of placental function.
[0458] Alternately, this gene product may be produced by the
placenta and then transported to the embryo, where it may play a
crucial role in the development and/or survival of the developing
embryo or fetus. Expression of this gene product in a vascular-rich
tissue such as the placenta also indicates that this gene product
may be produced more generally in endothelial cells or within the
circulation. In such instances, it may play more generalized roles
in vascular function, such as in angiogenesis. It may also be
produced in the vasculature and have effects on other cells within
the circulation, such as hematopoietic cells. It may serve to
promote the proliferation, survival, activation, and/or
differentiation of hematopoietic cells, as well as other cells
throughout the body.
[0459] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:71 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 399 of SEQ ID NO:71, b is an integer
of 15 to 413, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:71, and where b is greater
than or equal to a+14.
[0460] Features of Protein Encoded by Gene No: 62
[0461] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
24 GTESPMVMCCREVSQSENCLFLDTTFRFIFGKTFTNHDYISIHFYFLKAFLFSFFYSNV (SEQ
ID NO:498).
[0462] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0463] This gene is expressed primarily in breast lymph nodes,
B-cell lymphoma, and to a lesser extent in neutrophils and bone
marrow cells.
[0464] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, inflammation, immunodeficiency, allergy. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
system, expression of this gene at significantly higher or lower
levels may-be routinely detected in certain tissues or cell types
and cell types (e.g., blood cells, hematopoietic cells, and cells
and tissue of the immune system, cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0465] The tissue distribution in the cells of immunological
functions indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the diagnosis or
intervention of immunologically mediated disorders, such as
allergy, immunodeficiency, immune surveillance, etc. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0466] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:72 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 835 of SEQ ID NO:72, b is an integer
of 15 to 849, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:72, and where b is greater
than or equal to a+14.
[0467] Features of Protein Encoded by Gene No: 63
[0468] The translation product of this gene shares weak sequence
homology with Interferon induced 1-8 gene encoded polypeptide,
which is thought to be important in retroviral REV responsive
element binding and thus viral replication.
[0469] This gene is expressed primarily in B-cell lymphoma.
[0470] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune response to viral infections and other
immunologically related disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types and cell types (e.g.,
T-cells and other cells and tissue of the immune system, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0471] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 212 as residues: Pro-47 to Asn-53.
[0472] The tissue distribution in B-cell lymphoma and homology to
interferon induced 1-8 gene indicates that polynucleotides and
polypeptides corresponding to this gene are useful for the
intervention of viral infection and other immunologically related
disorders. The homology with interferon induced 1-8 REV response
element binding gene indicates the gene product may bind to viral
components to interfere with the entry, packaging, replication, or
induce the host cell anti-viral response by intereferon mediated
pathways. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0473] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:73 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between I to 491 of SEQ ID NO:73, b is an integer
of 15 to 505, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:73, and where b is greater
than or equal to a+14.
[0474] Features of Protein Encoded by Gene No: 64
[0475] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: IRHEEKGGKAQRWAE (SEQ ID
NO: 499). Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0476] This gene is expressed primarily in bone marrow.
[0477] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, hemapoiesis disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the hemapoietic system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g., bone
marrow, and cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0478] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 23 as residues: Thr-45 to Tyr-50.
[0479] The tissue distribution in bone marrow indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for hemapoiesis disorders. The gene product may function as
a growth factor or mobilization agent for the cells of myeloid or
lymphoid lineages. Furthermore, the polypeptides or polynucleotides
are also useful to enhance or protect proliferation,
differentiation, and functional activation of hematopoietic
progenitor cells (e.g., bone marrow cells), useful in treating
cancer patients undergoing chemotherapy or patients undergoing bone
marrow transplantation. Protein, as well as, antibodies directed
against the protein may show utility as a tissue-specific marker
and/or immunotherapy target for the above listed tissues.
[0480] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:74 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 705 of SEQ ID NO:74, b is an integer
of 15 to 719, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:74, and where b is greater
than or equal to a+14.
[0481] Features of Protein Encoded by Gene No: 65
[0482] The translation product of this gene shares sequence
homology familial adenomatous polyposis gene which is thought to be
important in the tumorigenesis of colon cancer (see, e.g., Fulton,
Nature 368, 32-38 (1994); accession no. U28412; Joslyn et al., Cell
66 (3), 601-613 (1991); accession no. M73547; and Spirio et al.,
Nucleic Acids Res. 19 (22), 6348 (1991)). In specific embodiments,
polypeptides of the invention comprise the following amino acid
sequence:
25 CRWRPESAAPC (SEQ ID NO:500), TRPGRGAQAPVK (SEQ ID NO:501),
MVSWMISRAVVLVFGMLYPAY (SEQ ID NO:502), GMLYPAYYSYKAVKTKN (SEQ ID
NO:503), EYVRWMMYWIVFALYTV (SEQ ID NO:504), YPAYYSYKAVKTKNVKE (SEQ
ID NO:505), VAWFPLYYELKIA (SEQ ID NO:506), and/or
MVSWMISRAVVLVFGMLYPAYYSYK (SEQ ID NO:507).
AVKTKNVKEYVRWMMYWIVFALYTVIETVADQTVAWFPLYYELKIAFVIWLLS
PYTKGASLIYRKFLHPLLSSKEREIDDYIVQAKERGYETMVNFGRQGLNLAATA
AVTAAVKSQGAITERLRSFSMHDLTTIQGDEPVGQRPYQPLPEAKKKSXQPPVN
QXVMEFHXKTXMXKQXKKQRGHIQLMRC
[0483] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0484] This gene is expressed primarily in osteoclastoma, prostate,
bone marrow and to a lesser extent in testes and dendritic cells.
Northern data has demonstrated that an abundant 1.3 kb band is seen
in testes tissues.
[0485] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, colon cancer and cancers of various origin, including
osteoclastoma and prostate cancer, as well as reproductive
disorders. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the tumorigenesis and reproductive disorders, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types and cell types (e.g.,
bone, prostate, reproductive, bone marrow, colon and other
gastrointestinal tissue, tissue of the nervous system, and testis
and other reproductive tissue, and cancerous and wounded tissues)
or bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0486] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 214 as residues: Ser-59 to Ile-64, Ala-71 to Tyr-76,
Pro-125 to Ser-141.
[0487] The tissue distribution in osteoclastoma, prostate, bone
marrow and homology to familial adenomatous polyposis gene
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for diagnosis and intervention of tumors of
various origins, including colon cancer, osteoclastoma and prostate
cancer. Alternatively, the Northern data demonstrating expression
in testes tissues indicates that the translation product of this
gene is useful for the diagnosis and/or treatment of reproductive
disorders and conditions concerning proper testicular function
(e.g., endocrine function, sperm maturation), as well as cancer.
Therefore, this gene product is useful in the treatment of male
infertility and/or impotence.
[0488] This gene product is also useful in assays designed to
identify binding agents, as such agents (antagonists) are useful as
male contraceptive agents. Similarly, the protein is believed to be
useful in the treatment and/or diagnosis of testicular cancer. The
testes are also a site of active gene expression of transcripts
that may be expressed, particularly at low levels, in other tissues
of the body. Therefore, this gene product may be expressed in other
specific tissues or organs where it may play related functional
roles in other processes, such as hematopoiesis, inflammation, bone
formation, and kidney function, to name a few possible target
indications.
[0489] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:75 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence is cumbersome. Accordingly, preferably excluded from the
present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1260 of SEQ ID NO:75, b is an integer
of 15 to 1274, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:75, and where the b is
greater than or equal to a+14.
[0490] Features of Protein Encoded by Gene No: 66
[0491] The translation product of this gene shares regional and
weak sequence homology with neu differentiation factor and a serine
protease N-terminal fragment which contains a EGF-like domain and
is thought to be important in the growth and differentiation of
several cell types, including colon epithelial cells and Schwann
cells.
[0492] This gene is expressed primarily in fetal lung, bone marrow,
fetal liver, and to a lesser extent in brain.
[0493] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, tissue injuries or diseases in lung, bone marrow, or
liver. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the liver and lung, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types and cell types (e.g., lung and pulmonary tissue, bone
marrow, hepatic tissue, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0494] The tissue distribution in fetal liver, combined with the
homology to neu differentiation factor indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis or intervention of liver or lung injuries,
including hepatic failure, recovery from hepatitis, cirrhosis,
hepatoblastoma, jaundice, liver metabolic diseases and conditions
that are attributable to the differentiation of hepatocyte
progenitor cells, and complications from liver transplantation.
[0495] Moreover, the protein product of this clone is useful for
the treatment and diagnosis of hematopoietic related disorders such
as anemia, pancytopenia, leukopenia, thrombocytopenia or leukemia
since stromal cells are important in the production of cells of
hematopoietic lineages The uses include bone marrow cell ex-vivo
culture, bone marrow transplantation, bone marrow reconstitution,
radiotherapy or chemotherapy of neoplasia. The gene product may
also be involved in lymphopoiesis, therefore, it can be used in
immune disorders such as infection, inflammation, allergy,
immunodeficiency etc. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0496] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:76 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 505 of SEQ ID NO:76, b is an integer
of 15 to 519, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:76, and where b is greater
than or equal to a+14.
[0497] Features of Protein Encoded by Gene No: 67
[0498] This gene is expressed primarily in activated T-cells.
[0499] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, arthritis, asthma, auto-immune and immunodeficiency
diseases. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune system, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types and cell types (e.g., T-cells and other cells and tissue
of the immune system, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0500] The expression of this gene in T-cells indicates a potential
role in the treament/detection of immune disorders such as
arthritis, asthma, hypersensitivity reactions and transplant
rejection, and also in immune deficiency diseases such as AIDS, and
leukemia. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0501] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:77 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 375 of SEQ ID NO:77, b is an integer
of 15 to 389, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:77, and where b is greater
than or equal to a+14.
[0502] Features of Protein Encoded by Gene No: 68
[0503] The gene encoding the disclosed cDNA is thought to reside on
chromosome 7. Accordingly, polynucleotides related to this
invention are useful as a marker in linkage analysis for chromosome
7.
[0504] This gene is expressed primarily in brain, and to a lesser
extent in breast.
[0505] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neurodegenerative conditions and behavioural disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., brain and other tissue of the
nervous system, mammary tissue, and cancerous and wounded tissues)
or bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0506] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 217 as residues: Leu-40 to His-46.
[0507] The tissue distribution in brain indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, schizophrenia, mania,
dementia, paranoia, obsessive compulsive disorder and panic
disorder. In addition, the gene or gene product may also play a
role in the treatment and/or detection of developmental disorders
associated with the developing embryo, or sexually-linked
disorders. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0508] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:78 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 809 of SEQ ID NO:78, b is an integer
of 15 to 823, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:78, and where b is greater
than or equal to a+14.
[0509] Features of Protein Encoded by Gene No: 69
[0510] The translation product of this gene shares sequence
homology with a rat secretory carrier membrane protein which is
believed to play a role in cell surface re-cycling. See e.g., Brand
et al., EMBO J 1993 October;12(10):3753-3761. Secretory carrier
membrane proteins (SCAMPs) are widely distributed as components of
post-Golgi membranes that function as recycling carriers to the
cell surface. In fibroblasts, SCAMPs are concentrated in
compartments involved in the endocytosis and recycling of cell
surface receptors while in neurons and other cell types having
regulated transport pathways, SCAMPs are also components of
regulated carriers (synaptic vesicles, secretion granules and
transporter vesicles). Their presence in multiple pathways
distinguishes them from proteins (e.g., recycling cell surface
receptors and synaptic vesicle proteins) which are concentrated in
selected pathways. The SCAMPs also do not appear to reside beyond
the boundaries of these pathways. This distribution indicates that
SCAMPs are general markers of membranes that function in cell
surface recycling. Accordingly, polpeptides of the invention and
antibodies thereto, may be used to identify membranes that function
in cell surface recycling. The gene encoding the disclosed cDNA is
thought to reside on chromosome 15. Accordingly, polynucleotides
related to this invention are useful as a marker in linkage
analysis for chromosome 15.
[0511] This gene is expressed primarily in hematopoietic cell
types.
[0512] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune and hematopoetic disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
and hematopoetic systems, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types and cell types (e.g., hematopoietic cells, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0513] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 218 as residues: Ser-25 to Gly-31, Gln-149 to
Ser-155.
[0514] The hematopoetic tissue distribution and homology to a cell
surface molecule indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the detection and/or
treatment of immune or hematopoietic disorders including arthritis,
asthma and immunodeficiency diseases. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0515] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:79 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 2441 of SEQ ID NO:79, b is an integer
of 15 -to 2455, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:79, and where b is greater
than or equal to a+14.
[0516] Features of Protein Encoded by Gene No: 70
[0517] The gene encoding the disclosed cDNA is thought to reside on
chromosome 4. Accordingly, polynucleotides related to this
invention are useful as a marker in linkage analysis for chromosome
4. When tested against a Jurkat T-cell line, supernatants removed
from cells containing this gene activated the GAS (gamma activation
site) pathway. Thus, it is likely that this gene activates T-cells
through the Jaks-STAT signal transduction pathway. The GAS is a
promoter element found upstream in many genes which are involved in
the Jaks-STAT pathway. The Jaks-STAT pathway is a complex, signal
transduction pathway involved in the differentiation and
proliferation of cells. Therefore, activation of the Jaks-STATs
pathway, reflected by the binding of the GAS element, can be used
to indicate proteins involved in the proliferation and
differentiation of cells.
[0518] This gene is expressed primarily in brain.
[0519] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, neurodegenerative conditions and behavioural disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the brain and central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., brain and other tissues of the
nervous system, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0520] Preferred epitopes include those comprising a sequence shown
in-SEQ ID NO. 219 as residues: Asp-57 to Gly-64.
[0521] The tissue distribution of this gene primarily in brain
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the treatment and/or detection of
neurodegenerative disease states and behavioural disorders such as
Alzheimers Disease, Parkinsons Disease, Huntingtons Disease,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder and panic disorder. In addition, the gene or gene product
may also play a role in the treatment and/or detection of
developmental disorders associated with the developing embryo, or
sexually-linked disorders. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0522] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:80 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 907 of SEQ ID NO:80, b is an integer
of 15 to 921, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:80, and where b is greater
than or equal to a+14.
[0523] Features of Protein Encoded by Gene No: 71
[0524] This gene is expressed primarily in hematopoietic progenitor
cells (CD34+ cells).
[0525] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, autoimmune and immunodeficiency disease states.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the hematopoietic system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types and cell types (e.g., hematopoietic cells, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0526] The tissue distribution of this gene predominantly in
hematopoietic progenitor cell types indicates that the gene could
be important for the treatment or detection of immune or
hematopoietic disorders including arthritis, asthma,
immunodeficiency diseases, leukemia, hypersensitivity and
transplant rejection. Additionally, expression of this gene product
in CD34+ cells indicates a role in the regulation of the
proliferation; survival; differentiation; and/or activation of
potentially all hematopoietic cell lineages, including blood stem
cells. This gene product may be involved in the regulation of
cytokine production, antigen presentation, or other processes that
may also suggest a usefulness in the treatment of cancer (e.g., by
boosting immune responses).
[0527] Since the gene is expressed in cells of lymphoid origin, the
gene or protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues Therefore it may be also used
as an agent for immunological disorders including arthritis,
asthma, immune deficiency diseases such as AIDS, leukemia,
rheumatoid arthritis, inflammatory bowel disease, sepsis, acne, and
psoriasis. In addition, this gene product may have commercial
utility in the expansion of stem cells and committed progenitors of
various blood lineages, and in the differentiation and/or
proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0528] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:81 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 664 of SEQ ID NO:81, b is an integer
of 15 to 678, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:81, and where b is greater
than or equal to a+14.
[0529] Features of Protein Encoded by Gene No: 72
[0530] This gene is expressed primarily in hematopoietic progenitor
cells (CD34+ cells).
[0531] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, auto-immune and immunodeficiency disease states.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the hematopoietic system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types and cell types (e.g., hematopoietic cells, and
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0532] The tissue distribution of this gene predominantly in
hematopoietic progenitor cell types indicates that the gene is
important for the treatment or detection of immune or hematopoietic
disorders including arthritis, asthma, immunodeficiency diseases,
leukemia, and transplant rejection. Expression of this gene product
in CD34+ cells indicates a role in the regulation of the
proliferation; survival; differentiation; and/or activation of
potentially all hematopoietic cell lineages, including blood stem
cells. This gene product may be involved in the regulation of
cytokine production, antigen presentation, or other processes that
may also suggest a usefulness in the treatment of cancer (e.g. by
boosting immune responses).
[0533] Since the gene is expressed in cells of lymphoid origin, the
gene or protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues. Therefore it may be also used
as an agent for immunological disorders including arthritis,
asthma, immune deficiency diseases such as AIDS, leukemia,
rheumatoid arthritis, inflammatory bowel disease, sepsis, acne, and
psoriasis. In addition, this gene product may have commercial
utility in the expansion of stem cells and committed progenitors of
various blood lineages, and in the differentiation and/or
proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0534] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 82 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 843 of SEQ ID NO:82, b is an integer
of 15 to 857, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:82, and where b is greater
than or equal to a+14.
[0535] Features of Protein Encoded by Gene No: 73
[0536] The translation product of this gene shares sequence
homology with rat synaptogyrin which is thought to be important in
membrane trafficking (see e.g., Stenius et al., J. Cell Biol. 131
(6 Pt 2), 1801-1809 (1995)). In specific embdodiments, polypeptides
of the invention comprise the following amino acid sequences:
26 QPYQVLPSRQVFALI (SEQ ID NO:508), VFSCIYGEGYSNAHESKQMYCVFN (SEQ
ID NO:509), RNEDACRYGSAIGVLAFL (SEQ ID NO:510), LVVDAYFPQISNATDRK
(SEQ ID NO:511), and/or SALWTFLWFVGFCFLTNQWAVTNPK (SEQ ID
NO:512).
[0537] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0538] This gene is expressed primarily in breast and ovary, and to
a lesser extent in most hematopoietic tissue types.
[0539] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, female infertility and female reproductive
abnormalities. Similarly, polypeptides and antibodies directed to
these polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the reproductive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., mammary tissue, and ovary and other
reproductive tissue, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0540] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 222 as residues: Pro-9 to Trp-18, Thr-20 to
Ala-27.
[0541] The tissue distribution in ovary and breast and homology to
a protein involved in membrane trafficking indicates that this
protein may play a role in the detection/treatment of female
fertility disorders, endocrine disorders, ovarian failure,
amenorrhea, ovarian cancer, and also potentially in both
non-neoplastic breast diseases such as congenital abnormalities and
neoplasia in the breast. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0542] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:83 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1963 of SEQ ID NO:83, b is an integer
of 15 to 1977, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:83, and where b is greater
than or equal to a+14.
[0543] Features of Protein Encoded by Gene No: 74
[0544] The gene encoding the disclosed cDNA is thought to reside on
chromosome 12. Accordingly, polynucleotides related to this
invention are useful as a marker in linkage analysis for chromosome
12. In specific embodiments, polypeptides of the invention comprise
the following amino acid sequence:
27 LNIDSFDYGKFESLLAKQHYKFSFLLPLAAGTERCKWWLKIEEASSDQCGCWFLVKCVPKPP-
SPCRQPPTQVSKIGHAPFFL (SEQ ID NO:513).
[0545] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0546] This gene is expressed primarily in brain, and to a lesser
extent in placenta and spleen.
[0547] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, behavioural disorders and neurodegenerative disease
states. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly. of
the brain and central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., brain and other tissue of the
nervous system, spleen and other cells and tissue of the immune
system, placenta, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0548] The tissue distribution in brain indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, schizophrenia, mania,
dementia, paranoia, obsessive compulsive disorder and panic
disorder. In addition, the gene or gene product may also play a
role in the treatment and/or detection of developmental disorders
associated with the developing embryo, or sexually-linked
disorders. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0549] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 84 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1135 of SEQ ID NO:84, b is an integer
of 15 to 1149, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:84, and where b is greater
than or equal to a+14.
[0550] Features of Protein Encoded by Gene No: 75
[0551] When tested against a K562 cell line, supernatants removed
from cells containing this gene activated the ISRE
(interferon-sensitive responsive element) pathway. Thus, it is
likely that this gene activates leukemia cells, or more generally,
in immune or hematopoietic cells, or other cells or cell-types,
through the Jaks-STAT signal transduction pathway. The ISRE is a
promoter element found upstream in many genes which are involved in
the Jaks-STAT pathway. The Jaks-STAT pathway is a complex, signal
transduction pathway involved in the differentiation and
proliferation of cells. Therefore, activation of the Jaks-STATs
pathway, reflected by the binding of the GAS element, can be used
to indicate proteins involved in the proliferation and
differentiation of cells.
[0552] This gene is expressed primarily in bone marrow and
spleen.
[0553] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, autoimmune diseases, transplant rejection and
immundeficiency disease states. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the hematopoietic and immune
systems, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., immune, cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0554] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 224 as residues: Pro-22 to His-33, Ser-42 to
Trp-48.
[0555] The tissue distribution of this gene predominantly in
hematopoietic cell types indicates that the gene is important for
the treatment or detection of immune or hematopoietic disorders
including arthritis, asthma, immunodeficiency diseases, leukemia,
and transplant rejection. Furthermore, the polypeptides or
polynucleotides are also useful to enhance or protect the
proliferation, differentiation, and functional activation of
hematopoietic progenitor cells (e.g., bone marrow cells), useful in
treating cancer patients undergoing chemotherapy or patients
undergoing bone marrow transplantation. The polypeptides or
polynucleotides are also useful to increase the proliferation of
peripheral blood leukocytes, which can be used in the combat of a
range of hematopoietic disorders, including immunodeficiency
diseases, leukemia, and septicemia.
[0556] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 85 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence. would be cumbersome.
[0557] Accordingly, preferably excluded from the present invention
are one or more polynucleotides comprising a nucleotide sequence
described by the general formula of a-b, where a is any integer
between 1 to 753 of SEQ ID NO:85, b is an integer of 15 to 767,
where both a and b correspond to the positions of nucleotide
residues shown in SEQ ID NO:85, and where b is greater than or
equal to a+14.
[0558] Features of Protein Encoded by Gene No: 76
[0559] In specific embodiments, polypeptides of the invention
comprise the sequence:
28 SLQYRIRIPGRPT (SEQ ID NO:514), DLVTYTSSLQYRIRIPGRPTRP (SEQ ID
NO:515), VKTAECYSIPLGSCPVNIQRVR (SEQ ID NO:517), and/or
LGNKKYINIRCLEMQVTLKILCEIEKKERRGTHCLV (SEQ ID NO:516).
[0560] Polynucleotides encoding these polypeptides are also
encompassed by the invention. Contact of cells with supernatant
expressing the product of this gene increases the permeability of
U937 monocyte cells to calcium. Thus, it is likely that the product
of this gene is involved in a signal transduction pathway that is
initiated when the product of this gene binds a receptor on the
surface of the monocyte cell. Thus, polynucleotides and
polypeptides have uses which include, but are not limited to,
activating monocyte cells.
[0561] This gene is expressed in primary dendritic cells.
[0562] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, auto-immune disorders such as asthma and arthritis, in
transplant rejection, leukemia and immunodeficiency disease states.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune system, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types and cell types (e.g., primary dendritic cells and other
cells and tissue of the immune system, and cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0563] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 225 as residues: Gly-2 to Glu-7, Arg-27 to
Gly-34.
[0564] The tissue distribution of this gene predominantly in
hematopoietic cell types indicates that the gene is important for
the treatment or detection of immune or hematopoietic disorders
including arthritis, asthma, immunodeficiency diseases, leukemia,
hypersensitivity and graft rejection. Expression of this gene
product in primary dendritic cells also strongly indicates a role
for this protein in immune function and immune surveillance.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0565] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:86 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 714 of SEQ ID NO:86, b is an integer
of 15 to 728, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:86, and where b is greater
than or equal to a+14.
[0566] Features of Protein Encoded by Gene No: 77
[0567] This gene is expressed primarily in 12 week old early stage
human.
[0568] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, developmental abnormalities. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the developmental system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
developing and differentiating tissue, and cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0569] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 226 as residues: Thr-14 to Thr-19.
[0570] The expression of this gene primarily in the embryo
indicates a key role in embryonic development, and could be used in
the treatment and or detection of developmental disorders.
Expression within embryonic tissue and other cellular sources
marked by proliferating cells indicates that this protein may play
a role in the regulation of cellular division, and may show utility
it the diagnosis and treatment of cancer and other proliferative
disorders. Similarly, embryonic development also involves decisions
involving cell differentiation and/or apoptosis in pattern
formation. Thus this protein may also be involved in apoptosis or
tissue differentiation and could again be useful in cancer therapy.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0571] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:87 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 721 of SEQ ID NO:87, b is an integer
of 15 to 735, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:87, and where b is greater
than or equal to a+14.
[0572] Features of Protein Encoded by Gene No: 78
[0573] This gene is expressed primarily in T-cells, and to a lesser
extent in cord blood and osteosarcoma.
[0574] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, auto-immune diseases, immunodeficiency diseases and
host-graft rejection. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., cells and tissues
of the immune system, bone, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0575] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 227 as residues: Pro-36 to Ala-41.
[0576] The expression of this gene in T-cells indicates a potential
role in the treament/detection of immune disorders such as
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia and transplant rejection. Expression of this gene product
in T cells also strongly indicates a role for this protein in
immune function and immune surveillance. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0577] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:88 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 875 of SEQ ID NO:88, b is an integer
of 15 to 889, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:88, and where b is greater
than or equal to a+14.
[0578] Features of Protein Encoded by Gene No: 79
[0579] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
29 LFYLLTCSCAPGHLAFVCSQCLPFDMGKELWPKSPSSCTSTSVAQGWGGRGRPSPYICVV
(SEQ ID NO:518), IQGSRLPPLPAPLHPLPLIYLLLGSPAQSWLLVPSWGHP-
STLTLTMAAEHQAWPSGFHGDH (SEQ ID NO:519).
[0580] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0581] This gene is expressed primarily in placenta and 9 week old
embryo, and to a lesser extent in fetal spleen.
[0582] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, developmental disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the developmental system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
developing and differentiating tissues, and spleen and other cells
and tissue of the immune system, and cancerous and wounded tissues)
or bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0583] The expression of this gene primarily in the embryo
indicates a key role in embryonic development, and could be used in
the treatment and or detection of developmental disorders. The
tissue distribution in placenta also indicates that polynucleotides
and polypeptides corresponding to this gene are useful for the
diagnosis and/or treatment of disorders of the placenta. Specific
expression within the placenta indicates that this gene product may
play a role in the proper establishment and maintenance of
placental function. Alternately, this gene product may be produced
by the placenta and then transported to the embryo, where it may
play a crucial role in the development and/or survival of the
developing embryo or fetus. Expression within embryonic tissue and
other cellular sources marked by proliferating cells indicates that
this protein may play a role in the regulation of cellular
division. Similarly, embryonic development also involves decisions
involving cell differentiation and/or apoptosis in pattern
formation. Thus this protein may also be involved in apoptosis or
tissue differentiation and could again be useful in cancer therapy.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0584] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 89 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 555 of SEQ ID NO:89, b is an integer
of 15 to 569, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:89, and where b is greater
than or equal to a+14.
[0585] Features of Protein Encoded by Gene No: 80
[0586] This gene is expressed primarily in early stage brain.
[0587] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, developmental and neurodegenerative diseases of the
brain and nervous system. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the brain, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., brain and other
tissue of the nervous system, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0588] The tissue distribution in brain indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment and detection of developmental and
neurodegenerative diseases, as well as behavioral or nervous system
disorders.
[0589] Examples of such conditions would include: depression,
schizophrenia, mania, dementia, paranoia, addictive behavior and
sleep disorders. In addition a brain-specific gene product may be
useful in the diagnosis of specific brain tumors. Protein, as well
as, antibodies directed against the protein may show utility as a
tumor marker and/or immunotherapy targets for the above listed
tissues.
[0590] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:90 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 320 of SEQ ID NO:90, b is an integer
of 15 to 334, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:90, and where b is greater
than or equal to a+14.
[0591] Features of Protein Encoded by Gene No: 81
[0592] This gene is expressed primarily in synovial tissue.
[0593] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, arthritis, tendonitis and chrondomalacia. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
synovium, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., synovial tissue, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0594] The tissue distribution in synovial tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment and diagnosis of connective tissue
disorders such as arthritis, tendonitis, chrondomalacia,
inflammation and trauma. In addition, the expression of this gene
product in synovium indicates a role in the detection and treatment
of disorders and conditions affecting the skeletal system, in
particular osteoporosis as well as disorders afflicting connective
tissues (e.g. arthritis, trauma, tendonitis, chrondomalacia and
inflammation), such as in the diagnosis or treatment of various
autoimmune disorders such as rheumatoid arthritis, lupus,
scleroderma, and dermatomyositis as well as dwarfism, spinal
deformation, and specific joint abnormalities as well as
chondrodysplasias (ie. spondyloepiphyseal dysplasia congenita,
familial osteoarthritis, Atelosteogenesis type II, metaphyseal
chondrodysplasia type Schmid). Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0595] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:91 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 781 of SEQ ID NO:91, b is an integer
of 15 to 795, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:91, and where b is greater
than or equal to a+14.
[0596] Features of Protein Encoded by Gene No: 82
[0597] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence: VDPPGCRNSARGCTRLLRGSSKI
(SEQ ID NO: 520). Polynucleotides encoding these polypeptides are
also encompassed by the invention.
[0598] This gene is expressed primarily in the frontal cortex of
the brain.
[0599] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, developmental and neurodegenerative diseases of the
brain. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the central nervous system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., brain and other tissue of the
nervous system, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0600] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 231 as residues: Ser-4 to Tyr-13.
[0601] The tissue distribution in brain indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection and treatment of developmental and
neurodegenerative diseases of the brain and nervous system,
including malignancies as well as behavioral disorders. Examples of
such conditions might include: depression, schizophrenia,
Alzheimer's disease, Parkinson's disease, Huntington's disease,
mania, dementia, paranoia, addictive behavior and sleep disorders.
Furthermore, elevated expression of this gene product within the
frontal cortex of the brain indicates that it may be involved in
neuronal survival; synapse formation; conductance; neural
differentiation, etc. Such involvement may impact many processes,
such as learning and cognition. It may also be useful in the
treatment of such neurodegenerative disorders as schizophrenia;
ALS; or Alzheimer's.
[0602] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:92 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one-or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 563 of SEQ ID NO:92, b is an integer
of 15 to 577, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:92, and where b is greater
than or equal to a+14.
[0603] Features of Protein Encoded by Gene No: 83
[0604] The translation product of this gene shares sequence
homology with the L6 cell surface antigen, which is highly
expressed in lung, breast, colon, and ovarian carcinomas. See e.g.,
Marken et al., Proc Natl Acad Sci U S A 1992 April
15;89(8):3503-3507. In specific embodiments, polypeptides of the
invention comprise the sequence: ITLCLVCIVANA (SEQ ID NO: 521).
Polynucleotides encoding these polypeptides are also encompassed by
the invention. This gene was recently cloned and sequenced by
another group, which identified the gene as a putative tetraspan
transmembrane (TM4) protein L6H from humans. The transmembrane 4
superfamily (TM4SF) or tetraspan superfamily has at least 16
members (including CD9, CD20, CD37, CD53, CD63, CD81, CD82, A15,
CO-029, Sm23, RDS, Uro B, Uro A, SAS, Rom-1, PETA3, and YKK8), is
the second biggest subfamily among CD antigen superfamilies, and
are activation antigens of T-cells. All TM4SF members contain four
putative transmembrane domains, two extracellular loops, and two
short cytoplasmic tails. They are variously expressed on immature,
early, mature, activated lymphocytes, monocytes, macrophages,
granulocytes, platelets, eosinophils, basophils, certain leukemic
and lymphoma cells, and a variety of other cells and tissues.
[0605] This gene is expressed primarily in fetal liver/spleen
tissues.
[0606] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, cancers of the liver, immune system disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the hepatic and immune system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., lung and pulmonary tissue,
colon and other gastrointestinal tissue, mammary tissue, ovarian
tissue and other tissue of the reproductive system, hepatic tissue,
immune system tissues, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0607] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 232 as residues: Asn-32 to Asn-41, Thr-140 to
Ala-147, Asp-188 to His-197.
[0608] The murine monoclonal antibody (mAb) L6 recognizes an
integral membrane glycoprotein that is highly expressed in lung,
breast, colon, and ovarian carcinomas and is referred to as the L6
antigen. This antigen is an attractive target for therapeutic
intervention due to its high level expression on malignant cells.
The tissue distribution and homology to L6 antigen indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for detection and treatment of neoplastic tissues
particularly of the liver. The translation product of this gene is
a member of the tetraspan transmembrane superfamily, and therefore,
antigenic regions of members of this family could be valuable
immunogens or targets to implement active and passive immunotherapy
in patients with cancer. Moreover, the protein product of this
clone is useful for the treatment and diagnosis of hematopoietic
related disorders such as anemia, pancytopenia, leukopenia,
thrombocytopenia or leukemia since stromal cells are important in
the production of cells of hematopoietic lineages. The uses include
bone marrow cell ex-vivo culture, bone marrow transplantation, bone
marrow reconstitution, radiotherapy or chemotherapy of neoplasia.
The gene product may also be involved in lymphopoiesis, therefore,
it can be used in immune disorders such as infection, inflammation,
allergy, immunodeficiency etc. In addition, this gene product may
have commercial utility in the expansion of stem cells and
committed progenitors of various blood lineages, and in the
differentiation and/or proliferation of various cell types.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0609] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:93 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 954 of SEQ ID NO:93, b is an integer
of 15 to 968, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:93, and where b is greater
than or equal to a+14.
[0610] Features of Protein Encoded by Gene No: 84
[0611] This gene is expressed primarily in glioblastoma.
[0612] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, glioblastoma. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the central nervous system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
tissue of the nervous system, and cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0613] The tissue distribution in glioblastoma indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection and treatment of malignancies, as well as
developmental and neurodegenerative diseases of the brain and
nervous system. Protein, as well as, antibodies directed against
the protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0614] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:94 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 539 of SEQ ID NO:94, b is an integer
of 15 to 553, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:94, and where b is greater
than or equal to a+14.
[0615] Features of Protein Encoded by Gene No: 85
[0616] The translation product of this gene shares sequence
homology with Tbx, which is thought to be important in
developmental regulation (see e.g., Knezevic et al., Development
124, 411-419 (1997); and accession U80951). In specific
embodiments, polypeptides of the invention comprise the
sequence:
30 VTAYQNQQITRLKIDRNPFAKGFR (SEQ ID NO:522), GTATVTAYQNQQITRL (SEQ
ID NO:523), KIDRNPFAKGFRDSGRNRMGLEA- L (SEQ ID NO:524),
STLLQVLGMAFLPLTLTFCLA (SEQ ID NO:525), and/or
VESYAFWRPSLRTLTFEDIPGIPKQGNASS (SEQ ID NO:526).
[0617] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0618] This gene is expressed primarily in synovial sarcoma and to
a lesser extent in osteoclastoma, osteoblastoma, and
hemangiopericytoma.
[0619] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, osteosarcoma, osteoclastoma, and chondrosarcoma, and
diseases of the skeletal system, such as osteoporosis. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
skeletal system, expression of this gene at significantly higher or
lower levels may be routinely detected in certain tissues or cell
types and cell types (e.g., bone cells and tissue, synovial cells
and tissue, cartilage, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0620] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 234 as residues: Ala-45 to Asp-50, Arg-57 to
Pro-63.
[0621] The tissue distribution in skeletal tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment and diagnosis of osteoperosis, fracture,
osteosarcoma, osteoclastoma, chondrosarcoma, ossification and
osteonecrosis, arthritis, tendonitis, chrondomalacia, and
inflammation. Elevated levels of expression of this gene product in
osteoclastoma and osteoblastoma indicates that it may play a role
in the survival, proliferation, and/or growth of these cells.
Therefore, it may be useful in influencing bone mass in such
conditions as osteoporosis. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0622] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:95 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 954 of SEQ ID NO:95, b is an integer
of 15 to 968, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:95, and where b is greater
than or equal to a+14.
[0623] Features of Protein Encoded by Gene No: 86
[0624] The gene encoding the disclosed cDNA is thought to reside on
chromosome 19. Accordingly, polynucleotides related to this
invention are useful as a marker in linkage analysis for chromosome
19. The translation product of this gene is a transmembrane protein
that forms disulfide-bonded homodimers and contains a motif in its
cytoplasmic domain (located at the carboxy terminus of the protein
relative to the transmembrane domain) that functions as an adaptor
for associating protein complexes involved in triggering cellular
activation. The transmembrane domain is predicted to consist of the
amino acid sequence: VLAGIVMGDLVLTVLIALAVYFLG.(SEQ ID NO: 528). In
specific embodiments, polypeptides of the invention comprise the
following amino acid sequences:
31 QAQSDCSCSTVSPG (SEQ ID NO:527), VLAGIVMGDLVLTVLIALAVYFLG (SEQ ID
NO:528), VPRGRGAAEATRKQRITETESPYQELQGQRSDVYSDL (SEQ ID NO:529),
and/or ETESPYQELQGQRSDVYSDLNT (SEQ ID NO:530).
[0625] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0626] This gene is expressed primarily in macrophage, and to a
lesser extent in primary dendritic cells and neutrophils.
[0627] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immunologically mediated disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
and cell types (e.g., blood cells, and cells and tissue of the
immune system, and cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0628] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 235 as residues: Ala-28 to Ser-33, Ala-76 to
Lys-111.
[0629] The tissue distribution in immune tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of immune
disorders including: leukemias, lymphomas, auto-immunities,
immunodeficiencies (e.g., AIDS), immuno-supressive conditions
(transplantation) and hematopoietic disorders. Furthermore,
expression of this gene product in macrophage and primary dendritic
cells also strongly indicates a role for this protein in immune
function and immune surveillance. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0630] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:96 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 683 of SEQ ID NO:96, b is an integer
of 15 to 697, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:96, and where b is greater
than or equal to a+14.
[0631] Features of Protein Encoded by Gene No: 87
[0632] This gene is expressed primarily in prostate cancer.
[0633] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, prostate cancer. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the prostate, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., prostate,
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0634] The tissue distribution in prostate cancerous tissue
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the detection and treatment of prostate
cancer and other prostate disorders, as well as cancers in other
tissues where expression has been indicated. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0635] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:97 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 852 of SEQ ID NO:97, b is an integer
of 15 to 866, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:97, and where b is greater
than or equal to a+14.
[0636] Features of Protein Encoded by Gene No: 88
[0637] The translation product of this gene shares sequence
homology with retinal epithelial membrane protein (REMP), which is
thought to be important in development and maintenance of normal
retinal function (See e.g., Philp et al., Exp. Cell Res. 219 (1),
64-73 (1995); and Genbank Accesion No.U15685). The translation
product of this gene also shares homology with monocarboxylate
transporter protein (Genbank Accesion no.U87627). Another group
recently cloned and sequenced this gene, describing it as a
monocarboxylate transporter protein (Genbank Accession No.
gi.vertline.2463634). In quantitative terms, lactic acid is one of
the most important metabolites in the body, substantial amounts
being used and/or produced by almost all mammalian cells. As such
it must be rapidly transported into and out of cells. Lactic acid
transport across the plasma membrane is catalysed by proton-linked
monocarboxylate transporters (MCTs), which are also responsible for
the transport of pyruvate and the ketone bodies acetoacetate,
-hydroxybutyrate and acetate. In specific embodiments, polypeptides
of the invention comprise the following amino acid sequence:
32 FLCALSPLGQLLQDRYGWRGGFLILGGL, (SEQ ID NO:531)
LLNCCVCAALMRPLVVTAQPGXGPPRP, (SEQ ID NO:532) and/or
SRRLXDLSVFRDRGFVLYAVAASVM (SEQ ID NO:533).
[0638] Polynucleotides encoding these polypeptides are also
encompassed by the invention. The gene encoding the disclosed cDNA
is thought to reside on chromosome 17. Accordingly, polynucleotides
related to this invention are useful as a marker in linkage
analysis for chromosome 17.
[0639] This gene is expressed primarily in neutrophils, and to a
lesser extent in a variety of other tissues and cell types,
including retina.
[0640] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, eye, and metabolic and cellular transport disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly. of
the eye and immune system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types and cell types (e.g., retinal cells, neutrophils and
other blood cells, and cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0641] The tissue distribution in retinal tissue and the homology
to REMP indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the diagnosis, treatment,
and/or prevention of eye disorders, including neoplasms, visual
impairments and blindness. Alternatively, the homology to
monocarboxylate transporter protein indicates that the translation
product of this gene is useful for the diagnosis and/or treatment
of disorders involving the cellular transport of lactic acid into
and out of the cell.
[0642] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:98 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1354 of SEQ ID NO:98, b is an integer
of 15 to 1368, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:98, and where b is greater
than or equal to a+14.
[0643] Features of Protein Encoded by Gene No: 89
[0644] The translation product of this gene shares sequence
homology with human squamous cell E48 antigen which is thought to
be important in self-recognition and immune function. In specific
embodiments, polypeptides of the invention comprise the following
amino acid sequences:
33 MMATPSTRPPPPAASTTSATAPALPPRPPWPWPPSSWPPSGVSSKAPEADPLKNKAL (SEQ
ID NO:534); LLLTSPLPRCPPACSHDAPAHPDPGGPHGLTSGPGLGLPRVCLQ-
RRQLLQPHALPGYGCLLHDHAHLLHPHQDEGQ (SEQ ID NO:535); and/or
WLLQARVHHLLLPVRPLQRHRPCHPGHPGPGPHPPGHPLGSPLKPPRQTHSRTKLS (SEQ ID
NO:536).
[0645] Polynucleotides encoding these polypeptides are also
encompassed by the invention. When tested against K562 leukemia
cell lines, supernatants removed from cells containing this gene
activated the ISRE assay. Thus, it is likely that this gene
activates leukemia cells through the Jak-STAT signal transduction
pathway. The interferon-sensitive response element is a promoter
element found upstream of many genes which are involved in the
Jak-STAT pathway. The Jak-STAT pathway is a large, signal
transduction pathway involved in the differentiation and
proliferation of cells. Therefore, activation of the Jak-STAT
pathway, reflected by the binding of the ISRE element, can be used
to indicate proteins involved in the proliferation and
differentiation of cells.
[0646] This gene is expressed primarily in adult brain, and to a
lesser extent in fetal lung.
[0647] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, autoimmune disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0648] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 238 as residues: Tyr-28 to Phe-34, Thr-54 to Val-60,
Tyr-73 to Thr-82.
[0649] The tissue distribution and homology to human squamous cell
E48 antigen indicates that polynucleotides and polypeptides
corresponding to this gene are useful for study, diagnosis and
treatment of autoimmune diseases and disorders, such as lupus,
transplant rejection, allergic reactions, and arthritis. Protein,
as well as, antibodies directed against the protein may show
utility as a tissue-specific marker and/or immunotherapy target for
the above listed tissues.
[0650] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:99 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 599 of SEQ ID NO:99, b is an integer
of 15 to 613, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:99, and where b is greater
than or equal to a+14.
[0651] Features of Protein Encoded by Gene No: 90
[0652] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
34 QEFQTGLGNMVKPCLYEKYRNISWLWWHTPVVPATWEAEVGGSLEPGRLRLQ (SEQ ID
NO:537), and/or ILGGESILIILSWVFSYIFFRIALEITIYILNVSPFCLGR-
WLMPVIPALWEAEVGGLPELRSSRPA (SEQ ID NO:538).
[0653] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0654] This gene is expressed primarily in human adult lymph node
tissue.
[0655] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune disorders and lymphomas. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune and metabolic
systems, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., immune, metabolic, cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0656] The tissue distribution in lymph nodes indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study, diagnosis and treatment of immune and lymph
diseases and disorders such as lymphomas. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer (e.g. by boosting immune responses).
Since the gene is expressed in cells of lymphoid origin, the
natural gene product may be involved in immune functions. Therefore
it may be also used as an agent for immunological disorders
including arthritis, asthma, immunodeficiency diseases such as
AIDS, leukemia, rheumatoid arthritis, granulomatous disease,
inflammatory bowel disease, sepsis, acne, neutropenia,
neutrophilia, psoriasis, hypersensitivities, such as T-cell
mediated cytotoxicity; immune reactions to transplanted organs and
tissues, such as host-versus-graft and graft-versus-host diseases,
or autoimmunity disorders, such as autoimmune infertility, lense
tissue injury, demyelination, systemic lupus erythematosis, drug
induced hemolytic anemia, rheumatoid arthritis, Sjogren's disease,
scleroderma and tissues. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0657] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:100 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 671 of SEQ ID NO: 100, b is an
integer of 15 to 685, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO:100, and where
b is greater than or equal to a+14.
[0658] Features of Protein Encoded by Gene No: 91
[0659] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
35 MPKQLAQLLYRLPRG, (SEQ ID NO:539) LFQAISVSGSHR, (SEQ ID NO:540)
QGSRTWNTLTEGNAEAACTVALQTSKRLILASRW TLSFM, (SEQ ID NO:541)
NSHCVPIKALFFLSVVSYIFIMPHHIFFTVKILKSCFQVGQLM- KL and/or
RPTRPITFSSNISEWVPSTGFQDLEHFNRRKCRSSLHSCFT- DFQEA (SEQ ID NO:542)
DSGFKMEPWSWFFFFFFFFPQRTCGCALCVLFLFSIWGPHGKELL- NSFLYELPL
CSYKGPFLS.
[0660] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0661] This gene is expressed primarily in placenta and
synovium.
[0662] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases of the synovium and placenta. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
placenta and synovium, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., placenta, synovium, cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0663] The tissue distribution in placenta and synovium indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for the study, diagnosis and treatment of growth and
developmental disorders and arthritic and inflammatory conditions.
Expression within embryonic tissue and other cellular sources
marked by proliferating cells indicates that this protein may play
a role in the regulation of cellular division, and may show utility
in the diagnosis and treatment of cancer and other proliferative
disorders. Similarly, embryonic development also involves decisions
involving cell differentiation and/or apoptosis in pattern
formation. Thus this protein may also be involved in apoptosis or
tissue differentiation and could again be useful in cancer therapy.
Specific expression within the placenta indicates that this gene
product may play a role in the proper establishment and maintenance
of placental function. Alternately, this gene product may be
produced by the placenta and then transported to the embryo, where
it may play a crucial role in the development and/or survival of
the developing embryo or fetus. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0664] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:101 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 632 of SEQ ID NO:101, b is an integer
of 15 to 646, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO: 101, and where b is greater
than or equal to a+14.
[0665] Features of Protein Encoded by Gene No: 92
[0666] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
36 VDPRVRLPLFWWQPSCAVYLFPRVYNNMCTRVLGTLPHCWDLATLLQPSSRI, (SEQ ID
NO:543) WGNVSEAPGM VPYHIAGTLPHCCSLPVGYGGMSVRLQ, (SEQ ID NO:544)
GCRYVGNVGPQGNMQSGRSWALKMVLLCNSCLGLGVGSVGPSMSSLFGAV- L SETPGSSVY
MLDPRATCNLVGVGLSKWCCCVTAAWVLG (SEQ ID NO:545).
[0667] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0668] This gene is expressed primarily in chronic lymphocytic
leukemia.
[0669] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases of immune system including cancer. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., immune, cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0670] The tissue distribution in chronic lymphocytic leukemia
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the study, diagnosis and treatment of
disorders of the immune system including cancers. Therefore it may
be also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia, rheumatoid arthritis, inflammatory bowel disease, sepsis,
acne, and psoriasis. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0671] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:102 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 812 of SEQ ID NO: 102, b is an
integer of 15 to 826, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO:102, and where
b is greater than or equal to a+14.
[0672] Features of Protein Encoded by Gene No: 93
[0673] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
37
HGDWIYVHIVEQLNQANNKSVTSHTYFVVKTCKIHSLSNFQASNTLLXTVVTMLYNRSLELILP- V
(SEQ ID NO:546) LYNRSLELILPV TYSSCLTKILYSLINIYPIPHCSPAXITTILL, (SEQ
ID NO:547) SMNLTFFFFRFHICEIAQYLSFCAWLISLNIKSL and/or
MNLTFFFFRFHICEIAQYLSFCAWLISLNIKSL (SEQ ID NO:548).
[0674] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0675] This gene is expressed primarily in brain
medulloblastoma.
[0676] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of cancer and disorders of the CNS. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
central nervous system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., brain, cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0677] The tissue distribution in brain indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis, treatment, and/or prevention of cancers
and other disorders and diseases of the CNS. The tissue
distribution further indicates that polynucleotides and
polypeptides corresponding to this gene are useful for the
detection/treatment of neurodegenerative disease states and
behavioural disorders such as Alzheimers Disease, Parkinsons
Disease, Huntingtons Disease, Tourette Syndrome, schizophrenia,
mania, dementia, paranoia, obsessive compulsive disorder, panic
disorder, learning disabilities, ALS, psychoses, autism, and
altered bahaviors, including disorders in feeding, sleep patterns,
balance, and perception. In addition, the gene or gene product may
also play a role in the treatment and/or detection of developmental
disorders associated with the developing embryo, or sexually-linked
disorders. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0678] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 103 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 572 of SEQ ID NO: 103, b is an
integer of 15 to 586, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO: 103, and where
b is greater than or equal to a+14.
[0679] Features of Protein Encoded by Gene No: 94
[0680] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
38 LVCYCSTKKEKKLHEIAIQQGQNWRWLLFYKEISVPGFQSVWCSYKCLCVVW, (SEQ ID
NO:549) KAGEGG RRSCSGPPLVNTAGKILSSSPAKLACKRTDFHIP- , (SEQ ID
NO:550) SI RASILGIDNERGCHFRHFNPLKEYKRKKKE- NKSFRIV, (SEQ ID NO:551)
SKNKTRGGDWCVTVLRKRRKSFMKSPFSKDRTG- DGF, (SEQ ID NO:552)
SFTKKSLSQAFSLFGVHTSVCVLCGRRGKAGEGGPVQGPLW and/or
MKSPFSKDRTGDGFSFTKKSLSQAFSLFGVHTSVCVLCGR (SEQ ID NO:553)
RGKAGEGGPVQGPLW.
[0681] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0682] This gene is expressed primarily in meningima and
neutrophils and to a lesser extent in anergic T cells and CD34
depleted buffy coat.
[0683] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, inflammatory, immune and hemopoietic disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the hemopoietic, immune and inflammatory systems, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0684] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 243 as residues: Glu-45 to Asn-50.
[0685] The tissue distribution in immune tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study, diagnosis and treatment of various disorders
and diseases of the immune, inflammatory, and hemopoietic systems.
Furthermore, the tissue distribution indicates that polynucleotides
and polypeptides corresponding to this gene are useful for the
diagnosis and/or treatment of hematopoietic disorders. This gene
product is primarily expressed in hematopoietic cells and tissues,
suggesting that it plays a role in the survival, proliferation,
and/or differentiation of hematopoieitic lineages. Expression of
this gene product in T cells and neutrophils also strongly
indicates a role for this protein in immune function and immune
surveillance. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0686] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:104 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 614 of SEQ ID NO:104, b is an integer
of 15 to 628, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:104, and where b is greater
than or equal to a+14.
[0687] Features of Protein Encoded by Gene No: 95
[0688] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
39 MGESECYRRLSGASCTWTVHVDFA (SEQ ID NO:554);
MHCGTRVWKTMKHDYFLLACLSMTSTGGILCTL 9SEQ ID NO:555); STLSLI (SEQ ID
NO:556); and/or PTSSSLSFWPWCTAIIGSIFTYCVCVCVCFVVMNRTCYLPNS-
IIYHNSKLATIIDK SMTLS MWILPKVSLICIVELGYGKP (SEQ ID NO:557).
[0689] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0690] This gene is expressed primarily in human meningima.
[0691] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, meningitis and other inflammatory conditions.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the cerebrospinal membranes, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., cancerous and wounded tissues)
or bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0692] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 244 as residues: Ser-35 to Phe-41.
[0693] The tissue distribution indicates that polynucleotides and
polypeptides corresponding to this gene are useful for study,
treatment, and diagnosis of disorders of the meningima. The protein
is also useful in the development of inhibitors of infections,
particularly, though not limited to, the meninges or other
neural-associated or neural tissue. In addition, the protein is
useful for the treatment of injuries to the meninges, potentially
in regeneration, or in congenital disorders, birth defects, etc.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0694] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:105 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 544 of SEQ ID NO: 105, b is an
integer of 15 to 558, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO: 105, and where
b is greater than or equal to a+14.
[0695] Features of Protein Encoded by Gene No: 96
[0696] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
40 MSTGDGRDAEKGWPVSEEENQRSVYPGYPECDERQAVPQHCAIASPSSLQSHH, (SEQ ID
NO:558) PASACVPRR QQMTLGTKIKWGQLQRGQEIPTGDFTVRNFM- , (SEQ ID
NO:559) RFSIIYC and/or PFLFCASRIRXQGIGIHGQVACSAVRMYNNR (SEQ ID
NO:560).
[0697] Polynucleotides encoding these polypeptides are also
encompassed by the invention. The gene encoding the disclosed cDNA
is thought to reside on chromosome 10. Accordingly, polynucleotides
related to this invention are useful as a marker in linkage
analysis for chromosome 10.
[0698] This gene is expressed primarily in neutrophils and
activated monocytes.
[0699] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune and hematopoietic disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
and hematopoietic systems, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., immune, cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0700] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 245 as residues: Met-l to Ser-6, Pro-29 to
Ser-34.
[0701] The tissue distribution in monocytes and neutrophils
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the study, diagnosis and treatment of
diseases of the immune and hematopoietic systems. Expression of
this gene product in monocytes and neutrophils also strongly
indicates a role for this protein in immune function and immune
surveillance. This gene product may be involved in the regulation
of cytokine production, antigen presentation, or other processes
that may also suggest a usefulness in the treatment of cancer (e.g.
by boosting immune responses). Since the gene is expressed in cells
of lymphoid origin, the gene or protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues. Protein,
as well as, antibodies directed against the protein may show
utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0702] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 106 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 742 of SEQ ID NO: 106, b is an
integer of 15 to 756, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO: 106, and where
b is greater than or equal to a+14.
[0703] Features of Protein Encoded by Gene No: 97
[0704] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
41 VLCEEAGQKVPSTPSWSSWTLQKRLRGSPAEANCSPSFPAPPGKE (SEQ ID NO:561),
MSLSALACDFTPIQPWEWEEYEQITLGLTAPSNLLESNYLGQASE (SEQ ID NO:562),
CFVRKLVRRFPQLLPGPPGHCRKDLGDPQQRPIALLPSLPHQERNNVHRLEAD SEVDL
CVDFDEYFSSWEPLLKMMFKGVVGGKMKAW (SEQ ID NO:563), and/or
RRKKRRKPLPYKIHAD MMFKGVVGGKMKAWRR (SEQ ID NO:564).
KKRRKPLPYKIHAD
[0705] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0706] This gene is expressed primarily in bone marrow, and to a
lesser extent in testes.
[0707] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, hematopoietic and reproductive disorders. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
hematopoietic and reproductive systems, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., reproductive, immune,
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0708] The tissue distribution in bone marrow and testes indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for the study, diagnosis and treatment of various
disorders involving the hematopoietic and reproductive systems. The
uses include bone marrow cell ex vivo culture, bone marrow
transplantation, bone marrow reconstitution, radiotherapy or
chemotherapy of neoplasia. The gene product may also be involved in
lymphopoiesis, therefore, it can be used in immune disorders such
as infection, inflammation, allergy, immunodeficiency etc. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types. Furthermore, polynucleotides and polypeptides
corresponding to this gene are useful for the treatment and
diagnosis of conditions concerning proper testicular function (e.g.
endocrine function, sperm maturation), as well as cancer.
Therefore, this gene product is useful in the treatment of male
infertility and/or impotence. This gene product is also useful in
assays designed to identify binding agents, as such agents
(antagonists) are useful as male contraceptive agents.
[0709] Similarly, the protein is believed to be useful in the
treatment and/or diagnosis of testicular cancer. The testes are
also a site of active gene expression of transcripts that may be
expressed, particularly at low levels, in other tissues of the
body. Therefore, this gene product may be expressed in other
specific tissues or organs where it may play related functional
roles in other processes, such as hematopoiesis, inflammation, bone
formation, and kidney function, to name a few possible target
indications. Protein, as well as, antibodies directed against the
protein may show utility as a tissue-specific marker and/or
immunotherapy target for the above listed tissues.
[0710] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:107 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1132 of SEQ ID NO:107, b is an
integer of 15 to 1146, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO:107, and where
b is greater than or equal to a+14.
[0711] Features of Protein Encoded by Gene No: 98
[0712] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
42 LISSVNKTKQKRSDATLSHKHDRLLNHFVFFGNSYNY (SEQ ID NO:565), SSK
FPSDMLLRIQQIIYCHKLTIILTKWRNTARHKSKKKEDELILKHELQLKKWKNR (SEQ ID
NO:566), LILKRAAAEESNFPERSSSEVFLVDETLKCDISLLPEXAILQVCMNSVY- IIYYNLP
SVVVHACNPSCLGG SLESTNAIKSN (SEQ ID NO:567), IRP (SEQ ID NO:568)
NKNDQMRHCLINMIDY ITLCFLETAITINIYSNLVNFLQICYC (SEQ ID NO:569),
and/or GYNRSSIVTS ISFRYAIADTTDHLLSQANHYPNKMA (SEQ ID NO:570).
EYSKT
[0713] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0714] This gene is expressed primarily in T-cells, tonsils, and
heart tissue.
[0715] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of immune system and vascular tissue disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune system and vascular tissue, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, vascular, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0716] The tissue distribution in T-cells, tonsils and heart
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the diagnosis and treatment of disorders
of the immune system and vascular tissues. Expression of this gene
product in tonsils indicates a role in the regulation of the
proliferation; survival; differentiation; and/or activation of
potentially all hematopoietic cell lineages, including blood stem
cells. This gene product may be involved in the regulation of
cytokine production, antigen presentation, or other processes that
may also suggest a usefulness in the treatment of cancer (e.g. by
boosting immune responses). Since the gene is expressed in cells of
lymphoid origin, the gene or protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
Therefore it may be also used as an agent for immunological
disorders including arthritis, asthma, immune deficiency diseases
such as AIDS, leukemia, rheumatoid arthritis, inflammatory bowel
disease, sepsis, acne, and psoriasis. In addition, this gene
product may have commercial utility in the expansion of stem cells
and committed progenitors of various blood lineages, and in the
differentiation and/or proliferation of various cell types.
[0717] Expression of this gene product in T cells also strongly
indicates a role for this protein in immune function and immune
surveillance. The tissue distribution in heart muscle tissue
indicates that the protein product of this gene is useful for the
diagnosis and treatment of conditions and pathologies of the
cardiovascular system, such as heart disease, restenosis,
atherosclerosis, stoke, angina, thrombosis, and wound healing.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0718] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:108 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 761 of SEQ ID NO: 108, b is an
integer of 15 to 775, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO: 108, and where
b is greater than or equal to a+14.
[0719] Features of Protein Encoded by Gene No: 99
[0720] An embodiment of the invention is directed to polypeptides
comprising those which exhibit sequence homology with honeybee
venom sacepin. In specific embodiments, polypeptides of the
invention comprise the following amino acid sequence:
43 PQIKLLNSDALGMRTTSXDLVPCNQCFIPLPPSCNRIASRKAVNWKQQRLPAVR (SEQ ID
NO:571), GLLNNAPHRRPPTPRTPCVFPSEGPKGYGFHV EQLAXISCR (SEQ ID
NO:572), VINVSFRCLHHVIESLPERQLTGSSRGSQP EDCSTMPPIAAP (SEQ ID
NO:573), and/or PPLAPLVFSPLRGPRVMAFMSRCGDRGGRGRSXAGRGWPWSESGVINAHPK
KRPCPGPMLS EFGTRRQWGTRCFPPLVGRKQSALR (SEQ ID NO:574).
RREGKARAGRCCGKRSVKAGFDA
[0721] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0722] This gene is expressed primarily in activated and control
neutrophils, and to a lesser extent in fetal liver and spleen.
[0723] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of disorders of the immune and endocrine systems.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune, inflammatory and hormonal systems, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0724] The tissue distribution in neutrophils and fetal liver and
spleen indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the study, diagnosis and
treatment of inflammation and various disorders of the immune and
endocrine systems. This gene product may be involved in the
regulation of cytokine production, antigen presentation, or other
processes that may also suggest a usefulness in the treatment of
cancer (e.g. by boosting immune responses). Since the gene is
expressed in cells of lymphoid origin, the gene or protein, as well
as, antibodies directed against the protein may show utility as a
tumor marker and/or immunotherapy targets for the above listed
tissues. Therefore it may be also used as an agent for
immunological disorders including arthritis, asthma, immune
deficiency diseases such as AIDS, leukemia, rheumatoid arthritis,
inflammatory bowel disease, sepsis, acne, and psoriasis.
[0725] In addition, this gene product may have commercial utility
in the expansion of stem cells and committed progenitors of various
blood lineages, and in the differentiation and/or proliferation of
various cell types. Expression of this gene product in neutrophils
also strongly indicates a role for this protein in immune function
and immune surveillance. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0726] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:109 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 897 of SEQ ID NO:109, b is an integer
of 15 to 911, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:109, and where b is greater
than or equal to a+14.
[0727] Features of Protein Encoded by Gene No: 100
[0728] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
44 AFFLLQALEIQSQLATPASSTARNPAPDLHHPHQPTIERFCRHSSSWERIEY (SEQ ID
NO:575).
[0729] Polynucleotides encoding these polypeptides are also
encompassed by the invention. When tested against Jurkat T-cell
lines, supernatants removed from cells containing this gene
activated the GAS assay. Thus, it is likely that this gene
activates T-cells through the Jak-STAT signal transduction pathway.
The gamma activating sequence (GAS) is a promoter element found
upstream of many genes which are involved in the Jak-STAT pathway.
The Jak-STAT pathway is a large, signal transduction pathway
involved in the differentiation and proliferation of cells.
Therefore, activation of the Jak-STAT pathway, reflected by the
binding of the GAS element, can be used to indicate proteins
involved in the proliferation and differentiation of cells.
Furthermore, contact of cells with supernatant expressing the
product of this gene increases the permeability of Brian
microvascular pericyte cells to calcium. Thus, it is likely that
the product of this gene is involved in a signal transduction
pathway that is initiated when the product of this gene binds a
receptor on the surface of the pericyte cell. Thus, polynucleotides
and polypeptides have uses which include, but are not limited to,
activating pericyte (endothelial) cells.
[0730] This gene is expressed primarily in activated
neutrophils.
[0731] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune disorders, inflammation. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard-gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0732] The tissue distribution in neutrophils and the biological
activity data suggest that polynucleotides and polypeptides
corresponding to this gene are useful for the study and treatment
of inflammatory and immune conditions. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer (e.g. by boosting immune responses).
Since the gene is expressed in cells of lymphoid origin, the gene
or protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues. Therefore it may be also used as an agent for
immunological disorders including arthritis, asthma, immune
deficiency diseases such as AIDS, leukemia, rheumatoid arthritis,
inflammatory bowel disease, sepsis, acne, and psoriasis.
[0733] In addition, this gene product may have commercial utility
in the expansion of stem cells and committed progenitors of various
blood lineages, and in the differentiation and/or proliferation of
various cell types. Expression of this gene product in neutrophils
also strongly indicates a role for this protein in immune function
and immune surveillance. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0734] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:110 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 442 of SEQ ID NO:110, b is an integer
of 15 to 456, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:110, and where b is greater
than or equal to a+14.
[0735] Features of Protein Encoded by Gene No: 101
[0736] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
45 ATVPGSIYNYFYHYNAGALKPEHASESPRGLCAQTAGPFPSF (SEQ ID NO:576),
IRHEPPPPRFKRFSCLSLLSSWDYRRAPPHVAIFCTLSRDGVLPHWPGWSQTPDLK (SEQ ID
NO:577), STHLGLPRCWDYRHEPLCLAPFTTISIIIMQGLSNLSM-
PQNPPEGCAHRLLDLSPASDSVPPEWGSKIAFEV (SEQ ID NO:578), and/or
LRVGGTSENCCRGECCGSVCIPPGRL (SEQ ID NO:579).
[0737] Polynucleotides encoding these polypeptides are also
encompassed by the invention. When tested against Jurkat T-cell
lines, supernatants removed from cells containing this gene
activated the GAS assay. Thus, it is likely that this gene
activates T-cells through the Jak-STAT signal transduction pathway.
The gamma activating sequence (GAS) is a promoter element found
upstream of many genes which are involved in the Jak-STAT pathway.
The Jak-STAT pathway is a large, signal transduction pathway
involved in the differentiation and proliferation of cells.
Therefore, activation of the Jak-STAT pathway, reflected by the
binding of the GAS element, can be used to indicate proteins
involved in the proliferation and differentiation of cells.
[0738] This gene is expressed primarily in neutrophils.
[0739] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune disorders. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0740] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study and treatment of immune disorders. Expression
of this gene product in neutrophils also strongly indicates a role
for this protein in immune function and immune surveillance. This
gene product may be involved in the regulation of cytokine
production, antigen presentation, or other processes that may also
suggest a usefulness in the treatment of cancer (e.g. by boosting
immune responses).
[0741] Since the gene is expressed in cells of lymphoid origin, the
gene or protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues. Therefore it may be also used
as an agent for immunological disorders including arthritis,
asthma, immune deficiency diseases such as AIDS, leukemia,
rheumatoid arthritis, inflammatory bowel disease, sepsis, acne, and
psoriasis. In addition, this gene product may have commercial
utility in the expansion of stem cells and committed progenitors of
various blood lineages, and in the differentiation and/or
proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0742] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:111 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 540 of SEQ ID NO:111, b is an integer
of 15 to 554, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:111, and where b is greater
than or equal to a+14.
[0743] Features of Protein Encoded by Gene No: 102
[0744] This gene is expressed primarily in neutrophils.
[0745] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune conditions. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0746] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 251 as residues: Lys-33 to Lys-41.
[0747] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for study and treatment of immune disorders. This gene
product may be involved in the regulation of cytokine production,
antigen presentation, or other processes that may also suggest a
usefulness in the treatment of cancer (e.g. by boosting immune
responses). Since the gene is expressed in cells of lymphoid
origin, the gene or protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues. Therefore it
may be also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia, rheumatoid arthritis, inflammatory bowel disease, sepsis,
acne, and psoriasis. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0748] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 112 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 708 of SEQ ID NO:112, b is an integer
of 15 to 722, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:112, and where b is greater
than or equal to a+14.
[0749] Features of Protein Encoded by Gene No: 103
[0750] When tested against K562 leukemia cell lines, supernatants
removed from cells containing this gene activated the ISRE assay.
Thus, it is likely that this gene activates leukemia cells through
the Jak-STAT signal transduction pathway. The interferon-sensitive
response element is a promoter element found upstream of many genes
which are involved in the Jak-STAT pathway. The Jak-STAT pathway is
a large, signal transduction pathway involved in the
differentiation and proliferation of cells. Therefore, activation
of the Jak-STAT pathway, reflected by the binding of the ISRE
element, can be used to indicate proteins involved in the
proliferation and differentiation of cells. In specific
embodiments, polypeptides of the invention comprise the following
amino acid sequence:
46 MCVTRMHVKCPPPSASVTAVKWPLSWSSSSFCISLHAGRH (SEQ ID NO:580), and/or
EERNKNHLSCQGLSTICCSYLSSKGEHLRNLSPYSF (SEQ ID NO:581).
[0751] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0752] This gene is expressed primarily in neutrophils.
[0753] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, immune conditions. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the immune system, expression of
this gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0754] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study and treatment of immune disorders. This gene
product may be involved in the regulation of cytokine production,
antigen presentation, or other processes that may also suggest a
usefulness in the treatment of cancer (e.g. by boosting immune
responses). Since the gene is expressed in cells of lymphoid
origin, the gene or protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues. Therefore it
may be also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia, rheumatoid arthritis, inflammatory bowel disease, sepsis,
acne, and psoriasis. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Expression of this gene
product in neutrophils also strongly indicates a role for this
protein in immune function and immune surveillance. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0755] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:113 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 917 of SEQ ID NO:113, b is an integer
of 15 to 931, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:113, and where b is greater
than or equal to a+14.
[0756] Features of Protein Encoded by Gene No: 104
[0757] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
47 GLCMVHSLLTSSLGGRCCNYPYIADKDIETEVKPPSQGHTWHLHCS (SEQ ID NO:582),
QLWCITALPSTRHCSKGFAWFTHSLRHPSVAGAVIILILQTRTLRQRSSHLPKGTHG-
ICTAPDRPTERAAVTILK (SEQ ID NO:583),
SFDNNNSYGVSQLYQVPDTVLRALHGSLTPYVIPRWQVL (SEQ ID NO:584), and/or
DRGQATFPRAHMASALLLTDRQRELLSRSSNELCMSKV (SEQ ID NO:585).
[0758] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0759] This gene is expressed primarily in neutrophils.
[0760] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of immune diseases and disorders. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
immune, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0761] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study and treatment of immune disorders. This gene
product may be involved in the regulation of cytokine production,
antigen presentation, or other processes that may also suggest a
usefulness in the treatment of cancer (e.g. by boosting immune
responses). Since the gene is expressed in cells of lymphoid
origin, the gene or protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues. Therefore it
may be also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia, rheumatoid arthritis, inflammatory bowel disease, sepsis,
acne, and psoriasis. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Expression of this gene
product in neutrophils also strongly indicates a role for this
protein in immune function and immune surveillance. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0762] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:114 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 574 of SEQ ID NO:114, b is an integer
of 15 to 588, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO: 114, and where b is greater
than or equal to a+14.
[0763] Features of Protein Encoded by Gene No: 105
[0764] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
48
LLLILRPFLNSQFKLQLPLVLFHSSCTYICLLYNYELFHIVALTGKLMNLGLHLFAHHLILAVA-
HXGCSIPIY (SEQ ID NO:586), and/or THNSNYSSLWFSSTAVVLTYVYY-
IIMNCFILSPLQVN (SEQ ID NO:587).
[0765] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0766] This gene is expressed primarily in neutrophils.
[0767] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of immune disorders and diseases. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
immune, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0768] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study and treatment of inflammatory and immune
disorders. This gene product may be involved in the regulation of
cytokine production, antigen presentation, or other processes that
may also suggest a usefulness in the treatment of cancer (e.g. by
boosting immune responses). Since the gene is expressed in cells of
lymphoid origin, the gene or protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
Therefore it may be also used as an agent for immunological
disorders including arthritis, asthma, immune deficiency diseases
such as AIDS, leukemia, rheumatoid arthritis, inflammatory bowel
disease, sepsis, acne, and psoriasis. In addition, this gene
product may have commercial utility in the expansion of stem cells
and committed progenitors of various blood lineages, and in the
differentiation and/or proliferation of various cell types.
Expression of this gene product in neutrophils also strongly
indicates a role for this protein in immune function and immune
surveillance. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0769] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:115 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 798 of SEQ ID NO:115, b is an integer
of 15 to 812, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:115, and where b is greater
than or equal to a+14.
[0770] Features of Protein Encoded by Gene No: 106
[0771] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
49 TLVAGSPCSLSRWIMAGFCHGELVQSDMESQEWERGQVVLSHTSLPWCYVSP (SEQ ID
NO:588), R MAGFCHGELVQSDMESQEWERGQVVLSHTSLPWCYVSP (SEQ ID NO:589),
R MAVWISGSYSSFCSRSNWDVFSPNIVLASLP- FSFRSVSK (SEQ ID NO:590), and/or
AAKPWWLALPALFPDGLWLDSAMGSLYSQTWKAR- NGKEVRWFSPTPHCLGA MSHL
GWLYGSVGLIPHSAAEATG (SEQ ID NO:591).
[0772] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0773] This gene is expressed primarily in neutrophils.
[0774] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of immune disorders and diseases. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
immune, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0775] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 255 as residues: Pro-54 to Gly-62.
[0776] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study and treatment of immune disorders. This gene
product may be involved in the regulation of cytokine production,
antigen presentation, or other processes that may also suggest a
usefulness in the treatment of cancer (e.g. by boosting immune
responses). Since the gene is expressed in cells of lymphoid
origin, the gene or protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues. Therefore it
may be also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia, rheumatoid arthritis, inflammatory bowel disease, sepsis,
acne, and psoriasis. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Expression of this gene
product in neutrophils also strongly indicates a role for this
protein in immune function and immune surveillance. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0777] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:116 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 492 of SEQ ID NO:116, b is an integer
of 15 to 506, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:116, and where b is greater
than or equal to a+14.
[0778] Features of Protein Encoded by Gene No: 107
[0779] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
50 RSKRQSQGSRCSVPLLAQQSRSPPVPLQAQPAWLLGSETIAWSGGGSGWEGPR (SEQ ID
NO:592), DPGTSTAAGNSGPGIGMGHRTPPPSHTGR RWDPAWGLDIP (SEQ ID NO:593),
ESSCPVTMGELRSGDGIVL GALLWDNSMISAPRGSHREA (SEQ ID NO:594),
GALFPSWLSNPAVLPSRSRPSQPGCLDP- RQ NSAREPRRWIR (SEQ ID NO:595),
PTRGSGETTAPCCFEPLNGGTLVHAAAMARASEAAGTG MARASEAAGTG (SEQ ID NO:596),
CFTTAFQKALRDPRPTLPDTHGSLRNAP (SEQ ID NO:597),
LKSLTLPAAFVVSFFFLSLLQDGIKERSQTQNATFFFHDRSDIEGLSEEPCSGTTP
LALQEAVTGKQVLCSPPGSAIPQSSRPAPGPASLAAWIRDN (SEQ ID NO:598), and/or
SLVWRRLRVGGTQGPGHQYSSWEFRPRDRDGAQDTTPISHREMKVGSSMGTG HP
MAGRLFTLLLWQELARRLVPGDASPRLSRKR (SEQ ID NO:599).
SVTPGPPFPTLTVPSE
[0780] Polynucleotides encoding these polypeptides are also
encompassed by the invention. When tested against sensory neuron
cell lines, supernatants removed from cells containing this gene
activated the EGR1 assay. Thus, it is likely that this gene
activates sensory neuron cells through a signal transduction
pathway. Early growth response 1 (EGR1) is a promoter associated
with certain genes that induces various tissues and cell types upon
activation, leading the cells to undergo differentiation and
proliferation.
[0781] This gene is expressed primarily in neutrophils.
[0782] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of immune disorders and diseases. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the immune system,
expression of this gene at significantly higher or lower levels may
be routinely detected in certain tissues or cell types (e.g.,
immune, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0783] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 256 as residues: Met-25 to Gly-30.
[0784] The tissue distribution in neutrophils indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study and treatment of immune disorders. This gene
product may be involved in the regulation of cytokine production,
antigen presentation, or other processes that may also suggest a
usefulness in the treatment of cancer (e.g. by boosting immune
responses). Since the gene is expressed in cells of lymphoid
origin, the gene or protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues. Therefore it
may be also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia, rheumatoid arthritis, inflammatory bowel disease, sepsis,
acne, and psoriasis. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Expression of this gene
product in neutrophils also strongly indicates a role for this
protein in immune function and immune surveillance. Protein, as
well as, antibodies directed against the protein may show utility
as a tumor marker and/or immunotherapy targets for the above listed
tissues.
[0785] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:117 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 737 of SEQ ID NO:117, b is an integer
of 15 to 751, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:117, and where b is greater
than or equal to a+14.
[0786] Features of Protein Encoded by Gene No: 108
[0787] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
51 MFYSKIFYELLLNSDTSNNVTSKTLVSSISSSNNRLAVSIVF (SEQ ID NO:600),
SRQKNLLKLHSNPNCDNFCFIFNYKPKYICIFKLICLKILLYIFGSG (SEQ ID NO:601),
and/or MLLSLLMVFTSELYVKRHISFKSXDKPHCHKNQDIDVLFRK- LLEKHFKVINMICFP
(SEQ ID NO:602).
[0788] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0789] This gene is expressed primarily in fetal liver, and to a
lesser extent in bone and breast cancer cell lines.
[0790] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, cancer and metabolic disorders. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the digestive and skeletal
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., digestive, skeletal, cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0791] The tissue distribution in fetal liver/spleen, as well as
bone and breast cancer, indicates that polynucleotides and
polypeptides corresponding to this gene are useful for study and
treatment of growth and metabolic disorders and neoplasias (e.g.
hepatoblastoma, jaundice, hepatitis, liver metabolic diseases and
conditions that are attributable to the differentiation of
hepatocyte progenitor cells). Furthermore, the tissue distribution
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the diagnosis and/or treatment of bone and
breast cancer, as well as cancers of other tissues where expression
has been observed. The expression within fetal tissue and other
cellular sources marked by proliferating cells suggests this
protein may play a role in the regulation of cellular division, and
may show utility in the diagnosis and treatment of cancer and other
proliferative disorders. Similarly, developmental tissues rely on
decisions involving cell differentiation and/or apoptosis in
pattern formation. Thus this protein may also be involved in
apoptosis or tissue differentiation and could again be useful in
cancer therapy. The protein product of this clone is useful for the
treatment and diagnosis of hematopoietic related disorders such as
anemia, pancytopenia, leukopenia, thrombocytopenia or leukemia
since stromal cells are important in the production of cells of
hematopoietic lineages. The uses include bone marrow cell ex-vivo
culture, bone marrow transplantation, bone marrow reconstitution,
radiotherapy or chemotherapy of neoplasia. The gene product may
also be involved in lymphopoiesis, therefore, it can be used in
immune disorders such as infection, inflammation, allergy,
immunodeficiency etc. In addition, this gene product may have
commercial utility in the expansion of stem cells and committed
progenitors of various blood lineages, and in the differentiation
and/or proliferation of various cell types. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0792] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:118 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 946 of SEQ ID NO:118, b is an integer
of 15 to 960, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:118, and where b is greater
than or equal to a+14.
[0793] Features of Protein Encoded by Gene No: 109
[0794] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
52 FREYGFYNLHFC (SEQ ID NO:603),
LVTTDYYDGCNEDYEYNWSYMFLNSEQLFIKFYPTEFC (SEQ ID NO:604), and/or
NVIAPGLESSCANSLFLLFVCLPVAHHRHNFLFIKHSLYNHLRDYESDFDKI (SEQ ID
NO:605).
[0795] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0796] This gene is expressed primarily in T cells, fetal heart and
infant brain, and to a lesser extent in some transformed cell
lines.
[0797] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of growth and immune disorders and diseases. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the immune
and cardiovascular systems, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, cardiovascular,
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine; synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0798] The tissue distribution in immune cells, heart tissue, and
brain tissue indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the study and treatment
of developmental and immune disorders. This gene product may be
involved in the regulation of cytokine production, antigen
presentation, or other processes that may also suggest a usefulness
in the treatment of cancer (e.g. by boosting immune responses).
Since the gene is expressed in cells of lymphoid origin, the gene
or protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues. Therefore it may be also used as an agent for
immunological disorders including arthritis, asthma, immune
deficiency diseases such as AIDS, leukemia, rheumatoid arthritis,
inflammatory bowel disease, sepsis, acne, and psoriasis. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types. Expression of this gene product in T cells also
strongly indicates a role for this protein in immune function and
immune surveillance.
[0799] The tissue distribution in heart muscle tissue indicates
that the protein product of this gene is useful for the diagnosis
and treatment of conditions and pathologies of the cardiovascular
system, such as heart disease, restenosis, atherosclerosis, stoke,
angina, thrombosis, and wound healing. The tissue distribution in
brain tissue indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the detection/treatment
of neurodegenerative disease states and behavioural disorders such
as Alzheimers Disease, Parkinsons Disease, Huntingtons Disease,
Tourette Syndrome, schizophrenia, mania, dementia, paranoia,
obsessive compulsive disorder, panic disorder, learning
disabilities, ALS, psychoses, autism, and altered bahaviors,
including disorders in feeding, sleep patterns, balance, and
perception. In addition, the gene or gene product may also play a
role in the treatment and/or detection of developmental disorders
associated with the developing embryo, or sexually-linked
disorders. Protein, as well as, antibodies directed against the
protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0800] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:119 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1428 of SEQ ID NO:119, b is an
integer of 15 to 1442, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO:1 19, and where
b is greater than or equal to a+14.
[0801] Features of Protein Encoded by Gene No: 110
[0802] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
53 PKVLAVLKKKNHVALSIFELLSNDICSFISFFMS (SEQ ID NO:606),
EGPDINSNLKELLCLKKKIMWPFQYLNC (SEQ ID NO:607), and/or
LLSLILLRIWYDFSKQTVFWFFLNVFNFFSSCNNDGACSYKYRKVQI (SEQ ID
NO:608).
[0803] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0804] This gene is expressed primarily in osteoblasts, and to a
lesser extent in bone marrow and bladder.
[0805] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, skeletal and hematopoietic diseases and disorders.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the skeletal and vascular systems, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., skeletal, vascular, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0806] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 259 as residues: Gly-33 to Lys-38.
[0807] The tissue distribution in bone marrow and osteoblasts
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the study, diagnosis, and treatment of
bone and hematopoietic disorders. Elevated levels of expression of
this gene product in osteoblasts indicates that it may play a role
in the survival, proliferation, and/or growth of osteoblasts.
Therefore, it may be useful in influencing bone mass in such
conditions as osteoporosis. More generally, as evidenced by
expression in bone marrow, this gene may play a role in the
survival, proliferation, and/or differentiation of hematopoietic
cells in general, and may be of use in augmentation of the numbers
of stem cells and committed progenitors. Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0808] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 120 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 831 of SEQ ID NO: 120, b is an
integer of 15 to 845, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO: 120, and where
b is greater than or equal to a+14.
[0809] Features of Protein Encoded by Gene No: 111
[0810] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
54 HTLFISFLWAEG (SEQ ID NO:609), MLPVFVLFFCFTYSARKQSVFKKGNVFE (SEQ
ID NO:610), and/or
SPCSAAECHNLSLLSSCSLVSSNILFSFPFFGQKARCCLFLFYFSASHIAHESRVYSKKEMCL
(SEQ ID NO:611).
[0811] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0812] This gene is expressed primarily in prostate cancer.
[0813] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, prostate and other cancers. Similarly, polypeptides and
antibodies directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the endocrine system, expression
of this gene at significantly higher or lower levels may be
routinely detected in certain tissues or cell types (e.g.,
endocrine, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0814] The tissue distribution in prostate cancer indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the study and treatment of prostate cancer, as well as
cancers of other tissues where expression has been observed.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0815] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:121 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 346 of SEQ ID NO:121, b is an integer
of 15 to 360, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:121, and where b is greater
than or equal to a+14.
[0816] Features of Protein Encoded by Gene No: 112
[0817] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
55 HKCFQCFILANGFLKVIKPFQRNWSDKTFFLVCLNKAISEALLSKMTFLSFFKT (SEQ ID
NO:612), NLLLLETFCTI LLGVLKPLYFSVEPVLGERSVAFEEVRE- KNH (SEQ ID
NO:613), GTSGFSLYSLAAIVCGHLMFFHTLLGRGGNDHPGQSPLPGMRPLRG- GLAGQ
APSGHPWMQPLDTCLL RPTRPPTRPDRPSLELAPGLCADF (SEQ ID NO:614), and/or
LGSSNHCIFLLSLYLGRDQ EKRIMVPQGFFPFTRWQP (SEQ ID NO:615).
LSVGTSCFSTLYWAVEVTITQASLLCLGCA- L
[0818] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0819] This gene is expressed primarily in haemopoietic and neural
tissues, and to a lesser extent in a number of cancers and other
tissues.
[0820] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the haemopoietic and neural systems.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune and neural system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, neural, cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0821] The tissue distribution in immune and neural tissues
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the diagnosis and treatment of diseases of
the haemopoietic and neural systems, including several cancers.
This gene product may be involved in the regulation of cytokine
production, antigen presentation, or other processes that may also
suggest a usefulness in the treatment of cancer (e.g. by boosting
immune responses). Since the gene is expressed in cells of lymphoid
origin, the gene or protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues. Therefore it
may be also used as an agent for immunological disorders including
arthritis, asthma, immune deficiency diseases such as AIDS,
leukemia, rheumatoid arthritis, inflammatory bowel disease, sepsis,
acne, and psoriasis.
[0822] In addition, this gene product may have commercial utility
in the expansion of stem cells and committed progenitors of various
blood lineages, and in the differentiation and/or proliferation of
various cell types. Furthermore, the tissue distribution indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses,
autism, and altered bahaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing embryo,
or sexually-linked disorders. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0823] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:122 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 930 of SEQ ID NO:122, b is an integer
of 15 to 944, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO: 122, and where b is greater
than or equal to a+14.
[0824] Features of Protein Encoded by Gene No: 113
[0825] The translation product of this gene shares sequence
homology with intestinal epithelium proliferating cell-associated
mRNA sequence, which is thought to be important in the growth and
development of epithelial cells. In specific embodiments,
polypeptides of the invention comprise the following amino acid
sequence:
56 MTLDEWKNLQEQTRPKPEFNIRKPESTVPSKAVVIRESKYRDDMVKDDYEDDS (SEQ ID
NO:616), HVFRKPANDITSQLEINFGNLPRPGRGARGGTRGGRGRIRRAENYGPRAEVVM
QDVAPNPDDPEDFPALS CKMLPPTQMTRKISLRCLERAL (SEQ ID NO:617),
FPSTAELHCTPVGRLFQLGQGSQTLRTIDVAFPVSCKFVALFWAELLEGLLQRL
ESRPFPKKMKNGDCVFIEGISNEE PPSSWAWSQRRHPG (SEQ ID NO:618), and/or
RPGKDQEGRELWTQSRSGDARCCPQPR CLKCVY (SEQ ID NO:619).
RDSIDSSAEAWRERRL
[0826] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0827] This gene is expressed primarily in brain and central
nervous system tissues, such as the frontal cortex, amygdala, and
hypothalmus, and to a lesser extent in testis.
[0828] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the neural system. Similarly, polypeptides
and antibodies directed to these polypeptides are useful in
providing immunological probes for differential identification of
the tissue(s) or cell type(s). For a number of disorders of the
above tissues or cells, particularly of the neural and reproductive
system, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., neural, reproductive, cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0829] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 262 as residues: Glu-20 to Glu-27, Glu-30 to
Trp-44.
[0830] The tissue distribution and homology to intestinal
epithelium proliferating cell-associated mRNA sequence indicates
that polynucleotides and polypeptides corresponding to this gene
are useful for growth and developmental diseases of the brain,
central nervous system and reproductive system. The tissue
distribution in neural tissues indicates that polynucleotides and
polypeptides corresponding to this gene are useful for the
detection/treatment of neurodegenerative disease states and
behavioural disorders such as Alzheimers Disease, Parkinsons
Disease, Huntingtons Disease, Tourette Syndrome, schizophrenia,
mania, dementia, paranoia, obsessive compulsive disorder, panic
disorder, learning disabilities, ALS, psychoses, autism, and
altered bahaviors, including disorders in feeding, sleep patterns,
balance, and perception.
[0831] In addition, the gene or gene product may also play a role
in the treatment and/or detection of developmental disorders
associated with the developing embryo, or sexually-linked
disorders. Elevated expression of this gene product within the
frontal cortex of the brain indicates that it may be involved in
neuronal survival; synapse formation; conductance; neural
differentiation, etc. Such involvement may impact many processes,
such as learning and cognition. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0832] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:123 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 900 of SEQ ID NO:123, b is an integer
of 15 to 914, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:123, and where b is greater
than or equal to a+14.
[0833] Features of Protein Encoded by Gene No: 114
[0834] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
57 LSYSVLLILPLFHSLPTLKDTHTHNKWVE (SEQ ID NO:620),
EVNGVGYKHSCFSDISSVLENKDSRMRAPHYASFQHFFSVLLKL (SEQ ID NO:621),
SPQACLTESQCIPLTFY KTHTHTISGWSKKSTELDISIPAFL (SEQ ID NO:622), and/or
TSPVSWRTRILE IRHELGSSDPPAEASQIAGTAAVS (SEQ ID NO:623). HHAQP
[0835] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0836] This gene is expressed primarily in spinal cord.
[0837] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, spinal cord injuries and diseases of the neural system.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the neural system, expression of this gene at significantly higher
or lower levels may be routinely detected in certain tissues or
cell types (e.g., neural, cancerous and wounded tissues) or bodily
fluids (e.g., lymph, serum, plasma, urine, synovial fluid and
spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0838] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 263 as residues: Pro-45 to Gln-52.
[0839] The tissue distribution in spinal cord indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis and treatment of spinal cord injuries and
diseases of the neural system, such as spinal deformation,
spinocerebullar ataxia types I and III, dentatorubropallidoluysian
and spinal bulbar muscular atrophy. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0840] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:124 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 448 of SEQ ID NO:124, b is an integer
of 15 to 462, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:124, and where b is greater
than or equal to a+14.
[0841] Features of Protein Encoded by Gene No: 115
[0842] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
58 MLYLILISLSSLSFSFSLPPFSIII (SEQ ID NO:624), SSYFL (SEQ ID
NO:625), RHFRIYHTCPKYFSMNIIN KLTLTKGNKSWSSTAVAAA (SEQ ID NO:626),
and/or LELVDPPGCRNSARDSLPNSTM MFYYACFILYSSLSPLSLSLSPSLLSLL
QFHTGNSYDHDYAK (SEQ ID NO:627).
[0843] Polynucleotides encoding these polypeptides are also
encompassed by the invention. When tested against U937 Myeloid cell
lines, supernatants removed from cells containing this gene
activated the GAS assay. Thus, it is likely that this gene
activates myeloid cells through the Jak-STAT signal transduction
pathway. The gamma activating sequence (GAS) is a promoter element
found upstream of many genes which are involved in the Jak-STAT
pathway. The Jak-STAT pathway is a large, signal transduction
pathway involved in the differentiation and proliferation of cells.
Therefore, activation of the Jak-STAT pathway, reflected by the
binding of the GAS element, can be used to indicate proteins
involved in the proliferation and differentiation of cells.
[0844] This gene is expressed primarily in striatum.
[0845] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of a number of diseases of the neural system. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the neural
diseases, expression of this gene at significantly higher or lower
levels may be routinely detected in certain tissues or cell types
(e.g., striatum, cancerous and wounded tissues) or bodily fluids
(e.g., lymph, serum, plasma, urine, synovial fluid and spinal
fluid) or another tissue or cell sample taken from an individual
having such a disorder, relative to the standard gene expression
level, i.e., the expression level in healthy tissue or bodily fluid
from an individual not having the disorder.
[0846] The tissue distribution in striatum indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis and treatment of diseases of the neural
system. Furthermore, the tissue distribution indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses,
autism, and altered bahaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing embryo,
or sexually-linked disorders. Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0847] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:125 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 531 of SEQ ID NO:125, b is an integer
of 15 to 545, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:125, and where b is greater
than or equal to a+14.
[0848] Features of Protein Encoded by Gene No: 116
[0849] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
59 AVCTGGYCESCRCEHCVCVCVDLCVLFSGKELRVR (SEQ ID NO:628),
VSFFFVFKWSFAEIKSREEHWASLTPKPTLLSALLTCDVLKS (SEQ ID NO:629),
SIIFKCCESTEDKGFDSFFQASKDGSSSRI RSWGSQRSLCLL (SEQ ID NO:630),
FIPFAAESYSVVWMGHLFVVCLLSSWWTFRPFALAVTVNHVAVNIVCVSAWTC
VSCSLGRSCGLEGSFLFPLETLWFPHMVVLCLTF MGHLFV (SEQ ID NO:631),
VCLLSSWWTFRPFALAVTVNHVAVNIVCVSAWTCVSCSLGRSCGLEGSFLFPL
ETLWFPHMVVLCLTF HDVLGARNAACVCCSFLLQQNRILL (SEQ ID NO:632),
FGWATCLLSVYSPAGGHLGRLHWRLL MLDFKTSQVSKAL (SEQ ID NO:633), and/or
KRVGFGVRLAQCSSLDLISAKLHLKTKKKETYITSTVMTAAS- LFLSYVTSEFTR
SIMATFYCFVLKLHIGEMGTLQTAGGSKMTWPLQKAIWQFLKRLSIKLPYVET RESPGETKNY
LTRNSFPENRTHKSTQTHTQCSQRHD (SEQ ID NO:634). SQ
[0850] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0851] This gene is expressed primarily in intestine and cancer
cells.
[0852] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the gastrointestinal tract and cancer.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the digestive system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., digestive, cancerous and wounded tissues) or
bodily fluids (e.g., lymph, serum, plasma, urine, synovial fluid
and spinal fluid) or another tissue or cell sample taken from an
individual having such a disorder, relative to the standard gene
expression level, i.e., the expression level in healthy tissue or
bodily fluid from an individual not having the disorder.
[0853] The tissue distribution in gastrointestinal tissues and
cancerous tissues indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the treatment and
diagnosis of diseases of the digestive system and cancer.
Furthermore, the tissue distribution in gastrointestinal tissues
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the diagnosis, prevention, and/or
treatment of various metabolic disorders such as Tay-Sachs disease,
phenylkenonuria, galactosemia, porphyrias, and Hurler's syndrome.
Protein, as well as, antibodies directed against the protein may
show utility as a tumor marker and/or immunotherapy targets for the
above listed tissues.
[0854] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:126 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 898 of SEQ ID NO:126, b is an integer
of 15 to 912, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO: 126, and where b is greater
than or equal to a+14.
[0855] Features of Protein Encoded by Gene No: 117
[0856] The translation product of this gene shares sequence
homology with a human apoptosis regulating protein which is thought
to be important in regulating cell death. In specific embodiments,
polypeptides of the invention comprise the following amino acid
sequence:
60 IRHEGQSSSRGSSHCDSPSPQEDGQIMFDVEMHTSRDHSSQSEEEVVEGEKEVE (SEQ ID
NO:635), ALKKSADWVSDWSSRPENIPPKEFHFRHPKRSVSLS
GILLTLYPFWPEDILEFPNRVYCCLEICKGFFSANATSRL (SEQ ID NO:636),
EFGTRDRVVPEAVLTVTALRHKKMGRSCLMWKCTPAGTIALSQKKKL (SEQ ID NO:637),
AHPLPAPTEGKEKPLEMRVTCEVVYCHSSLFELETIVSMTQPT (SEQ ID NO:638),
TLFLHIQFQ TFCVFKHEEKWSHEERGYFLRRISEGVHSISLP (SEQ ID NO:639), and/or
FSCFGFGARHLYWKATEHTLCQHLLRERKSPWKCV
QSLLLFRNLQGLLFRKCHQQIIILSAMLLSLISATRLDLYHSWYKFYSCNITTISL (SEQ ID
NO:640). LKRDQVSK
[0857] Polynucleotides encoding these polypeptides are also
encompassed by the invention. When tested against Jurkat cell
lines, supernatants removed from cells containing this gene
activated the NF-kB transcription factor. Thus, it is likely that
this gene activates Jurkat cells by activating a transcriptional
factor found within these cells. Nuclear factor kB is a
transcription factor activated by a wide variety of agents, leading
to cell activation, differentiation, or apoptosis. Reporter
constructs utilizing the NF-kB promoter element are used to screen
supernatants for such activity.
[0858] This gene is expressed primarily in muscle, fibroblast
cells, haemopoietic cells, and fetal lung.
[0859] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the haemopoietic, muscular and developing
systems. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune and muscular system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, muscular, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0860] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 266 as residues: Met-1 to Ala-6.
[0861] The tissue distribution in muscle and homology to apoptosis
regulating protein indicates that polynucleotides and polypeptides
corresponding to this gene are useful for diagnosis and treatment
of diseases of the haemopoietic, muscular and developing systems.
Expression within embryonic tissue and other cellular sources
marked by proliferating cells indicates that this protein may play
a role in the regulation-of cellular division. Additionally, the
expression in hematopoietic cells and tissues indicates that this
protein may play a role in the proliferation, differentiation,
and/or survival of hematopoietic cell lineages. In such an event,
this gene may be useful in the treatment of lymphoproliferative
disorders, and in the maintenance and differentiation of various
hematopoietic lineages from early hematopoietic stem and committed
progenitor cells. Similarly, embryonic development also involves
decisions involving cell differentiation and/or apoptosis in
pattern formation. Thus this protein may also be involved in
apoptosis or tissue differentiation and could again be useful in
cancer therapy. Protein, as well as, antibodies directed against
the protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0862] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:127 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1034 of SEQ ID NO:127, b is an
integer of 15 to 1048, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO:127, and where
b is greater than or equal to a+14.
[0863] Features of Protein Encoded by Gene No: 118
[0864] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
61 IRHEESFNPLTCGFSLFFSLFS, (SEQ ID NO: 641)
METLLLLLFFLSLLIFRFRILVSQCIN, (SEQ ID NO: 642)
FLLTTVLLFSSKVRDPRANFDQSLRVLKHAKKVQ (SEQ ID NO: 643)
PDVISKTSIMLGLGENDEQVYATMKGKEIEK, and/or QQSCCFPVRFVILGPILISPYVY.
(SEQ ID NO: 644)
[0865] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0866] This gene is expressed primarily in synovium.
[0867] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, arthritis and other diseases of the musculo-skeletal
system. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the musculo-skeletal system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., musculo-skeletal, cancerous
and wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0868] The tissue distribution in synovium indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the treatment and diagnosis of diseases of the
muscular-skeletal system. Furthermore, the expression of this gene
product in synovium indicates a role in the detection and treatment
of disorders and conditions affecting the skeletal system, in
particular osteoporosis as well as disorders afflicting connective
tissues (e.g. arthritis, trauma, tendonitis, chrondomalacia and
inflammation), such as in the diagnosis or treatment of various
autoimmune disorders such as rheumatoid arthritis, lupus,
scleroderma, and dermatomyositis as well as dwarfism, spinal
deformation, and specific joint abnormalities as well as
chondrodysplasias (ie. spondyloepiphyseal dysplasia congenita,
familial osteoarthritis, Atelosteogenesis type II, metaphyseal
chondrodysplasia type Schmid). Protein, as well as, antibodies
directed against the protein may show utility as a tumor marker
and/or immunotherapy targets for the above listed tissues.
[0869] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:128 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 708 of SEQ ID NO:128, b is an integer
of 15 to 722, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO: 128, and where b is greater
than or equal to a+14.
[0870] Features of Protein Encoded by Gene No: 119
[0871] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
62 VWLLSSILLRVLWNRYTLQELSFWLPWFASRATS (SEQ ID NO: 645)
LVLQHGDNYLLFLFCFVCFVLAMPF, IRHEVSMAFVFHLAQGTLEPLYIAGA, (SEQ ID NO:
646) NSARGEYGFCLPSCSGYFGTAIHCRSLASGYHGL (SEQ ID NO: 647) LPEQQA,
and/or HELTVPSRMGSKGKPYPCGFYSS- LIP. (SEQ ID NO: 648)
[0872] Polynucleotides encoding these polypeptides are also
encompassed by the invention. When tested against U937 Myeloid cell
lines, supernatants removed from cells containing this gene
activated the GAS assay. Thus, it is likely that this gene
activates myeloid cells through the Jak-STAT signal transduction
pathway. The gamma activating sequence (GAS) is a promoter element
found upstream of many genes which are involved in the Jak-STAT
pathway. The Jak-STAT pathway is a large, signal transduction
pathway involved in the differentiation and proliferation of cells.
Therefore, activation of the Jak-STAT pathway, reflected by the
binding of the GAS element, can be used to indicate proteins
involved in the proliferation and differentiation of cells.
[0873] This gene is expressed primarily in rejected kidney, stromal
cells, and infant brain.
[0874] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the renal, central nervous and immune
systems. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune, renal and central nervous system, expression of this
gene at significantly higher or lower levels may be routinely
detected in certain tissues or cell types (e.g., immune, renal,
nervous, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0875] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 268 as residues: Ser-6 to Arg-15.
[0876] The tissue distribution rejected kidney, stromal cells, and
infant brain indicates that polynucleotides and polypeptides
corresponding to this gene are useful for the diagnosis and
treatment of diseases of the renal, central nervous, and immune
systems. The tissue distribution in infant brain indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the detection/treatment of neurodegenerative disease
states and behavioural disorders such as Alzheimers Disease,
Parkinsons Disease, Huntingtons Disease, Tourette Syndrome,
schizophrenia, mania, dementia, paranoia, obsessive compulsive
disorder, panic disorder, learning disabilities, ALS, psychoses,
autism, and altered bahaviors, including disorders in feeding,
sleep patterns, balance, and perception. In addition, the gene or
gene product may also play a role in the treatment and/or detection
of developmental disorders associated with the developing embryo,
or sexually-linked disorders.
[0877] Alternatively, this gene product may be involved in the
regulation of cytokine production, antigen presentation, or other
processes that may also suggest a usefulness in the treatment of
cancer (e.g. by boosting immune responses). Since the gene is
expressed in cells of lymphoid origin, the gene or protein, as well
as, antibodies directed against the protein may show utility as a
tumor marker and/or immunotherapy targets for the above listed
tissues. Therefore it may be also used as an agent for
immunological disorders including arthritis, asthma, immune
deficiency diseases such as AIDS, leukemia, rheumatoid arthritis,
inflammatory bowel disease, sepsis, acne, and psoriasis. In
addition, this gene product may have commercial utility in the
expansion of stem cells and committed progenitors of various blood
lineages, and in the differentiation and/or proliferation of
various cell types.
[0878] The tissue distribution in kidney indicates that this gene
or gene product is useful in the treatment and/or detection of
kidney diseases including renal failure, nephritus, renal tubular
acidosis, proteinuria, pyuria, edema, pyelonephritis,
hydronephritis, nephrotic syndrome, crush syndrome,
glomerulonephritis, hematuria, renal colic and kidney stones, in
addition to Wilm's Tumor Disease, and congenital kidney
abnormalities such as horseshoe kidney, polycystic kidney, and
Falconi's syndrome. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0879] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:129 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 463 of SEQ ID NO:129, b is an integer
of 15 to 477, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:129, and where b is greater
than or equal to a+14.
[0880] Features of Protein Encoded by Gene No: 120
[0881] The protein of the invention has sequence identity to the
Saccharomyces cerevisiae ankyrin repeat-containing protein
(gi.vertline.466522). The translation product of this gene also
shares homology with C. elegans protein C43H6.7 gene product
(Genbank Accession No. gi.vertline.1255324). In specific
embodiments, polypeptides of the invention comprise the following
amino acid sequence:
63 KCIYPKPARTHHCSICNRCVLKMDHHCPWLNNC (SEQ ID NO: 649)
VGHYNHRYFFSFCFFMTLGCVYCSYGSWDLFREA YAAIEKMKQLDKNKLQAVANQTYHQTPPPTF-
SF RER, ARGHWNLILIVFHYYQAITTPPGYPPQGRNDIA (SEQ ID NO: 650) TVSIC,
WQCELDCVSHDSSTHSAPYVISRASKGSFSQNP, (SEQ ID NO: 651)
SKRASGPALGYHAGQFKDQPFYHCRRKTQCGEI (SEQ ID NO: 652)
LGLTSLYSGKQKFQPQTRGQAASYLPCPVLTRT SSRIQHWSWPPPLLLAV,
ESLQLRLLGQLEGIPGCGYRKALAYSGALTF, (SEQ ID NO: 653) and/or
SLAPWEWNELGAPSLGDCSLSLCDGS (SEQ ID NO: 654)
VSWTVSATTRALILLPMLFQGPPRAAFLRILDQ KEPVGLP.
[0882] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0883] This gene is expressed primarily in endometrial tumor, colon
tumor, prostate cancer, and ovarian cancer.
[0884] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of a number of types of cancers, particularly
endometrial, prostate, ovarian, and colon cancers. Similarly,
polypeptides and antibodies directed to these polypeptides are
useful in providing immunological probes for differential
identification of the tissue(s) or cell type(s). For a number of
disorders of the above tissues or cells, particularly of the
endometrium, prostate, colon, and ovary, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., prostate, colon, ovary,
endometrium, cancerous and wounded tissues) or bodily fluids (e.g.,
lymph, serum, plasma, urine, synovial fluid and spinal fluid) or
another tissue or cell sample taken from an individual having such
a disorder, relative to the standard gene expression level, i.e.,
the expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0885] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 269 as residues: Asn-43 to Arg-49, Phe-57 to Cys-65,
Pro-93 to Ser-99.
[0886] The tissue distribution in prostate cancer, ovarian cancer,
colon cancer, and endometrial cancer tissues indicates that
polynucleotides and polypeptides corresponding to this gene are
useful for the diagnosis and treatment of diseases and cancers of
the prostate, ovaries, colon, and endometrium, as well as cancers
of other tissues where expression has been observed. Expression
within cellular sources marked by proliferating cells suggests this
protein may play a role in the regulation of cellular division, and
may show utility in the diagnosis and treatment of cancer and other
proliferative disorders. Similarly, developmental tissues rely on
decisions involving cell differentiation and/or apoptosis in
pattern formation. Thus this protein may also be involved in
apoptosis or tissue differentiation and could again be useful in
cancer therapy. Protein, as well as, antibodies directed against
the protein may show utility as a tumor marker and/or immunotherapy
targets for the above listed tissues.
[0887] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:130 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 1282 of SEQ ID NO:130, b is an
integer of 15 to 1296, where both a and b correspond to the
positions of nucleotide residues shown in SEQ ID NO:130, and where
b is greater than or equal to a+14.
[0888] Features of Protein Encoded by Gene No: 121
[0889] The translation product of this gene shares sequence
homology with adrenalin receptor (Patent serial No. J08126491-A.)
In specific embodiments, polypeptides of the invention comprise the
following amino acid sequence:
64 TATLNSFFGGWGLALLLRLECSDTIMDHCSLDLL (SEQ ID NO: 655)
GSSNPPASASQVVGTTGARHHAQLIFCFFVQTRS HSVA,
MDHCSLDLLGSSNPPASASQVVGTTGARHHAQLI (SEQ ID NO: 656) FCFFVQTRSHSVA,
GVLKQSSHLVLSKG, (SEQ ID NO: 657) DYSCESLCPALLSIAPDIVLN, (SEQ ID NO:
658) TTIHKTQLGSYKILWEPKEGYHNSTWI, (SEQ ID NO: 659) IREIFLRRP,
and/or (SEQ ID NO: 660) LKFQKPGKIQMRGGGRVFWYKNCK. (SEQ ID NO:
661)
[0890] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0891] This gene is expressed primarily in synovial sarcoma.
[0892] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, arthritis and other diseases of the synovium including
cancers. Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the immune and muscular-skeletal system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., immune, musculo-skeletal,
cancerous and wounded tissues) or bodily fluids (e.g., lymph,
serum, plasma, urine, synovial fluid and spinal fluid) or another
tissue or cell sample taken from an individual having such a
disorder, relative to the standard gene expression level, i.e., the
expression level in healthy tissue or bodily fluid from an
individual not having the disorder.
[0893] The tissue distribution in synovial sarcoma tissue and the
homology to adrenalin receptor indicates that polynucleotides and
polypeptides corresponding to this gene are useful for the
diagnosis and treatment of diseases of the synovium, immune system
and musculo-skeletal system including cancers of these tissues and
systems. It may also be useful for identifying and therapeutically
using antagonists and agonists for this receptor family. In
addition, the expression of this gene product in synovium indicates
a role in the detection and treatment of disorders and conditions
affecting the skeletal system, in particular osteoporosis as well
as disorders afflicting connective tissues (e.g. arthritis, trauma,
tendonitis, chrondomalacia and inflammation), such as in the
diagnosis or treatment of various autoimmune disorders such as
rheumatoid arthritis, lupus, scleroderma, and dermatomyositis as
well as dwarfism, spinal deformation, and specific joint
abnormalities as well as chondrodysplasias (ie. spondyloepiphyseal
dysplasia congenita, familial osteoarthritis, Atelosteogenesis type
II, metaphyseal chondrodysplasia type Schmid). Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0894] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:131 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 724 of SEQ ID NO:131, b is an integer
of 15 to 738, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:131, and where b is greater
than or equal to a+14.
[0895] Features of Protein Encoded by Gene No: 122
[0896] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
65 NSARVTQKGESVGSVGCMRAIAGFDNYPLF, (SEQ ID NO: 662)
GTIGIFWPLPVAILSSGDYLQTQIHRPLLHRGT, (SEQ ID NO: 663)
LPLPLSSLLHIATCNPFPKT, (SEQ ID NO: 664)
SYFFVYNLILKIIQGDHASIILLATIPIFGDIYY (SEQ ID NO: 665) VKGQLASFGPYL,
LFYHLEIISRHKSIAHCSIEA, (SEQ ID NO: 666) CSCHCPSRAFST, and/or (SEQ
ID NO: 667) PHAIHSQKPSSIFLITDVFPDPPVGIYLL. (SEQ ID NO: 668)
[0897] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0898] This gene is expressed primarily in chronic synovitis.
[0899] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, inflammatory diseases -and disorders of the
musculo-skeletal system. Similarly, polypeptides and antibodies
directed to these polypeptides are useful in providing
immunological probes for differential identification of the
tissue(s) or cell type(s). For a number of disorders of the above
tissues or cells, particularly of the inflammatory and
musculo-skeletal system, expression of this gene at significantly
higher or lower levels may be routinely detected in certain tissues
or cell types (e.g., musculo-skeletal, cancerous and wounded
tissues) or bodily fluids (e.g., lymph, serum, plasma, urine,
synovial fluid and spinal fluid) or another tissue or cell sample
taken from an individual having such a disorder, relative to the
standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0900] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 271 as residues: Ser-39 to Pro-44.
[0901] The tissue distribution in chronic synovitis tissue
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the treatment and diagnosis of disorders
and diseases of the inflammatory and musculo-skeletal system. In
addition, the expression of this gene product in synovium indicates
a role in the detection and treatment of disorders and conditions
affecting the skeletal system, in particular osteoporosis as well
as disorders afflicting connective tissues (e.g. arthritis, trauma,
tendonitis, chrondomalacia and inflammation), such as in the
diagnosis or treatment of various autoimmune disorders such as
rheumatoid arthritis, lupus, scleroderma, and dermatomyositis as
well as dwarfism, spinal deformation, and specific joint
abnormalities as well as chondrodysplasias (ie. spondyloepiphyseal
dysplasia congenita, familial osteoarthritis, Atelosteogenesis type
II, metaphyseal chondrodysplasia type Schmid). Protein, as well as,
antibodies directed against the protein may show utility as a tumor
marker and/or immunotherapy targets for the above listed
tissues.
[0902] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO:132 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 428 of SEQ ID NO:132, b is an integer
of 15 to 442, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:132, and where b is greater
than or equal to a+14.
[0903] Features of Protein Encoded by Gene No: 123
[0904] In specific embodiments, polypeptides of the invention
comprise the following amino acid sequence:
66 RKLFHKINSKSFHLSGMHILISVWIVRSRIIKVK (SEQ ID NO: 669)
YELLLCFFDVIFYV, NSARDVFFTQKILYSQTCIFFPCLVPFSFLFSFF (SEQ ID NO: 670)
FFLSFVG, MFSSLKKFYILKHVYSFPVLFHFLFFFLF- SFSFL (SEQ ID NO: 671)
SWAEKGAGKMKLATENCKMVKS, and/or IQLLYLKGAAMKYLSYVARLLFLKALDLFAPKMV
(SEQ ID NO: 672) QIDSF.
[0905] Polynucleotides encoding these polypeptides are also
encompassed by the invention.
[0906] This gene is expressed primarily in kidney and infant
brain.
[0907] Therefore, polynucleotides and polypeptides of the invention
are useful as reagents for differential identification of the
tissue(s) or cell type(s) present in a biological sample and for
diagnosis of diseases and conditions which include, but are not
limited to, diseases of the renal and central nervous systems.
Similarly, polypeptides and antibodies directed to these
polypeptides are useful in providing immunological probes for
differential identification of the tissue(s) or cell type(s). For a
number of disorders of the above tissues or cells, particularly of
the neural and renal system, expression of this gene at
significantly higher or lower levels may be routinely detected in
certain tissues or cell types (e.g., neural, renal, cancerous and
wounded tissues) or bodily fluids (e.g., lymph, serum, plasma,
urine, synovial fluid and spinal fluid) or another tissue or cell
sample taken from an individual having such a disorder, relative to
the standard gene expression level, i.e., the expression level in
healthy tissue or bodily fluid from an individual not having the
disorder.
[0908] Preferred epitopes include those comprising a sequence shown
in SEQ ID NO. 272 as residues: Gly-24 to Lys-31.
[0909] The tissue distribution in kidney and infant brain tissues
indicates that polynucleotides and polypeptides corresponding to
this gene are useful for the diagnosis and treatment of diseases of
the neural and renal systems. The tissue distribution in infant
brain indicates that polynucleotides and polypeptides corresponding
to this gene are useful for the detection/treatment of
neurodegenerative disease states and behavioural disorders such as
Alzheimers Disease, Parkinsons Disease, Huntingtons Disease,
Tourette Syndrome, schizophrenia, mania, dementia, paranoia,
obsessive compulsive disorder, panic disorder, learning
disabilities, ALS, psychoses, autism, and altered bahaviors,
including disorders in feeding, sleep patterns, balance, and
perception. In addition, the gene or gene product may also play a
role in the treatment and/or detection of developmental disorders
associated with the developing embryo, or sexually-linked
disorders.
[0910] The tissue distribution in kidney indicates that this gene
or gene product is useful in the treatment and/or detection of
kidney diseases including renal failure, nephritus, renal tubular
acidosis, proteinuria, pyuria, edema, pyelonephritis,
hydronephritis, nephrotic syndrome, crush syndrome,
glomerulonephritis, hematuria, renal colic and kidney stones, in
addition to Wilm's Tumor Disease, and congenital kidney
abnormalities such as horseshoe kidney, polycystic kidney, and
Falconi's syndrome. Protein, as well as, antibodies directed
against the protein may show utility as a tumor marker and/or
immunotherapy targets for the above listed tissues.
[0911] Many polynucleotide sequences, such as EST sequences, are
publicly available and accessible through sequence databases. Some
of these sequences are related to SEQ ID NO: 133 and may have been
publicly available prior to conception of the present invention.
Preferably, such related polynucleotides are specifically excluded
from the scope of the present invention. To list every related
sequence would be cumbersome. Accordingly, preferably excluded from
the present invention are one or more polynucleotides comprising a
nucleotide sequence described by the general formula of a-b, where
a is any integer between 1 to 868 of SEQ ID NO:133, b is an integer
of 15 to 882, where both a and b correspond to the positions of
nucleotide residues shown in SEQ ID NO:133, and where b is greater
than or equal to a+14.
67 5' NT NT of AA First Last ATCC SEQ 5' NT 3' NT 5' NT First SEQ
AA AA First AA Last Deposit ID Total of of of AA of ID of of of AA
Gene cDNA Nr and NO. NT Clone Clone Start Signal NO: Sig Sig
Secreted of No. Clone ID Date Vector X Seq. Seq. Seq. Condon Pep Y
Pep Pep Portion ORF 1 HCEIA77 209119 Uni-ZAP XR 11 1882 676 1882
785 785 150 1 37 38 53 06/12/97 2 HCFCE10 209119 pSport1 12 1590 18
1590 198 198 151 1 19 20 45 06/12/97 3 HCFNC26 209119 pSport1 13
1373 6 1373 85 85 152 1 18 19 24 06/12/97 4 HCHAA63 209119 pSport1
14 1142 1 1142 130 130 153 1 37 38 264 06/12/97 5 HCNSP40 209119
pBluescript 15 1034 1 1034 106 106 154 1 19 20 237 06/12/97 5
HCNSP40 209119 pBluescript 134 1032 1 1032 111 273 1 14 06/12/97 6
HDAAC10 209119 pSport1 16 1198 1 1198 117 117 155 1 21 22 313
06/12/97 7 HE8CV18 209119 Uni-ZAP XR 17 1447 1 1447 176 176 156 1
29 30 98 06/12/97 8 HELDY05 209119 Uni-ZAP XR 18 1422 1 1375 79 79
157 1 34 35 36 06/12/97 9 HELDZ32 209119 Uni-ZAP XR 19 1107 12 1107
148 148 158 1 15 16 22 06/12/97 10 HFGAL10 209119 Uni-ZAP XR 20
1183 1 1183 179 179 159 1 20 21 96 06/12/97 10 HFGAL10 209119
Uni-ZAP XR 135 1766 3 1765 179 179 274 1 17 18 36 06/12/97 11
HFKEB72 209119 Uni-ZAP XR 21 1420 1 1420 43 43 160 1 29 30 65
06/12/97 12 HFTCU19 209119 Uni-ZAP XR 22 1575 1266 1575 137 137 161
1 30 31 222 06/12/97 12 HFTCU19 209119 Uni-ZAP XR 136 470 1 470 157
157 275 1 24 25 56 06/12/97 13 IIFXHN31 209119 Lambda ZAP 23 541 1
541 172 172 162 1 30 31 91 06/12/97 II 13 HFXHN31 209119 Lambda ZAP
137 1168 1 1168 293 293 276 1 22 23 26 06/12/97 II 14 HGLAM53
209119 Uni-ZAP XR 24 833 219 833 359 359 163 1 27 28 74 06/12/97 14
HGLAM53 209119 Uni-ZAP XR 138 1294 226 1288 369 277 1 26 27 67
06/12/97 15 HJABB94 209119 pBluescript 25 1555 1 1555 74 74 164 1
28 29 77 06/12/97 SK- 16 HKIYO61 209119 pBluescript 26 1543 1 1543
181 181 165 1 19 20 37 06/12/97 17 HLTAI94 209119 Uni-ZAP XR 27
1262 1 1262 47 47 166 1 18 19 44 06/12/97 18 HMDAI51 209119 Uni-ZAP
XR 28 753 1 753 12 12 167 1 21 22 38 06/12/97 19 HMELR03 209119
Lambda ZAP 29 1621 8 1535 200 200 168 1 25 26 173 06/12/97 II 20
HMKAH10 209119 pSport1 30 921 1 921 48 48 169 1 463 44 54 06/12/97
21 HMKCW19 209119 pSport1 31 2095 473 1934 529 529 170 1 30 31 344
06/12/97 21 HMKCW19 209119 pSport1 139 1720 103 1692 188 188 278 1
27 28 45 06/12/97 22 HMSJW18 209119 Uni-ZAP XR 32 1838 1 1838 28 28
171 1 23 24 89 06/12/97 23 HMWGY01 209119 Uni-Zap XR 33 782 1 782
423 423 172 1 30 31 104 06/12/97 23 HMWGY01 209119 Uni-Zap XR 140
774 1 774 17 17 279 1 31 32 39 06/12/97 24 HNFID82 209119
pBluescript 34 1560 1 1560 161 161 173 1 17 18 41 06/12/97 25
HNFIG36 209119 pBluescript 35 1092 1 1082 171 171 174 1 18 19 46
06/12/97 26 HNGEV29 209119 Uni-ZAP XR 36 1153 1 1153 173 173 175 1
30 31 73 06/12/97 26 HNGEV29 209119 Uni-ZAP XR 141 1566 1 1566 79
79 280 1 10 06/12/97 27 HNGIK21 209119 Uni-ZAP XR 37 985 1 985 152
152 176 1 25 26 28 06/12/97 28 HNGJJ65 209124 Uni-ZAP XR 38 1122 1
1122 84 84 177 1 22 23 67 06/19/97 29 HNGJU42 209124 Uni-ZAP XR 39
598 6 598 273 273 178 1 17 18 23 06/19/97 30 HODAZ26 209124 Uni-ZAP
XR 40 1129 8 1129 133 133 179 1 30 06/19/97 31 HODDB05 209124
Uni-ZAP XR 41 1158 22 1158 244 244 180 1 10 06/19/97 32 HOFAF39
209124 pSport1 42 1767 1 1767 57 57 181 1 22 23 31 06/19/97 33
HOFNY71 209124 pCMVSport 43 917 1 917 114 114 182 1 31 32 35
06/19/97 2.0 34 HORBI81 209124 Uni-ZAP XR 44 1987 8 1965 31 31 183
1 34 35 34 06/19/97 35 HOSCY73 209124 Uni-ZAP XR 45 2053 196 2048
209 209 184 1 28 06/19/97 36 HPMBR15 209124 Uni-ZAP XR 46 1272 25
1272 262 262 185 1 5 06/19/97 37 HSAVD46 209124 Uni-ZAP XR 47 773 2
767 155 155 186 1 20 21 58 06/19/97 38 HSLBF69 209124 Uni-ZAP XR 48
2119 1 2119 107 107 187 1 19 20 405 06/19/97 39 HSOAH66 209124
Uni-ZAP XR 49 1188 7 1188 196 196 188 1 27 28 36 06/19/97 39
HSOAH66 209124 Uni-ZAP XR 143 537 1 537 136 136 282 1 21 22 47
06/19/97 40 HSVBH58 209124 Uni-ZAP XR 50 478 24 155 249 249 189 1
40 41 57 06/19/97 40 HSVBH58 209124 Uni-ZAP XR 144 680 1 680 168
168 283 1 20 21 22 06/19/97 41 HSZAF47 209124 Uni-ZAP XR 51 1333 2
1333 107 107 190 1 18 19 126 06/19/97 42 HTADV27 209124 Uni-ZAP XR
52 1255 14 1255 69 69 191 1 20 21 20 06/19/97 43 HTADX17 209124
Uni-ZAP XR 53 1140 22 1140 84 84 192 1 24 25 142 06/19/97 44
HTDAD22 209124 pSport1 54 1220 1 1220 193 193 193 1 37 38 109
06/19/97 45 HTEDS39 209124 Uni-ZAP XR 55 694 198 694 205 205 194 1
21 22 80 06/19/97 45 HTEDS39 209124 Uni-ZAP XR 145 1048 1 1048 227
284 1 20 06/19/97 46 HTEHH53 209124 Uni-ZAP XR 56 988 1 980 22 22
195 1 24 25 209 06/19/97 47 HTLDP69 209124 Uni-ZAP XR 57 1500 237
1500 330 330 196 1 29 30 148 06/19/97 48 HTNBR95 209124 pBluescript
58 1391 1 1386 70 70 197 1 28 29 35 06/19/97 SK- 49 HTPCS60 209124
Uni-ZAP XR 59 1579 7 1259 105 105 198 1 19 20 257 06/19/97 50
HUKBH05 209124 Lambda ZAP 60 1241 1 1215 151 151 199 1 18 19 33
06/19/97 II 51 HUKEX85 209124 Lambda ZAP 61 930 7 925 35 35 200 1
18 19 33 06/19/97 II 51 HUKEX85 209124 Lambda ZAP 146 930 6 917 83
83 285 1 30 31 122 06/19/97 II 52 HWTBM45 209124 Uni-ZAP XR 62 998
1 998 69 69 201 1 19 20 25 06/19/97 53 HADFF38 209124 pSport1 63
1193 1 1034 64 64 202 1 19 20 33 06/19/97 54 HADFK68 209124 pSport1
64 830 1 830 91 91 203 1 24 25 58 06/19/97 54 HADFK68 209124
pSport1 147 830 1 830 45 45 286 1 26 27 26 06/19/97 55 HADGG19
209125 pSport1 65 867 1 867 262 262 204 1 30 31 75 06/19/97 55
IIADGG19 209125 pSport1 148 865 1 865 281 281 287 1 7 06/19/97 56
HAEAV45 209125 pBluescript 66 685 46 647 487 487 205 1 34 35 66
06/19/97 SK- 56 HAEAV45 209125 pBluescript 149 545 1 545 24 24 288
1 25 26 28 06/19/97 SK- 57 HARAA15 209125 pBluescript 67 801 1 801
185 185 206 1 34 35 43 06/19/97 SK- 58 HATDL27 209125 Uni-ZAP XR 68
908 1 908 82 82 207 1 28 29 31 06/19/97 59 HBAFQ54 209125 pSport1
69 696 209 696 229 229 208 1 20 21 47 06/19/97 60 HBGBA14 209125
Uni-ZAP XR 70 455 1 452 32 32 209 1 24 25 36 06/19/97 61 HBIAS26
209125 Uni-ZAP XR 71 413 1 372 57 57 210 1 27 28 73 06/19/97 62
HBJFU48 209125 Uni-ZAP XR 72 849 1 849 20 20 211 1 39 40 40
06/19/97 63 HBJFV28 209125 Uni-ZAP XR 73 505 1 505 306 306 212 1 21
22 53 06/19/97 64 HBMWB01 209125 Uni-ZAP XR 74 719 1 719 48 48 213
1 17 18 62 06/19/97 65 HBMXN79 209125 Uni-ZAP XR 75 1274 141 974
192 192 214 1 44 45 175 06/19/97 66 HBMXP84 209125 Uni-ZAP XR 76
519 1 519 161 161 215 1 31 32 39 06/19/97 67 HCFMM26 209125 pSport1
77 389 1 389 178 178 216 1 27 28 54 06/19/97 68 HCNAV36 209125
Lambda ZAP 78 823 411 823 505 505 217 1 15 16 46 06/19/97 II 69
HCNSB01 209125 pBluescript 79 2455 533 1308 552 552 218 1 22 23 179
06/19/97 70 HCRBR74 209125 Uni-ZAP XR 80 921 365 911 415 415 219 1
39 40 43 06/19/97 71 HCUBN59 209125 ZAP Express 81 678 1 678 96 96
220 1 39 40 43 06/19/97 72 HCUDB38 209125 ZAP Express 82 857 1 857
221 221 221 1 17 18 41 06/19/97 73 HCUFZ62 209125 ZAP Express 83
1977 28 661 233 233 222 1 28 29 51 06/19/97 74 HDHMB42 209125
pCMVSport 84 1149 427 1149 592 592 223 1 26 27 31 06/19/97 2.0 75
HDPCO25 209125 pCMVSport 85 767 76 767 182 182 224 1 20 21 53
06/19/97 3.0 76 HDPHI51 209125 pCMVSport 86 728 1 728 245 245 225 1
30 31 40 06/19/97 3.0 77 HE2EC79 209125 Uni-ZAP XR 87 735 1 735 151
151 226 1 21 22 30 06/19/97 78 HE9FE83 209125 Uni-ZAP XR 88 889 332
889 351 351 227 1 21 22 59 06/19/97 79 HE9HW52 209125 Uni-ZAP XR 89
569 73 569 122 122 228 1 25 26 34 06/19/97 80 HEBFL88 209125
Uni-ZAP XR 90 334 2 334 76 76 229 1 22 23 38 06/19/97 81 HFIVB57
209125 pSport1 91 795 92 795 286 286 230 1 35 36 38 06/19/97 82
HFPDE69 209125 Uni-ZAP XR 92 577 1 577 72 72 231 1 33 34 61
06/19/97 83 HGBGV89 209125 Uni-ZAP XR 93 968 1 968 55 55 232 1 26
27 197 06/19/97 84 HGLDE38 209125 Uni-ZAP XR 94 553 1 553 31 31 233
1 19 20 61 06/19/97 85 HHGDU58 209125 Lambda ZAP 95 968 70 898 235
235 234 1 46 47 80 06/19/97 II 86 HHTLF25 209125 ZAP Express 96 697
1 661 142 142 235 1 26 27 111 06/19/97 87 HJMAV91 209125 pCMVSport
97 866 74 866 251 251 236 1 16 17 32 06/19/97 3.0 88 HKAFB88 209125
pCMVSport 98 1368 219 795 238 238 237 1 45 46 228 06/19/97 2.0 89
HLHFP03 209126 Uni-ZAP XR 99 613 1 613 224 224 238 1 20 21 116
06/19/97 90 HLNAB07 209126 Lambda ZAP 100 685 1 685 187 187 239 1
32 33 36 06/19/97 II 91 HLWCF05 209126 pCMVSport 101 646 1 646 155
155 240 1 36 37 58 06/19/97 3.0 92 HLYAF80 209126 pSport1 102 826 1
826 222 222 241 1 24 25 47 06/19/97 93 HMDAA66 209126 Uni-ZAP XR
103 586 1 586 106 106 242 1 23 24 31 06/19/97 94 HMKDD07 209126
pSport1 104 628 43 628 267 267 243 1 29 30 63 06/19/97 95 HMKDS08
209126 pSport1 105 558 1 558 230 230 244 1 30 31 67 06/19/97 96
HMSHM14 209126 Uni-ZAP XR 106 756 1 756 103 103 245 1 29 30 45
06/19/97 97 HMWDC28 209126 Uni-Zap XR 107 1146 105 754 124 124 246
1 30 31 42 06/19/97 98 HNDAH54 209126 pCMVSport 108 775 1 775 26 26
247 1 20 21 31 06/19/97 2.0 99 HNFDS53 209126 Uni-ZAP XR 109 911 1
911 200 200 248 1 22 23 23 06/19/97 100 IINFIU96 209126 pBluescript
110 456 1 456 170 170 249 1 33 34 79 06/19/97 101 HNGAC63 209126
Uni-ZAP XR 111 554 1 554 214 214 250 1 15 06/19/97 102 HNGAX58
209126 Uni-ZAP XR 112 722 1 722 100 100 251 1 16 17 46 06/19/97 103
HNGEM24 209126 Uni-ZAP XR 113 931 1 931 239 239 252 1 30 31 31
06/19/97 104 HNGFT78 209126 Uni-ZAP XR 114 588 1 588 20 20 253 1 29
30 35 06/19/97 105 HNHDL85 209126 Uni-ZAP XR 115 812 1 812 194 194
254 1 22 23 50 06/19/97 106 HNHFU59 209126 Uni-ZAP XR 116 506 1 506
278 278 255 1 16 17 76 06/19/97 107 HNHFW22 209126 Uni-ZAP XR 117
751 1 751 228 228 256 1 26 27 60 06/19/97 108 HOAAF80 209126
Uni-ZAP XR 118 960 131 960 303 303 257 1 33 34 36 06/19/97 109
HODCJ90 209126 Uni-ZAP XR 119 1442 326 1133 344 344 258 1 18 19 42
06/19/97 110 HOECO90 209126 Uni-ZAP XR 120 845 215 845 299 299 259
1 24 25 38 06/19/97 111 HPEBT80 209126 Uni-ZAP XR 121 360 1 360 21
21 260 1 40 41 50 06/19/97 112 HSDAG05 209126 Uni-ZAP XR 122 944
231 848 419 419 261 1 37 38 75 06/19/97 113 HSDGR57 209126 Uni-ZAP
XR 123 914 115 914 195 195 262 1 21 22 44 06/19/97 114 HSDJJ82
209126 Uni-ZAP XR 124 462 1 462 79 79 263 1 32 33 52 06/19/97 115
HSDZM95 209126 pBluescript 125 545 1 545 223 223 264 1 23 24 42
06/19/97 116 HSIDI15 209126 Uni-ZAP XR 126 912 1 873 273 273 265 1
22 23 74 06/19/97 117 HSKYU29 209126 pBluescript 127 1048 1 1047
290 290 266 1 36 37 51 06/19/97 118 HSNAA55 209126 Uni-ZAP XR 128
722 1 722 35 35 267 1 15 16 40 06/19/97 119 HSQFP66 209126 Uni-ZAP
XR 129 477 1 477 96 96 268 1 32 33 78 06/19/97 120 HSRDE35 209126
Uni-ZAP XR 130 1296 232 804 428 428 269 1 21 22 116 06/19/97 121
HSSJN64 209126 Uni-ZAP XR 131 738 1 738 70 70 270 1 33 34 61
06/19/97 122 HSVAQ28 209126 Uni-ZAP XR 132 442 1 442 149 149 271 1
24 25 98 06/19/97 123 HSVAY16 209126 Uni-ZAP XR 133 882 1 790 52 52
272 1 30 31 31 06/19/97
[0912] Table 1 summarizes the information corresponding to each
"Gene No." described above. The nucleotide sequence identified as
"NT SEQ ID NO:X" was assembled from partially homologous
("overlapping") sequences obtained from the "cDNA clone ID"
identified in Table 1 and, in some cases, from additional related
DNA clones. The overlapping sequences were assembled into a single
contiguous sequence of high redundancy (usually three to five
overlapping sequences at each nucleotide position), resulting in a
final sequence identified as SEQ ID NO:X.
[0913] The cDNA Clone ID was deposited on the date and given the
corresponding deposit number listed in "ATCC Deposit No:Z and
Date." Some of the deposits contain multiple different clones
corresponding to the same gene. "Vector" refers to the type of
vector contained in the cDNA Clone ID.
[0914] "Total NT Seq." refers to the total number of nucleotides in
the contig identified by "Gene No." The deposited clone may contain
all or most of these sequences, reflected by the nucleotide
position indicated as "5' NT of Clone Seq." and the "3' NT of Clone
Seq." of SEQ ID NO:X. The nucleotide position of SEQ ID NO:X of the
putative start codon (methionine) is identified as "5' NT of Start
Codon." Similarly, the nucleotide position of SEQ ID NO:X of the
predicted signal sequence is identified as "5' NT of First AA of
Signal Pep."
[0915] The translated amino acid sequence, beginning with the
methionine, is identified as "AA SEQ ID NO:Y," although other
reading frames can also be easily translated using known molecular
biology techniques. The polypeptides produced by these alternative
open reading frames are specifically contemplated by the present
invention.
[0916] The first and last amino acid position of SEQ ID NO:Y of the
predicted signal peptide is identified as "First AA of Sig Pep" and
"Last AA of Sig Pep." The predicted first amino acid position of
SEQ ID NO:Y of the secreted portion is identified as "Predicted
First AA of Secreted Portion." Finally, the amino acid position of
SEQ ID NO:Y of the last amino acid in the open reading frame is
identified as "Last AA of ORF."
[0917] SEQ ID NO:X and the translated SEQ ID NO:Y are sufficiently
accurate and otherwise suitable for a variety of uses well known in
the art and described further below. For instance, SEQ ID NO:X is
useful for designing nucleic acid hybridization probes that will
detect nucleic acid sequences contained in SEQ ID NO:X or the cDNA
contained in the deposited clone. These probes will also hybridize
to nucleic acid molecules in biological samples, thereby enabling a
variety of forensic and diagnostic methods of the invention.
Similarly, polypeptides identified from SEQ ID NO:Y may be used to
generate antibodies which bind specifically to the secreted
proteins encoded by the cDNA clones identified in Table 1.
[0918] Nevertheless, DNA sequences generated by sequencing
reactions can contain sequencing errors. The errors exist as
misidentified nucleotides, or as insertions or deletions of
nucleotides in the generated DNA sequence. The erroneously inserted
or deleted nucleotides cause frame shifts in the reading frames of
the predicted amino acid sequence. In these cases, the predicted
amino acid sequence diverges from the actual amino acid sequence,
even though the generated DNA sequence may be greater than 99.9%
identical to the actual DNA sequence (for example, one base
insertion or deletion in an open reading frame of over 1000
bases).
[0919] Accordingly, for those applications requiring precision in
the nucleotide sequence or the amino acid sequence, the present
invention provides not only the generated nucleotide sequence
identified as SEQ ID NO:X and the predicted translated amino acid
sequence identified as SEQ ID NO:Y, but also a sample of plasmid
DNA containing a human cDNA of the invention deposited with the
ATCC, as set forth in Table 1. The nucleotide sequence of each
deposited clone can readily be determined by sequencing the
deposited clone in accordance with known methods. The predicted
amino acid sequence can then be verified from such deposits.
Moreover, the amino acid sequence of the protein encoded by a
particular clone can also be directly determined by peptide
sequencing or by expressing the protein in a suitable host cell
containing the deposited human cDNA, collecting the protein, and
determining its sequence.
[0920] The present invention also relates to the genes
corresponding to SEQ ID NO:X, SEQ ID NO:Y, or the deposited clone.
The corresponding gene can be isolated in accordance with known
methods using the sequence information disclosed herein. Such
methods include preparing probes or primers from the disclosed
sequence and identifying or amplifying the corresponding gene from
appropriate sources of genomic material.
[0921] Also provided in the present invention are species homologs.
Species homologs may be isolated and identified by making suitable
probes or primers from the sequences provided herein and screening
a suitable nucleic acid source for the desired homologue.
[0922] The polypeptides of the invention can be prepared in any
suitable manner. Such polypeptides include isolated naturally
occurring polypeptides, recombinantly produced polypeptides,
synthetically produced polypeptides, or polypeptides produced by a
combination of these methods. Means for preparing such polypeptides
are well understood in the art.
[0923] The polypeptides may be in the form of the secreted protein,
including the mature form. or may be a part of a larger protein,
such as a fusion protein (see below). It is often advantageous to
include an additional amino acid sequence which contains secretory
or leader sequences, pro-sequences, sequences which aid in
purification, such as multiple histidine residues, or an additional
sequence for stability during recombinant production.
[0924] The polypeptides of the present invention are preferably
provided in an isolated form, and preferably are substantially
purified. A recombinantly produced version of a polypeptide,
including the secreted polypeptide, can be substantially purified
by the one-step method described in Smith and Johnson, Gene
67:31-40 (1988). Polypeptides of the invention also can be purified
from natural or recombinant sources using antibodies of the
invention raised against the secreted protein in methods which are
well known in the art.
[0925] Signal Sequences
[0926] Methods for predicting whether a protein has a signal
sequence, as well as the cleavage point for that sequence, are
available. For instance, the method of McGeoch, Virus Res.
3:271-286 (1985), uses the information from a short N-terminal
charged region and a subsequent uncharged region of the complete
(uncleaved) protein. The method of von Heinje, Nucleic Acids Res.
14:4683-4690 (1986) uses the information from the residues
surrounding the cleavage site, typically residues -13 to +2, where
+1 indicates the amino terminus of the secreted protein. The
accuracy of predicting the cleavage points of known mammalian
secretory proteins for each of these methods is in the range of
75-80%. (von Heinje, supra.) However, the two methods do not always
produce the same predicted cleavage point(s) for a given
protein.
[0927] In the present case, the deduced amino acid sequence of the
secreted polypeptide was analyzed by a computer program called
SignalP (Henrik Nielsen et al., Protein Engineering 10: 1-6
(1997)), which predicts the cellular location of a protein based on
the amino acid sequence. As part of this computational prediction
of localization, the methods of McGeoch and von Heinje are
incorporated. The analysis of the amino acid sequences of the
secreted proteins described herein by this program provided the
results shown in Table 1.
[0928] As one of ordinary skill would appreciate, however, cleavage
sites sometimes vary from organism to organism and cannot be
predicted with absolute certainty. Accordingly, the present
invention provides secreted polypeptides having a sequence shown in
SEQ ID NO:Y which have an N-terminus beginning within 5 residues
(i.e., +or -5 residues) of the predicted cleavage point. Similarly,
it is also recognized that in some cases, cleavage of the signal
sequence from a secreted protein is not entirely uniform, resulting
in more than one secreted species. These polypeptides, and the
polynucleotides encoding such polypeptides, are contemplated by the
present invention.
[0929] Moreover, the signal sequence identified by the above
analysis may not necessarily predict the naturally occurring signal
sequence For example, the naturally occurring signal sequence may
be further upstream from the predicted signal sequence. However, it
is likely that the predicted signal sequence will be capable of
directing the secreted protein to the ER. These polypeptides, and
the polynucleotides encoding such polypeptides, are contemplated by
the present invention.
[0930] Polynucleotide and Polypeptide Variants
[0931] "Variant" refers to a polynucleotide or polypeptide
differing from the polynucleotide or polypeptide of the present
invention, but retaining essential properties thereof. Generally,
variants are overall closely similar, and, in many regions,
identical to the polynucleotide or polypeptide of the present
invention.
[0932] By a polynucleotide having a nucleotide sequence at least,
for example, 95% "identical" to a reference nucleotide sequence of
the present invention, it is intended that the nucleotide sequence
of the polynucleotide is identical to the reference sequence except
that the polynucleotide sequence may include up to five point
mutations per each 100 nucleotides of the reference nucleotide
sequence encoding the polypeptide. In other words, to obtain a
polynucleotide having a nucleotide sequence at least 95% identical
to a reference nucleotide sequence, up to 5% of the nucleotides in
the reference sequence may be deleted or substituted with another
nucleotide, or a number of nucleotides up to 5% of the total
nucleotides in the reference sequence may be inserted into the
reference sequence. The query sequence may be an entire sequence
shown in Table 1, the ORF (open reading frame), or any fragement
specified as described herein.
[0933] As a practical matter, whether any particular nucleic acid
molecule or polypeptide is at least 90%, 95%, 96%, 97%, 98% or 99%
identical to a nucleotide sequence of the presence invention can be
determined conventionally using known computer programs. A
preferred method for determing the best overall match between a
query sequence (a sequence of the present invention) and a subject
sequence, also referred to as a global sequence alignment, can be
determined using the FASTDB computer program based on the algorithm
of Brutlag et al. (Comp. App. Biosci. (1990) 6:237-245). In a
sequence alignment the query and subject sequences are both DNA
sequences. An RNA sequence can be compared by converting U's to
T's. The result of said global sequence alignment is in percent
identity. Preferred parameters used in a FASTDB alignment of DNA
sequences to calculate percent identify are: Matrix=Unitary,
k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization
Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty
0.05, Window Size=500 or the lenght of the subject nucleotide
sequence, whichever is shorter.
[0934] If the subject sequence is shorter than the query sequence
because of 5' or 3' deletions, not because of internal deletions, a
manual correction must be made to the results. This is becuase the
FASTDB program does not account for 5' and 3' truncations of the
subject sequence when calculating percent identity. For subject
sequences truncated at the 5' or 3' ends, relative to the the query
sequence, the percent identity is corrected by calculating the
number of bases of the query sequence that are 5' and 3' of the
subject sequence, which are not matched/aligned, as a percent of
the total bases of the query sequence. Whether a nucleotide is
matched/aligned is determined by results of the FASTDB sequence
alignment. This percentage is then subtracted from the percent
identity, calculated by the above FASTDB program using the
specified parameters, to arrive at a final percent identity score.
This corrected score is what is used for the purposes of the
present invention. Only bases outside the 5' and 3' bases of the
subject sequence, as displayed by the FASTDB alignment, which are
not matched/aligned with the query sequence, are calculated for the
purposes of manually adjusting the percent identity score.
[0935] For example, a 90 base subject sequence is aligned to a 100
base query sequence to determine percent identity. The deletions
occur at the 5' end of the subject sequence and therefore, the
FASTDB alignment does not show a matched/alignement of the first 10
bases at 5' end. The 10 unpaired bases represent 10% of the
sequence (number of bases at the 5' and 3' ends not matched/total
number of bases in the query sequence) so 10% is subtracted from
the percent identity score calculated by the FASTDB program. If the
remaining 90 bases were perfectly matched the final percent
identity would be 90%. In another example, a 90 base subject
sequence is compared with a 100 base query sequence. This time the
deletions are internal deletions so that there are no bases on the
5' or 3' of the subject sequence which are not matched/aligned with
the query. In this case the percent identity calculated by FASTDB
is not manually corrected. Once again, only bases 5' and 3' of the
subject sequence which are not matched/aligned with the query
sequnce are manually corrected for. No other manual corrections are
to made for the purposes of the present invention.
[0936] By a polypeptide having an amino acid sequence at least, for
example, 95% "identical" to a query amino acid sequence of the
present invention, it is intended that the amino acid sequence of
the subject polypeptide is identical to the query sequence except
that the subject polypeptide sequence may include up to five amino
acid alterations per each 100 amino acids of the query amino acid
sequence. In other words, to obtain a polypeptide having an amino
acid sequence at least 95% identical to a query amino acid
sequence, up to 5% of the amino acid residues in the subject
sequence may be inserted, deleted, (indels) or substituted with
another amino acid. These alterations of the reference sequence may
occur at the amino or carboxy terminal positions of the reference
amino acid sequence or anywhere between those terminal positions,
interspersed either individually among residues in the reference
sequence or in one or more contiguous groups within the reference
sequence.
[0937] As a practical matter, whether any particular polypeptide is
at least 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance,
the amino acid sequences shown in Table 1 or to the amino acid
sequence encoded by deposited DNA clone can be determined
conventionally using known computer programs. A preferred method
for determing the best overall match between a query sequence (a
sequence of the present invention) and a subject sequence, also
referred to as a global sequence alignment, can be determined using
the FASTDB computer program based on the algorithm of Brutlag et
al. (Comp. App. Biosci. (1990) 6:237-245). In a sequence alignment
the query and subject sequences are either both nucleotide
sequences or both amino acid sequences. The result of said global
sequence alignment is in percent identity. Preferred parameters
used in a FASTDB amino acid alignment are: Matrix=PAM 0, k-tuple=2,
Mismatch Penalty=1, Joining Penalty=20, Randomization Group
Length=0, Cutoff Score=1, Window Size=sequence length, Gap
Penalty=5, Gap Size Penalty=0.05, Window Size=500 or the length of
the subject amino acid sequence, whichever is shorter.
[0938] If the subject sequence is shorter than the query sequence
due to N- or C-terminal deletions, not because of internal
deletions, a manual correction must be made to the results. This is
becuase the FASTDB program does not account for N- and C-terminal
truncations of the subject sequence when calculating global percent
identity. For subject sequences truncated at the N- and C-termini,
relative to the the query sequence, the percent identity is
corrected by calculating the number of residues of the query
sequence that are N- and C-terminal of the subject sequence, which
are not matched/aligned with a corresponding subject residue, as a
percent of the total bases of the query sequence. Whether a residue
is matched/aligned is determined by results of the FASTDB sequence
alignment. This percentage is then subtracted from the percent
identity, calculated by the above FASTDB program using the
specified parameters, to arrive at a final percent identity score.
This final percent identity score is what is used for the purposes
of the present invention. Only residues to the N- and C-termini of
the subject sequence, which are not matched/aligned with the query
sequence, are considered for the purposes of manually adjusting the
percent identity score. That is, only query residue positions
outside the farthest N- and C-terminal residues of the subject
sequence.
[0939] For example, a 90 amino acid residue subject sequence is
aligned with a 100 residue query sequence to determine percent
identity. The deletion occurs at the N-terminus of the subject
sequence and therefore, the FASTDB alignment does not show a
matching/alignment of the first 10 residues at the N-terminus. The
10 unpaired residues represent 10% of the sequence (number of
residues at the N- and C-termini not matched/total number of
residues in the query sequence) so 10% is subtracted from the
percent identity score calculated by the FASTDB program. If the
remaining 90 residues were perfectly matched the final percent
identity would be 90%. In another example, a 90 residue subject
sequence is compared with a 100 residue query sequence. This time
the deletions are internal deletions so there are no residues at
the N- or C-termini of the subject sequence which are not
matched/aligned with the query. In this case the percent identity
calculated by FASTDB is not manually corrected. Once again, only
residue positions outside the N- and C-terminal ends of the subject
sequence, as displayed in the FASTDB alignment, which are not
matched/aligned with the query sequnce are manually corrected for.
No other manual corrections are to made for the purposes of the
present invention.
[0940] The variants may contain alterations in the coding regions,
non-coding regions, or both. Especially preferred are
polynucleotide variants containing alterations which produce silent
substitutions, additions, or deletions, but do not alter the
properties or activities of the encoded polypeptide. Nucleotide
variants produced by silent substitutions due to the degeneracy of
the genetic code are preferred. Moreover, variants in which 5-10,
1-5, or 1-2 amino acids are substituted, deleted, or added in any
combination are also preferred. Polynucleotide variants can be
produced for a variety of reasons, e.g., to optimize codon
expression for a particular host (change codons in the human mRNA
to those preferred by a bacterial host such as E. coli).
[0941] Naturally occurring variants are called "allelic variants,"
and refer to one of several alternate forms of a gene occupying a
given locus on a chromosome of an organism. (Genes II, Lewin, B.,
ed., John Wiley & Sons, New York (1985).) These allelic
variants can vary at either the polynucleotide and/or polypeptide
level. Alternatively, non-naturally occurring variants may be
produced by mutagenesis techniques or by direct synthesis.
[0942] Using known methods of protein engineering and recombinant
DNA technology, variants may be generated to improve or alter the
characteristics of the polypeptides of the present invention. For
instance, one or more amino acids can be deleted from the
N-terminus or C-terminus of the secreted protein without
substantial loss of biological function. The authors of Ron et al.,
J. Biol. Chem. 268: 2984-2988 (1993), reported variant KGF proteins
having heparin binding activity even after deleting 3, 8, or 27
amino-terminal amino acid residues. Similarly, Interferon gamma
exhibited up to ten times higher activity after deleting 8-10 amino
acid residues from the carboxy terminus of this protein. (Dobeli et
al., J. Biotechnology 7:199-216 (1988).)
[0943] Moreover, ample evidence demonstrates that variants often
retain a biological activity similar to that of the naturally
occurring protein. For example, Gayle and coworkers (J. Biol. Chem
268:22105-22111 (1993)) conducted extensive mutational analysis of
human cytokine IL-1a. They used random mutagenesis to generate over
3,500 individual IL-1a mutants that averaged 2.5 amino acid changes
per variant over the entire length of the molecule. Multiple
mutations were examined at every possible amino acid position. The
investigators found that "[m]ost of the molecule could be altered
with little effect on either [binding or biological activity]."
(See, Abstract.) In fact, only 23 unique amino acid sequences, out
of more than 3,500 nucleotide sequences examined, produced a
protein that significantly differed in activity from wild-type.
[0944] Furthermore, even if deleting one or more amino acids from
the N-terminus or C-terminus of a polypeptide results in
modification or loss of one or more biological functions, other
biological activities may still be retained. For example, the
ability of a deletion variant to induce and/or to bind antibodies
which recognize the secreted form will likely be retained when less
than the majority of the residues of the secreted form are removed
from the N-terminus or C-terminus. Whether a particular polypeptide
lacking N- or C-terminal residues of a protein retains such
immunogenic activities can readily be determined by routine methods
described herein and otherwise known in the art.
[0945] Thus, the invention further includes polypeptide variants
which show substantial biological activity. Such variants include
deletions, insertions, inversions, repeats, and substitutions
selected according to general rules known in the art so as have
little effect on activity. For example, guidance concerning how to
make phenotypically silent amino acid substitutions is provided in
Bowie, J. U. et al., Science 247:1306-1310 (1990), wherein the
authors indicate that there are two main strategies for studying
the tolerance of an amino acid sequence to change.
[0946] The first strategy exploits the tolerance of amino acid
substitutions by natural selection during the process of evolution.
By comparing amino acid sequences in different species, conserved
amino acids can be identified. These conserved amino acids are
likely important for protein function. In contrast, the amino acid
positions where substitutions have been tolerated by natural
selection indicates that these positions are not critical for
protein function. Thus, positions tolerating amino acid
substitution could be modified while still maintaining biological
activity of the protein.
[0947] The second strategy uses genetic engineering to introduce
amino acid changes at specific positions of a cloned gene to
identify regions critical for protein function. For example, site
directed mutagenesis or alanine-scanning mutagenesis (introduction
of single alanine mutations at every residue in the molecule) can
be used. (Cunningham and Wells, Science 244:1081-1085 (1989).) The
resulting mutant molecules can then be tested for biological
activity.
[0948] As the authors state, these two strategies have revealed
that proteins are surprisingly tolerant of amino acid
substitutions. The authors further indicate which amino acid
changes are likely to be permissive at certain amino acid positions
in the protein. For example, most buried (within the tertiary
structure of the protein) amino acid residues require nonpolar side
chains, whereas few features of surface side chains are generally
conserved. Moreover, tolerated conservative amino acid
substitutions involve replacement of the aliphatic or hydrophobic
amino acids Ala, Val, Leu and Ile; replacement of the hydroxyl
residues Ser and Thr; replacement of the acidic residues Asp and
Glu; replacement of the amide residues Asn and Gln, replacement of
the basic residues Lys, Arg, and His; replacement of the aromatic
residues Phe, Tyr, and Trp, and replacement of the small-sized
amino acids Ala, Ser, Thr, Met, and Gly.
[0949] Besides conservative amino acid substitution, variants of
the present invention include (i) substitutions with one or more of
the non-conserved amino acid residues, where the substituted amino
acid residues may or may not be one encoded by the genetic code, or
(ii) substitution with one or more of amino acid residues having a
substituent group, or (iii) fusion of the mature polypeptide with
another compound, such as a compound to increase the stability
and/or solubility of the polypeptide (for example, polyethylene
glycol), or (iv) fusion of the polypeptide with additional amino
acids, such as an IgG Fc fusion region peptide, or leader or
secretory sequence, or a sequence facilitating purification. Such
variant polypeptides are deemed to be within the scope of those
skilled in the art from the teachings herein.
[0950] For example, polypeptide variants containing amino acid
substitutions of charged amino acids with other charged or neutral
amino acids may produce proteins with improved characteristics,
such as less aggregation. Aggregation of pharmaceutical
formulations both reduces activity and increases clearance due to
the aggregate's immunogenic activity. (Pinckard et al., Clin. Exp.
Immunol. 2:331-340 (1967); Robbins et al., Diabetes 36: 838-845
(1987); Cleland et al., Crit. Rev. Therapeutic Drug Carrier Systems
10:307-377 (1993).)
[0951] Polynucleotide and Polypeptide Fragments
[0952] In the present invention, a "polynucleotide fragment" refers
to a short polynucleotide having a nucleic acid sequence contained
in the deposited clone or shown in SEQ ID NO:X. The short
nucleotide fragments are preferably at least about 15 nt, and more
preferably at least about 20 nt, still more preferably at least
about 30 nt, and even more preferably, at least about 40 nt in
length. A fragment "at least 20 nt in length," for example, is
intended to include 20 or more contiguous bases from the cDNA
sequence contained in the deposited clone or the nucleotide
sequence shown in SEQ ID NO:X. These nucleotide fragments are
useful as diagnostic probes and primers as discussed herein. Of
course, larger fragments (e.g., 50, 150, 500, 600, 2000
nucleotides) are preferred.
[0953] Moreover, representative examples of polynucleotide
fragments of the invention, include, for example, fragments having
a sequence from about nucleotide number 1-50, 51-100, 101-150,
151-200, 201-250, 251-300, 301-350, 351-400, 401-450, 451-500,
501-550, 551-600, 651-700, 701-750, 751-800, 800-850, 851-900,
901-950, 951-1000, 1001-1050, 1051-1100, 1101-1150, 1151-1200,
1201-1250, 1251-1300, 1301-1350, 1351-1400, 1401-1450, 1451-1500,
1501-1550, 1551-1600, 1601-1650, 1651-1700, 1701-1750, 1751-1800,
1801-1850, 1851-1900, 1901-1950, 1951-2000, or 2001 to the end of
SEQ ID NO:X or the cDNA contained in the deposited clone. In this
context "about" includes the particularly recited ranges, larger or
smaller by several (5, 4, 3, 2, or 1) nucleotides, at either
terminus or at both termini. Preferably, these fragments encode a
polypeptide which has biological activity. More preferably, these
polynucleotides can be used as probes or primers as discussed
herein.
[0954] In the present invention, a "polypeptide fragment" refers to
a short amino acid sequence contained in SEQ ID NO:Y or encoded by
the cDNA contained in the deposited clone. Protein fragments may be
"free-standing," or comprised within a larger polypeptide of which
the fragment forms a part or region, most preferably as a single
continuous region. Representative examples of polypeptide fragments
of the invention, include, for example, fragments from about amino
acid number 1-20, 21-40, 41-60, 61-80, 81-100, 102-120, 121-140,
141-160, or 161 to the end of the coding region. Moreover,
polypeptide fragments can be about 20, 30, 40, 50, 60, 70, 80, 90,
100, 110, 120, 130, 140, or 150 amino acids in length. In this
context "about" includes the particularly recited ranges, larger or
smaller by several (5, 4, 3, 2, or 1) amino acids, at either
extreme or at both extremes.
[0955] Preferred polypeptide fragments include the secreted protein
as well as the mature form. Further preferred polypeptide fragments
include the secreted protein or the mature form having a continuous
series of deleted residues from the amino or the carboxy terminus,
or both. For example, any number of amino acids, ranging from 1-60,
can be deleted from the amino terminus of either the secreted
polypeptide or the mature form. Similarly, any number of amino
acids, ranging from 1-30, can be deleted from the carboxy terminus
of the secreted protein or mature form. Furthermore, any
combination of the above amino and carboxy terminus deletions are
preferred. Similarly, polynucleotide fragments encoding these
polypeptide fragments are also preferred.
[0956] Also preferred are polypeptide and polynucleotide fragments
characterized by structural or functional domains, such as
fragments that comprise alpha-helix and alpha-helix forming
regions, beta-sheet and beta-sheet-forming regions, turn and
turn-forming regions, coil and coil-forming regions, hydrophilic
regions, hydrophobic regions, alpha amphipathic regions, beta
amphipathic regions, flexible regions, surface-forming regions,
substrate binding region, and high antigenic index regions.
Polypeptide fragments of SEQ ID NO:Y falling within conserved
domains are specifically contemplated by the present invention.
Moreover, polynucleotide fragments encoding these domains are also
contemplated.
[0957] Other preferred fragments are biologically active fragments.
Biologically active fragments are those exhibiting activity
similar, but not necessarily identical, to an activity of the
polypeptide of the present invention. The biological activity of
the fragments may include an improved desired activity, or a
decreased undesirable activity.
[0958] Epitopes & Antibodies
[0959] In the present invention, "epitopes" refer to polypeptide
fragments having antigenic or immunogenic activity in an animal,
especially in a human. A preferred embodiment of the present
invention relates to a polypeptide fragment comprising an epitope,
as well as the polynucleotide encoding this fragment. A region of a
protein molecule to which an antibody can bind is defined as an
"antigenic epitope." In contrast, an "immunogenic epitope" is
defined as a part of a protein that elicits an antibody response.
(See, for instance, Geysen et al., Proc. Natl. Acad. Sci. USA
81:3998-4002 (1983).)
[0960] Fragments which function as epitopes may be produced by any
conventional means. (See, e.g., Houghten, R. A., Proc. Natl. Acad.
Sci. USA 82:5131-5135 (1985) further described in U.S. Pat. No.
4,631,211.)
[0961] In the present invention, antigenic epitopes preferably
contain a sequence of at least seven, more preferably at least
nine, and most preferably between about 15 to about 30 amino acids.
Antigenic epitopes are useful to raise antibodies, including
monoclonal antibodies, that specifically bind the epitope. (See,
for instance, Wilson et al., Cell 37:767-778 (1984); Sutcliffe, J.
G. et al., Science 219:660-666 (1983).)
[0962] Similarly, immunogenic epitopes can be used to induce
antibodies according to methods well known in the art. (See, for
instance, Sutcliffe et al., supra; Wilson et al., supra; Chow, M.
et al., Proc. Natl. Acad. Sci. USA 82:910-914; and Bittle, F. J. et
al., J. Gen. Virol. 66:2347-2354 (1985).) A preferred immunogenic
epitope includes the secreted protein. The immunogenic epitopes may
be presented together with a carrier protein, such as an albumin,
to an animal system (such as rabbit or mouse) or, if it is long
enough (at least about 25 amino acids), without a carrier. However,
immunogenic epitopes comprising as few as 8 to 10 amino acids have
been shown to be sufficient to raise antibodies capable of binding
to, at the very least, linear epitopes in a denatured polypeptide
(e.g., in Western blotting.)
[0963] As used herein, the term "antibody" (Ab) or "monoclonal
antibody" (Mab) is meant to include intact molecules as well as
antibody fragments (such as, for example, Fab and F(ab')2
fragments) which are capable of specifically binding to protein.
Fab and F(ab')2 fragments lack the Fc fragment of intact antibody,
clear more rapidly from the circulation, and may have less
non-specific tissue binding than an intact antibody. (Wahl et al.,
J. Nucl. Med. 24:316-325 (1983).) Thus, these fragments are
preferred, as well as the products of a FAB or other immunoglobulin
expression library.
[0964] Moreover, antibodies of the present invention include
chimeric, single chain, and humanized antibodies.
[0965] Fusion Proteins
[0966] Any polypeptide of the present invention can be used to
generate fusion proteins. For example, the polypeptide of the
present invention, when fused to a second protein, can be used as
an antigenic tag. Antibodies raised against the polypeptide of the
present invention can be used to indirectly detect the second
protein by binding to the polypeptide. Moreover, because secreted
proteins target cellular locations based on trafficking signals,
the polypeptides of the present invention can be used as targeting
molecules once fused to other proteins.
[0967] Examples of domains that can be fused to polypeptides of the
present invention include not only heterologous signal sequences,
but also other heterologous functional regions. The fusion does not
necessarily need to be direct, but may occur through linker
sequences.
[0968] Moreover, fusion proteins may also be engineered to improve
characteristics of the polypeptide of the present invention. For
instance, a region of additional amino acids, particularly charged
amino acids, may be added to the N-terminus of the polypeptide to
improve stability and persistence during purification from the host
cell or subsequent handling and storage. Also, peptide moieties may
be added to the polypeptide to facilitate purification. Such
regions may be removed prior to final preparation of the
polypeptide. The addition of peptide moieties to facilitate
handling of polypeptides are familiar and routine techniques in the
art.
[0969] Moreover, polypeptides of the present invention, including
fragments, and specifically epitopes, can be combined with parts of
the constant domain of immunoglobulins (IgG), resulting in chimeric
polypeptides. These fusion proteins facilitate purification and
show an increased half-life in vivo. One reported example describes
chimeric proteins consisting of the first two domains of the human
CD4-polypeptide and various domains of the constant regions of the
heavy or light chains of mammalian immunoglobulins. (EP A 394,827;
Traunecker et al., Nature 331:84-86 (1988).) Fusion proteins having
disulfide-linked dimeric structures (due to the IgG) can also be
more efficient in binding and neutralizing other molecules, than
the monomeric secreted protein or protein fragment alone.
(Fountoulakis et al., J. Biochem. 270:3958-3964 (1995).)
[0970] Similarly, EP-A-O 464 533 (Canadian counterpart 2045869)
discloses fusion proteins comprising various portions of constant
region of immunoglobulin molecules together with another human
protein or part thereof. In many cases, the Fc part in a fusion
protein is beneficial in therapy and diagnosis, and thus can result
in, for example, improved pharmacokinetic properties. (EP-A 0232
262.) Alternatively, deleting the Fc part after the fusion protein
has been expressed, detected, and purified, would be desired. For
example, the Fc portion may hinder therapy and diagnosis if the
fusion protein is used as an antigen for immunizations. In drug
discovery, for example, human proteins, such as hIL-5, have been
fused with Fc portions for the purpose of high-throughput screening
assays to identify antagonists of hIL-5. (See, D. Bennett et al.,
J. Molecular Recognition 8:52-58 (1995); K. Johanson et al., J.
Biol. Chem. 270:9459-9471 (1995).)
[0971] Moreover, the polypeptides of the present invention can be
fused to marker sequences, such as a peptide which facilitates
purification of the fused polypeptide. In preferred embodiments,
the marker amino acid sequence is a hexa-histidine peptide, such as
the tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue,
Chatsworth, Calif., 91311), among others, many of which are
commercially available. As described in Gentz et al., Proc. Natl.
Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine
provides for convenient purification of the fusion protein. Another
peptide tag useful for purification, the "HA" tag, corresponds to
an epitope derived from the influenza hemagglutinin protein.
(Wilson et al., Cell 37:767 (1984).)
[0972] Thus, any of these above fusions can be engineered using the
polynucleotides or the polypeptides of the present invention.
[0973] Vectors, Host Cells, and Protein Production
[0974] The present invention also relates to vectors containing the
polynucleotide of the present invention, host cells, and the
production of polypeptides by recombinant techniques. The vector
may be, for example, a phage, plasmid, viral, or retroviral vector.
Retroviral vectors may be replication competent or replication
defective. In the latter case, viral propagation generally will
occur only in complementing host cells.
[0975] The polynucleotides may be joined to a vector containing a
selectable marker for propagation in a host. Generally, a plasmid
vector is introduced in a precipitate, such as a calcium phosphate
precipitate, or in a complex with a charged lipid. If the vector is
a virus, it may be packaged in vitro using an appropriate packaging
cell line and then transduced into host cells.
[0976] The polynucleotide insert should be operatively linked to an
appropriate promoter, such as the phage lambda PL promoter, the E.
coli lac, trp, phoA and tac promoters, the SV40 early and late
promoters and promoters of retroviral LTRs, to name a few. Other
suitable promoters will be known to the skilled artisan. The
expression constructs will further contain sites for transcription
initiation, termination, and, in the transcribed region, a ribosome
binding site for translation. The coding portion of the transcripts
expressed by the constructs will preferably include a translation
initiating codon at the beginning and a termination codon (UAA, UGA
or UAG) appropriately positioned at the end of the polypeptide to
be translated.
[0977] As indicated, the expression vectors will preferably include
at least one selectable marker. Such markers include dihydrofolate
reductase, G418 or neomycin resistance for eukaryotic cell culture
and tetracycline, kanamycin or ampicillin resistance genes for
culturing in E. coli and other bacteria. Representative examples of
appropriate hosts include, but are not limited to, bacterial cells,
such as E. coli, Streptomyces and Salmonella typhimurium cells;
fungal cells, such as yeast cells; insect cells such as Drosophila
S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS, 293,
and Bowes melanoma cells; and plant cells. Appropriate culture
mediums and conditions for the above-described host cells are known
in the art.
[0978] Among vectors preferred for use in bacteria include pQE70,
pQE60 and pQE-9, available from QIAGEN, Inc.; pBluescript vectors,
Phagescript vectors, pNH8A, pNH16a, pNH18A, pNH46A, available from
Stratagene Cloning Systems, Inc.; and ptrc99a, pKK223-3, pKK233-3,
pDR540, pRIT5 available from Pharmacia Biotech, Inc. Among
preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and
pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL
available from Pharmacia. Other suitable vectors will be readily
apparent to the skilled artisan.
[0979] Introduction of the construct into the host cell can be
effected by calcium phosphate transfection, DEAE-dextran mediated
transfection, cationic lipid-mediated transfection,
electroporation, transduction, infection, or other methods. Such
methods are described in many standard laboratory manuals, such as
Davis et al., Basic Methods In Molecular Biology (1986). It is
specifically contemplated that the polypeptides of the present
invention may in fact be expressed by a host cell lacking a
recombinant vector.
[0980] A polypeptide of this invention can be recovered and
purified from recombinant cell cultures by well-known methods
including ammonium sulfate or ethanol precipitation, acid
extraction, anion or cation exchange chromatography,
phosphocellufose chromatography, hydrophobic interaction
chromatography, affinity chromatography, hydroxylapatite
chromatography and lectin chromatography. Most preferably, high
performance liquid chromatography ("HPLC") is employed for
purification.
[0981] Polypeptides of the present invention, and preferably the
secreted form, can also be recovered from: products purified from
natural sources, including bodily fluids, tissues and cells,
whether directly isolated or cultured; products of chemical
synthetic procedures; and products produced by recombinant
techniques from a prokaryotic or eukaryotic host, including, for
example, bacterial, yeast, higher plant, insect, and mammalian
cells. Depending upon the host employed in a recombinant production
procedure, the polypeptides of the present invention may be
glycosylated or may be non-glycosylated. In addition, polypeptides
of the invention may also include an initial modified methionine
residue, in some cases as a result of host-mediated processes.
Thus, it is well known in the art that the N-terminal methionine
encoded by the translation initiation codon generally is removed
with high efficiency from any protein after translation in all
eukaryotic cells. While the N-terminal methionine on most proteins
also is efficiently removed in most prokaryotes, for some proteins,
this prokaryotic removal process is inefficient, depending on the
nature of the amino acid to which the N-terminal methionine is
covalently linked.
[0982] Uses of the Polynucleotides
[0983] Each of the polynucleotides identified herein can be used in
numerous ways as reagents. The following description should be
considered exemplary and utilizes known techniques.
[0984] The polynucleotides of the present invention are useful for
chromosome identification. There exists an ongoing need to identify
new chromosome markers, since few chromosome marking reagents,
based on actual sequence data (repeat polymorphisms), are presently
available. Each polynucleotide of the present invention can be used
as a chromosome marker.
[0985] Briefly, sequences can be mapped to chromosomes by preparing
PCR primers (preferably 15-25 bp) from the sequences shown in SEQ
ID NO:X. Primers can be selected using computer analysis so that
primers do not span more than one predicted exon in the genomic
DNA. These primers are then used for PCR, screening of somatic cell
hybrids containing individual human chromosomes. Only those hybrids
containing the human gene corresponding to the SEQ ID NO:X will
yield an amplified fragment.
[0986] Similarly, somatic hybrids provide a rapid method of PCR
mapping the polynucleotides to particular chromosomes. Three or
more clones can be assigned per day using a single thermal cycler.
Moreover, sublocalization of the polynucleotides can be achieved
with panels of specific chromosome fragments. Other gene
mapping-strategies that can be used include in situ hybridization,
prescreening with labeled flow-sorted chromosomes, and preselection
by hybridization to construct chromosome specific-cDNA
libraries.
[0987] Precise chromosomal location of the polynucleotides can also
be achieved using fluorescence in situ hybridization (FISH) of a
metaphase chromosomal spread. This technique uses polynucleotides
as short as 500 or 600 bases; however, polynucleotides 2,000-4,000
bp are preferred. For a review of this technique, see Verma et al.,
"Human Chromosomes: a Manual of Basic Techniques," Pergamon Press,
New York (1988).
[0988] For chromosome mapping, the polynucleotides can be used
individually (to mark a single chromosome or a single site on that
chromosome) or in panels (for marking multiple sites and/or
multiple chromosomes). Preferred polynucleotides correspond to the
noncoding regions of the cDNAs because the coding sequences are
more likely conserved within gene families, thus increasing the
chance of cross hybridization during chromosomal mapping.
[0989] Once a polynucleotide has been mapped to a precise
chromosomal location, the physical position of the polynucleotide
can be used in linkage analysis. Linkage analysis establishes
coinheritance between a chromosomal location and presentation of a
particular disease. (Disease mapping data are found, for example,
in V. McKusick, Mendelian Inheritance in Man (available on line
through Johns Hopkins University Welch Medical Library).) Assuming
1 megabase mapping resolution and one gene per 20 kb, a cDNA
precisely localized to a chromosomal region associated with the
disease could be one of 50-500 potential causative genes.
[0990] Thus, once coinheritance is established, differences in the
polynucleotide and the corresponding gene between affected and
unaffected individuals can be examined. First, visible structural
alterations in the chromosomes, such as deletions or
translocations, are examined in chromosome spreads or by PCR. If no
structural alterations exist, the presence of point mutations are
ascertained. Mutations observed in some or all affected
individuals, but not in normal individuals, indicates that the
mutation may cause the disease. However, complete sequencing of the
polypeptide and the corresponding gene from several normal
individuals is required to distinguish the mutation from a
polymorphism. If a new polymorphism is identified, this polymorphic
polypeptide can be used for further linkage analysis.
[0991] Furthermore, increased or decreased expression of the gene
in affected individuals as compared to unaffected individuals can
be assessed using polynucleotides of the present invention. Any of
these alterations (altered expression, chromosomal rearrangement,
or mutation) can be used as a diagnostic or prognostic marker.
[0992] In addition to the foregoing, a polynucleotide can be used
to control gene expression through triple helix formation or
antisense DNA or RNA. Both methods rely on binding of the
polynucleotide to DNA or RNA. For these techniques, preferred
polynucleotides are usually 20 to 40 bases in length and
complementary to either the region of the gene involved in
transcription (triple helix--see Lee et al., Nucl. Acids Res.
6:3073 (1979); Cooney et al., Science 241:456 (1988); and Dervan et
al., Science 251:1360 (1991) ) or to the mRNA itself
(antisense--Okano, J. Neurochem. 56:560 (1991);
Oligodeoxy-nucleotides as Antisense Inhibitors of Gene Expression,
CRC Press, Boca Raton, Fla. (1988).) Triple helix formation
optimally results in a shut-off of RNA transcription from DNA,
while antisense RNA hybridization blocks translation of an mRNA
molecule into polypeptide. Both techniques are effective in model
systems, and the information disclosed herein can be used to design
antisense or triple helix polynucleotides in an effort to treat
disease.
[0993] Polynucleotides of the present invention are also useful in
gene therapy. One goal of gene therapy is to insert a normal gene
into an organism having a defective gene, in an effort to correct
the genetic defect. The polynucleotides disclosed in the present
invention offer a means of targeting such genetic defects in a
highly accurate manner. Another goal is to insert a new gene that
was not present in the host genome, thereby producing a new trait
in the host cell.
[0994] The polynucleotides are also useful for identifying
individuals from minute biological samples. The United States
military, for example, is considering the use of restriction
fragment length polymorphism (RFLP) for identification of its
personnel. In this technique, an individual's genomic DNA is
digested with one or more restriction enzymes, and probed on a
Southern blot to yield unique bands for identifying personnel. This
method does not suffer from the current limitations of "Dog Tags"
which can be lost, switched, or stolen, making positive
identification difficult. The polynucleotides of the present
invention can be used as additional DNA markers for RFLP.
[0995] The polynucleotides of the present invention can also be
used as an alternative to RFLP, by determining the actual
base-by-base DNA sequence of selected portions of an individual's
genome. These sequences can be used to prepare PCR primers for
amplifying and isolating such selected DNA, which can then be
sequenced. Using this technique, individuals can be identified
because each individual will have a unique set of DNA sequences.
Once an unique ID database is established for an individual,
positive identification of that individual, living or dead, can be
made from extremely small tissue samples.
[0996] Forensic biology also benefits from using DNA-based
identification techniques as disclosed herein. DNA sequences taken
from very small biological samples such as tissues, e.g., hair or
skin, or body fluids, e.g., blood, saliva, semen, etc., can be
amplified using PCR. In one prior art technique, gene sequences
amplified from polymorphic loci, such as DQa class II HLA gene, are
used in forensic biology to identify individuals. (Erlich, H., PCR
Technology, Freeman and Co. (1992).) Once these specific
polymorphic loci are amplified, they are digested with one or more
restriction enzymes, yielding an identifying set of bands on a
Southern blot probed with DNA corresponding to the DQa class II HLA
gene. Similarly, polynucleotides of the present invention can be
used as polymorphic markers for forensic purposes.
[0997] There is also a need for reagents capable of identifying the
source of a particular tissue. Such need arises, for example, in
forensics when presented with tissue of unknown origin. Appropriate
reagents can comprise, for example, DNA probes or primers specific
to particular tissue prepared from the sequences of the present
invention. Panels of such reagents can identify tissue by species
and/or by organ type. In a similar fashion, these reagents can be
used to screen tissue cultures for contamination.
[0998] In the very least, the polynucleotides of the present
invention can be used as molecular weight markers on Southern gels,
as diagnostic probes for the presence of a specific mRNA in a
particular cell type, as a probe to "subtract-out" known sequences
in the process of discovering novel polynucleotides, for selecting
and making oligomers for attachment to a "gene chip" or other
support, to raise anti-DNA antibodies using DNA immunization
techniques, and as an antigen to elicit an immune response.
[0999] Uses of the Polypeptides
[1000] Each of the polypeptides identified herein can be used in
numerous ways. The following description should be considered
exemplary and utilizes known techniques.
[1001] A polypeptide of the present invention can be used to assay
protein levels in a biological sample using antibody-based
techniques. For example, protein expression in tissues can be
studied with classical immunohistological methods. (Jalkanen, M.,
et al., J. Cell. Biol. 101:976-985 (1985); Jalkanen, M., et al., J.
Cell . Biol. 105:3087-3096 (1987).) Other antibody-based methods
useful for detecting protein gene expression include immunoassays,
such as the enzyme linked immunosorbent assay (ELISA) and the
radioimmunoassay (RIA). Suitable antibody assay labels are known in
the art and include enzyme labels, such as, glucose oxidase, and
radioisotopes, such as iodine (1251, 1211), carbon (14C), sulfur
(35S), tritium (3H), indium (112In), and technetium (99 mTc), and
fluorescent labels, such as fluorescein and rhodamine, and
biotin.
[1002] In addition to assaying secreted protein levels in a
biological sample, proteins can also be detected in vivo by
imaging. Antibody labels or markers for in vivo imaging of protein
include those detectable by X-radiography, NMR or ESR. For
X-radiography, suitable labels include radioisotopes such as barium
or cesium, which emit detectable radiation but are not overtly
harmful to the subject. Suitable markers for NMR and ESR include
those with a detectable characteristic spin, such as deuterium,
which may be incorporated into the antibody by labeling of
nutrients for the relevant hybridoma.
[1003] A protein-specific antibody or antibody fragment which has
been labeled with an appropriate detectable imaging moiety, such as
a radioisotope (for example, 131I, 112In, 99 mTc), a radio-opaque
substance, or a material detectable by nuclear magnetic resonance,
is introduced (for example, parenterally, subcutaneously, or
intraperitoneally) into the mammal. It will be understood in the
art that the size of the subject and the imaging system used will
determine the quantity of imaging moiety needed to produce
diagnostic images. In the case of a radioisotope moiety, for a
human subject, the quantity of radioactivity injected will normally
range from about 5 to 20 millicuries of 99 mTc. The labeled
antibody or antibody fragment will then preferentially accumulate
at the location of cells which contain the specific protein. In
vivo tumor imaging is described in S. W. Burchiel et al.,
"Immunopharmacokinetics of Radiolabeled Antibodies and Their
Fragments." (Chapter 13 in Tumor Imaging: The Radiochemical
Detection of Cancer, S. W. Burchiel and B. A. Rhodes, eds., Masson
Publishing Inc. (1982).)
[1004] Thus, the invention provides a diagnostic method of a
disorder, which involves (a) assaying the expression of a
polypeptide of the present invention in cells or body fluid of an
individual; (b) comparing the level of gene expression with a
standard gene expression level, whereby an increase or decrease in
the assayed polypeptide gene expression level compared to the
standard expression level is indicative of a disorder.
[1005] Moreover, polypeptides of the present invention can be used
to treat disease. For example, patients can be administered a
polypeptide of the present invention in an effort to replace absent
or decreased levels of the polypeptide (e.g., insulin), to
supplement absent or decreased levels of a different polypeptide
(e.g., hemoglobin S for hemoglobin B), to inhibit the activity of a
polypeptide (e.g., an oncogene), to activate the activity of a
polypeptide (e.g., by binding to a receptor), to reduce the
activity of a membrane bound receptor by competing with it for free
ligand (e.g., soluble TNF receptors used in reducing inflammation),
or to bring about a desired response (e.g., blood vessel
growth).
[1006] Similarly, antibodies directed to a polypeptide of the
present invention can also be used to treat disease. For example,
administration of an antibody directed to a polypeptide of the
present invention can bind and reduce overproduction of the
polypeptide. Similarly, administration of an antibody can activate
the polypeptide, such as by binding to a polypeptide bound to a
membrane (receptor).
[1007] At the very least, the polypeptides of the present invention
can be used as molecular weight markers on SDS-PAGE gels or on
molecular sieve gel filtration columns using methods well known to
those of skill in the art. Polypeptides can also be used to raise
antibodies, which in turn are used to measure protein expression
from a recombinant cell, as a way of assessing transformation of
the host cell. Moreover, the polypeptides of the present invention
can be used to test the following biological activities.
[1008] Biological Activities
[1009] The polynucleotides and polypeptides of the present
invention can be used in assays to test for one or more biological
activities. If these polynucleotides and polypeptides do exhibit
activity in a particular assay, it is likely that these molecules
may be involved in the diseases associated with the biological
activity. Thus, the polynucleotides and polypeptides could be used
to treat the associated disease.
[1010] Immune Activity
[1011] A polypeptide or polynucleotide of the present invention may
be useful in treating deficiencies or disorders of the immune
system, by activating or inhibiting the proliferation,
differentiation, or mobilization (chemotaxis) of immune cells.
Immune cells develop through a process called hematopoiesis,
producing myeloid (platelets, red blood cells, neutrophils, and
macrophages) and lymphoid (B and T lymphocytes) cells from
pluripotent stem cells. The etiology of these immune deficiencies
or disorders may be genetic, somatic, such as cancer or some
autoimmune disorders, acquired (e.g., by chemotherapy or toxins),
or infectious. Moreover, a polynucleotide or polypeptide of the
present invention can be used as a marker or detector of a
particular immune system disease or disorder.
[1012] A polynucleotide or polypeptide of the present invention may
be useful in treating or detecting deficiencies or disorders of
hematopoietic cells. A polypeptide or polynucleotide of the present
invention could be used to increase differentiation and
proliferation of hematopoietic cells, including the
pluripotent.stem cells, in an effort to treat those disorders
associated with a decrease in certain (or many) types hematopoietic
cells. Examples of immunologic deficiency syndromes include, but
are not limited to: blood protein disorders (e.g.
agammaglobulinemia, dysgammaglobulinemia), ataxia telangiectasia,
common variable immunodeficiency, Digeorge Syndrome, HIV infection,
HTLV-BLV infection, leukocyte adhesion deficiency syndrome,
lymphopenia, phagocyte bactericidal dysfunction, severe combined
immunodeficiency (SCIDs), Wiskott-Aldrich Disorder, anemia,
thrombocytopenia, or hemoglobinuria.
[1013] Moreover, a polypeptide or polynucleotide of the present
invention could also be used to modulate hemostatic (the stopping
of bleeding) or thrombolytic activity (clot formation). For
example, by increasing hemostatic or thrombolytic activity, a
polynucleotide or polypeptide of the present invention could be
used to treat blood coagulation disorders (e.g., afibrinogenemia,
factor deficiencies), blood platelet disorders (e.g.
thrombocytopenia), or wounds resulting from trauma, surgery, or
other causes. Alternatively, a polynucleotide or polypeptide of the
present invention that can decrease hemostatic or thrombolytic
activity could be used to inhibit or dissolve clotting. These
molecules could be important in the treatment of heart attacks
(infarction), strokes, or scarring.
[1014] A polynucleotide or polypeptide of the present invention may
also be useful in treating or detecting autoimmune disorders. Many
autoimmune disorders result from inappropriate recognition of self
as foreign material by immune cells. This inappropriate recognition
results in an immune response leading to the destruction of the
host tissue. Therefore, the administration of a polypeptide or
polynucleotide of the present invention that inhibits an immune
response, particularly the proliferation, differentiation, or
chemotaxis of T-cells, may be an effective therapy in preventing
autoimmune disorders.
[1015] Examples of autoimmune disorders that can be treated or
detected by the present invention include, but are not limited to:
Addison's Disease, hemolytic anemia, antiphospholipid syndrome,
rheumatoid arthritis, dermatitis, allergic encephalomyelitis,
glomerulonephritis, Goodpasture's Syndrome, Graves' Disease,
Multiple Sclerosis, Myasthenia Gravis, Neuritis, Ophthalmia,
Bullous Pemphigoid, Pemphigus, Polyendocrinopathies, Purpura,
Reiter's Disease, Stiff-Man Syndrome, Autoimmune Thyroiditis,
Systemic Lupus Erythematosus, Autoimmune Pulmonary Inflammation,
Guillain-Barre Syndrome, insulin dependent diabetes mellitis, and
autoimmune inflammatory eye disease.
[1016] Similarly, allergic reactions and conditions, such as asthma
(particularly allergic asthma) or other respiratory problems, may
also be treated by a polypeptide or polynucleotide of the present
invention. Moreover, these molecules can be used to treat
anaphylaxis, hypersensitivity to an antigenic molecule, or blood
group incompatibility.
[1017] A polynucleotide or polypeptide of the present invention may
also be used to treat and/or prevent organ rejection or
graft-versus-host disease (GVHD). Organ rejection occurs by host
immune cell destruction of the transplanted tissue through an
immune response. Similarly, an immune response is also involved in
GVHD, but, in this case, the foreign transplanted immune cells
destroy the host tissues. The administration of a polypeptide or
polynucleotide of the present invention that inhibits an immune
response, particularly the proliferation, differentiation, or
chemotaxis of T-cells, may be an effective therapy in preventing
organ rejection or GVHD.
[1018] Similarly, a polypeptide, or polynucleotide of the present
invention may also be used to modulate inflammation. For example,
the polypeptide or polynucleotide may inhibit the proliferation and
differentiation of cells involved in an inflammatory response.
These molecules can be used to treat inflammatory conditions, both
chronic and acute conditions, including inflammation associated
with infection (e.g., septic shock, sepsis, or systemic
inflammatory response syndrome (SIRS)), ischemia-reperfusion
injury, endotoxin lethality, arthritis, complement-mediated
hyperacute rejection, nephritis, cytokine or chemokine induced lung
injury, inflammatory bowel disease, Crohn's disease, or resulting
from over production of cytokines (e.g., TNF or IL-1.)
[1019] Hyperproliferative Disorders
[1020] A polypeptide or polynucleotide can be used to treat or
detect hyperproliferative disorders, including neoplasms. A
polypeptide or polynucleotide of the present invention may inhibit
the proliferation of the disorder through direct or indirect
interactions. Alternatively, a polypeptide or polynucleotide of the
present invention may proliferate other cells which can inhibit the
hyperproliferative disorder.
[1021] For example, by increasing an immune response, particularly
increasing antigenic qualities of the hyperproliferative disorder
or by proliferating, differentiating, or mobilizing T-cells,
hyperproliferative disorders can be treated. This immune response
may be increased by either enhancing an existing immune response,
or by initiating a new immune response. Alternatively, decreasing
an immune response may also be a method of treating
hyperproliferative disorders, such as a chemotherapeutic agent.
[1022] Examples of hyperproliferative disorders that can be treated
or detected by a polynucleotide or polypeptide of the present
invention include, but are not limited to neoplasms located in the:
abdomen, bone, breast, digestive system, liver, pancreas,
peritoneum, endocrine glands (adrenal, parathyroid, pituitary,
testicles, ovary, thymus, thyroid), eye, head and neck, nervous
(central and peripheral), lymphatic system, pelvic, skin, soft
tissue, spleen, thoracic, and urogenital.
[1023] Similarly, other hyperproliferative disorders can also be
treated or detected by a polynucleotide or polypeptide of the
present invention. Examples of such hyperproliferative disorders
include, but are not limited to: hypergammaglobulinemia,
lymphoproliferative disorders, paraproteinemias, purpura,
sarcoidosis, Sezary Syndrome, Waldenstron's Macroglobulinemia,
Gaucher's Disease, histiocytosis, and any other hyperproliferative
disease, besides neoplasia, located in an organ system listed
above.
[1024] Infectious Disease
[1025] A polypeptide or polynucleotide of the present invention can
be used to treat or detect infectious agents. For example, by
increasing the immune response, particularly increasing the
proliferation and differentiation of B and/or T cells, infectious
diseases may be treated. The immune response may be increased by
either enhancing an existing immune response, or by initiating a
new immune response. Alternatively, the polypeptide or
polynucleotide of the present invention may also directly inhibit
the infectious agent, without necessarily eliciting an immune
response.
[1026] Viruses are one example of an infectious agent that can
cause disease or symptoms that can be treated or detected by a
polynucleotide or polypeptide of the present invention. Examples of
viruses, include, but are not limited to the following DNA and RNA
viral families: Arbovirus, Adenoviridae, Arenaviridae, Arterivirus,
Birnaviridae, Bunyaviridae, Caliciviridae, Circoviridae,
Coronaviridae, Flaviviridae, Hepadnaviridae (Hepatitis),
Herpesviridae (such as, Cytomegalovirus, Herpes Simplex, Herpes
Zoster), Mononegavirus (e.g., Paramyxoviridae, Morbillivirus,
Rhabdoviridae), Orthomyxoviridae (e.g., Influenza), Papovaviridae,
Parvoviridae, Picomaviridae, Poxviridae (such as Smallpox or
Vaccinia), Reoviridae (e.g., Rotavirus), Retroviridae (HTLV-I,
HTLV-II, Lentivirus), and Togaviridae (e.g., Rubivirus). Viruses
falling within these families can cause a variety of diseases or
symptoms, including, but not limited to: arthritis, bronchiollitis,
encephalitis, eye infections (e.g., conjunctivitis, keratitis),
chronic fatigue syndrome, hepatitis (A, B, C, E, Chronic Active,
Delta), meningitis, opportunistic infections (e.g., AIDS),
pneumonia, Burkitt's Lymphoma, chickenpox, hemorrhagic fever,
Measles, Mumps, Parainfluenza, Rabies, the common cold, Polio,
leukemia, Rubella, sexually transmitted-diseases, skin diseases
(e.g.; Kaposi's, warts), and viremia. A polypeptide or
polynucleotide of the present invention can be used to treat or
detect any of these symptoms or diseases.
[1027] Similarly, bacterial or fungal agents that can cause disease
or symptoms and that can be treated or detected by a polynucleotide
or polypeptide of the present invention include, but not limited
to, the following Gram-Negative and Gram-positive bacterial
families and fungi: Actinomycetales (e.g., Corynebacterium,
Mycobacterium, Norcardia), Aspergillosis, Bacillaceae (e.g.,
Anthrax, Clostridium), Bacteroidaceae, Blastomycosis, Bordetella,
Borrelia, Brucellosis, Candidiasis, Campylobacter,
Coccidioidomycosis, Cryptococcosis, Dermatocycoses,
Enterobacteriaceae (Kiebsiella, Salmonella, Serratia, Yersinia),
Erysipelothrix, Helicobacter, Legionellosis, Leptospirosis,
Listeria, Mycoplasmatales, Neisseriaceae (e.g., Acinetobacter,
Gonorrhea, Menigococcal), Pasteurellacea Infections (e.g.,
Actinobacillus, Heamophilus, Pasteurella), Pseudomonas,
Rickettsiaceae. Chlamydiaceae, Syphilis, and Staphylococcal. These
bacterial or fungal families can cause the following diseases or
symptoms, including, but not limited to: bacteremia, endocarditis,
eye infections (conjunctivitis, tuberculosis, uveitis), gingivitis,
opportunistic infections (e.g., AIDS related infections),
paronychia, prosthesis-related infections, Reiter's Disease,
respiratory tract infections, such as Whooping Cough or Empyema,
sepsis, Lyme Disease, Cat-Scratch Disease, Dysentery, Paratyphoid
Fever, food poisoning, Typhoid, pneumonia, Gonorrhea, meningitis,
Chlamydia, Syphilis, Diphtheria, Leprosy, Paratuberculosis,
Tuberculosis, Lupus, Botulism, gangrene, tetanus, impetigo,
Rheumatic Fever, Scarlet Fever, sexually transmitted diseases, skin
diseases (e.g., cellulitis, dermatocycoses), toxemia, urinary tract
infections, wound infections. A polypeptide or polynucleotide of
the present invention can be used to treat or detect any of these
symptoms or diseases.
[1028] Moreover, parasitic agents causing disease or symptoms that
can be treated or detected by a polynucleotide or polypeptide of
the present invention include, but not limited to, the following
families: Amebiasis, Babesiosis, Coccidiosis, Cryptosporidiosis,
Dientamoebiasis, Dourine, Ectoparasitic, Giardiasis, Helminthiasis,
Leishmaniasis, Theileriasis, Toxoplasmosis, Trypanosomiasis, and
Trichomonas. These parasites can cause a variety of diseases or
symptoms, including, but not limited to: Scabies, Trombiculiasis,
eye infections, intestinal disease (e.g., dysentery, giardiasis),
liver disease, lung disease, opportunistic infections (e.g., AIDS
related), Malaria, pregnancy complications, and toxoplasmosis. A
polypeptide or polynucleotide of the present invention can be used
to treat or detect any of these symptoms or diseases.
[1029] Preferably, treatment using a polypeptide or polynucleotide
of the present invention could either be by administering an
effective amount of a polypeptide to the patient, or by removing
cells from the patient, supplying the cells with a polynucleotide
of the present invention, and returning the engineered cells to the
patient (ex vivo therapy). Moreover, the polypeptide or
polynucleotide of the present invention can be used as an antigen
in a vaccine to raise an immune response against infectious
disease.
[1030] Regeneration
[1031] A polynucleotide or polypeptide of the present invention can
be used to differentiate, proliferate, and attract cells, leading
to the regeneration of tissues. (See, Science 276:59-87 (1997).)
The regeneration of tissues could be used to repair, replace, or
protect tissue damaged by congenital defects, trauma (wounds,
burns, incisions, or ulcers), age, disease (e.g. osteoporosis,
osteocarthritis, periodontal disease, liver failure), surgery,
including cosmetic plastic surgery, fibrosis, reperfusion injury,
or systemic cytokine damage.
[1032] Tissues that could be regenerated using the present
invention include organs (e.g., pancreas, liver, intestine, kidney,
skin, endothelium), muscle (smooth, skeletal or cardiac), vascular
(including vascular endothelium), nervous, hematopoietic, and
skeletal (bone, cartilage, tendon, and ligament) tissue.
Preferably, regeneration occurs without or decreased scarring.
Regeneration also may include angiogenesis.
[1033] Moreover, a polynucleotide or polypeptide of the present
invention may increase regeneration of tissues difficult to heal.
For example, increased tendon/ligament regeneration would quicken
recovery time after damage. A polynucleotide or polypeptide of the
present invention could also be used prophylactically in an effort
to avoid damage. Specific diseases that could be treated include of
tendinitis, carpal tunnel syndrome, and other tendon or ligament
defects. A further example of tissue regeneration of non-healing
wounds includes pressure ulcers, ulcers associated with vascular
insufficiency, surgical, and traumatic wounds.
[1034] Similarly, nerve and brain tissue could also be regenerated
by using a polynucleotide or polypeptide of the present invention
to proliferate and differentiate nerve cells. Diseases that could
be treated using this method include central and peripheral nervous
system diseases, neuropathies, or mechanical and traumatic
disorders (e.g., spinal cord disorders, head trauma,
cerebrovascular disease, and stoke). Specifically, diseases
associated with peripheral nerve injuries, peripheral neuropathy
(e.g., resulting from chemotherapy or other medical therapies),
localized neuropathies, and central nervous system diseases (e.g.,
Alzheimer's disease, Parkinson's disease, Huntington's disease,
amyotrophic lateral sclerosis, and Shy-Drager syndrome), could all
be treated using the polynucleotide or polypeptide of the present
invention.
[1035] Chemotaxis
[1036] A polynucleotide or polypeptide of the present invention may
have chemotaxis activity. A chemotaxic molecule attracts or
mobilizes cells (e.g., monocytes, fibroblasts, neutrophils,
T-cells, mast cells, eosinophils, epithelial and/or endothelial
cells) to a particular site in the body, such as inflammation,
infection, or site of hyperproliferation. The mobilized cells can
then fight off and/or heal the particular trauma or
abnormality.
[1037] A polynucleotide or polypeptide of the present invention may
increase chemotaxic activity of particular cells. These chemotactic
molecules can then be used to treat inflammation, infection,
hyperproliferative disorders, or any immune system disorder by
increasing the number of cells targeted to a particular location in
the body. For example, chemotaxic molecules can be used to treat
wounds and other trauma to tissues by attracting immune cells to
the injured location. Chemotactic molecules of the present
invention can also attract fibroblasts, which can be used to treat
wounds.
[1038] It is also contemplated that a polynucleotide or polypeptide
of the present invention may inhibit chemotactic activity. These
molecules could also be used to treat disorders. Thus, a
polynucleotide or polypeptide of the present invention could be
used as an inhibitor of chemotaxis.
[1039] Binding Activity
[1040] A polypeptide of the present invention may be used to screen
for molecules that bind to the polypeptide or for molecules to
which the polypeptide binds. The binding of the polypeptide and the
molecule may activate (agonist), increase, inhibit (antagonist), or
decrease activity of the polypeptide or the molecule bound.
Examples of such molecules include antibodies, oligonucleotides,
proteins (e.g., receptors),or small molecules.
[1041] Preferably, the molecule is closely related to the natural
ligand of the polypeptide, e.g., a fragment of the ligand, or a
natural substrate, a ligand, a structural or functional mimetic.
(See, Coligan et al., Current Protocols in Immunology 1(2):Chapter
5 (1991).) Similarly, the molecule can be closely related to the
natural receptor to which the polypeptide binds, or at least, a
fragment of the receptor capable of being bound by the polypeptide
(e.g., active site). In either case, the molecule can be rationally
designed using known techniques.
[1042] Preferably, the screening for these molecules involves
producing appropriate cells which express the polypeptide, either
as a secreted protein or on the cell membrane. Preferred cells
include cells from mammals, yeast, Drosophila, or E. coli. Cells
expressing the polypeptide (or cell membrane containing the
expressed polypeptide) are then preferably contacted with a test
compound potentially containing the molecule to observe binding,
stimulation, or inhibition of activity of either the polypeptide or
the molecule.
[1043] The assay may simply test binding of a candidate compound to
the polypeptide, wherein binding is detected by a label, or in an
assay involving competition with a labeled competitor. Further, the
assay may test whether the candidate compound results in a signal
generated by binding to the polypeptide.
[1044] Alternatively, the assay can be carried out using cell-free
preparations, polypeptide/molecule affixed to a solid support,
chemical libraries, or natural product mixtures. The assay may also
simply comprise the steps of mixing a candidate compound with a
solution containing a polypeptide, measuring polypeptide/molecule
activity or binding, and comparing the polypeptide/molecule
activity or binding to a standard.
[1045] Preferably, an ELISA assay can measure polypeptide level or
activity in a sample (e.g., biological sample) using a monoclonal
or polyclonal antibody. The antibody can measure polypeptide level
or activity by either binding, directly or indirectly, to the
polypeptide or by competing with the polypeptide for a
substrate.
[1046] All of these above assays can be used as diagnostic or
prognostic markers. The molecules discovered using these assays can
be used to treat disease or to bring about a particular result in a
patient (e.g., blood vessel growth) by activating or inhibiting the
polypeptide/molecule. Moreover, the assays can discover agents
which may inhibit or enhance the production of the polypeptide from
suitably manipulated cells or tissues.
[1047] Therefore, the invention includes a method of identifying
compounds which bind to a polypeptide of the invention comprising
the steps of: (a) incubating a candidate binding compound with a
polypeptide of the invention; and (b) determining if binding has
occurred. Moreover, the invention includes a method of identifying
agonists/antagonists comprising the steps of: (a) incubating a
candidate compound with a polypeptide of the invention, (b)
assaying a biological activity, and (b) determining if a biological
activity of the polypeptide has been altered.
[1048] Other Activities
[1049] A polypeptide or polynucleotide of the present invention may
also increase or decrease the differentiation or proliferation of
embryonic stem cells, besides, as discussed above, hematopoietic
lineage.
[1050] A polypeptide or polynucleotide of the present invention may
also be used to modulate mammalian characteristics, such as body
height, weight, hair color, eye color, skin, percentage of adipose
tissue, pigmentation, size, and shape (e.g., cosmetic surgery).
Similarly, a polypeptide or polynucleotide of the present invention
may be used to modulate mammalian metabolism affecting catabolism,
anabolism, processing, utilization, and storage of energy.
[1051] A polypeptide or polynucleotide of the present invention may
be used to change a mammal's mental state or physical state by
influencing biorhythms, caricadic rhythms, depression (including
depressive disorders), tendency for violence, tolerance for pain,
reproductive capabilities (preferably by Activin or Inhibin-like
activity), hormonal or endocrine levels, appetite, libido, memory,
stress, or other cognitive qualities.
[1052] A polypeptide or polynucleotide of the present invention may
also be used as a food additive or preservative, such as to
increase or decrease storage capabilities, fat content, lipid,
protein, carbohydrate, vitamins, minerals, cofactors or other
nutritional components.
[1053] Other Preferred Embodiments
[1054] Other preferred embodiments of the claimed invention include
an isolated nucleic acid molecule comprising a nucleotide sequence
which is at least 95% identical to a sequence of at least about 50
contiguous nucleotides in the nucleotide sequence of SEQ ID NO:X
wherein X is any integer as defined in Table 1.
[1055] Also preferred is a nucleic acid molecule wherein said
sequence of contiguous nucleotides is included in the nucleotide
sequence of SEQ ID NO:X in the range of positions beginning with
the nucleotide at about the position of the 5' Nucleotide of the
Clone Sequence and ending with the nucleotide at about the position
of the 3' Nucleotide of the Clone Sequence as defined for SEQ ID
NO:X in Table 1.
[1056] Also preferred is a nucleic acid molecule wherein said
sequence of contiguous nucleotides is included in the nucleotide
sequence of SEQ ID NO:X in the range of positions beginning with
the nucleotide at about the position of the 5' Nucleotide of the
Start Codon and ending with the nucleotide at about the position of
the 3' Nucleotide of the Clone Sequence as defined for SEQ ID NO:X
in Table 1.
[1057] Similarly preferred is a nucleic acid molecule wherein said
sequence of contiguous nucleotides is included in the nucleotide
sequence of SEQ ID NO:X in the range of positions beginning with
the nucleotide at about the position of the 5' Nucleotide of the
First Amino Acid of the Signal Peptide and ending with the
nucleotide at about the position of the 3' Nucleotide of the Clone
Sequence as defined for SEQ ID NO:X in Table 1.
[1058] Also preferred is an isolated nucleic acid molecule
comprising a nucleotide sequence which is at least 95% identical to
a sequence of at least about 150 contiguous nucleotides in the
nucleotide sequence of SEQ ID NO:X.
[1059] Further preferred is an isolated nucleic acid molecule
comprising a nucleotide sequence which is at least 95% identical to
a sequence of at least about 500 contiguous nucleotides in the
nucleotide sequence of SEQ ID NO:X.
[1060] A further preferred embodiment is a nucleic acid molecule
comprising a nucleotide sequence which is at least 95% identical to
the nucleotide sequence of SEQ ID NO:X beginning with the
nucleotide at about the position of the 5' Nucleotide of the First
Amino Acid of the Signal Peptide and ending with the nucleotide at
about the position of the 3' Nucleotide of the Clone Sequence as
defined for SEQ ID NO:X in Table 1.
[1061] A further preferred embodiment is an isolated nucleic acid
molecule comprising a nucleotide sequence which is at least 95%
identical to the complete nucleotide sequence of SEQ ID NO:X.
[1062] Also preferred is an isolated nucleic acid molecule which
hybridizes under stringent hybridization conditions to a nucleic
acid molecule, wherein said nucleic acid molecule which hybridizes
does not hybridize under stringent hybridization conditions to a
nucleic acid molecule having a nucleotide sequence consisting of
only A residues or of only T residues.
[1063] Also preferred is a composition of matter comprising a DNA
molecule which comprises a human cDNA clone identified by a cDNA
Clone Identifier in Table 1, which DNA molecule is contained in the
material deposited with the American Type Culture Collection and
given the ATCC Deposit Number shown in Table 1 for said cDNA Clone
Identifier.
[1064] Also preferred is an isolated nucleic acid molecule
comprising a nucleotide sequence which is at least 95% identical to
a sequence of at least 50 contiguous nucleotides in the nucleotide
sequence of a human cDNA clone identified by a cDNA Clone
Identifier in Table 1, which DNA molecule is contained in the
deposit given the ATCC Deposit Number shown in Table 1.
[1065] Also preferred is an isolated nucleic acid molecule, wherein
said sequence of at least 50 contiguous nucleotides is included in
the nucleotide sequence of the complete open reading frame sequence
encoded by said human cDNA clone.
[1066] Also preferred is an isolated nucleic acid molecule
comprising a nucleotide sequence which is at least 95% identical to
sequence of at least 150 contiguous nucleotides in the nucleotide
sequence encoded by said human cDNA clone.
[1067] A further preferred embodiment is an isolated nucleic acid
molecule comprising a nucleotide sequence which is at least 95%
identical to sequence of at least 500 contiguous nucleotides in the
nucleotide sequence encoded by said human cDNA clone.
[1068] A further preferred embodiment is an isolated nucleic acid
molecule comprising a nucleotide sequence which is at least 95%
identical to the complete nucleotide sequence encoded by said human
cDNA clone.
[1069] A further preferred embodiment is a method for detecting in
a biological sample a nucleic acid molecule comprising a nucleotide
sequence which is at least 95% identical to a sequence of at least
50 contiguous nucleotides in a sequence selected from the group
consisting of: a nucleotide sequence of SEQ ID NO:X wherein X is
any integer as defined in Table 1; and a nucleotide sequence
encoded by a human cDNA clone identified by a cDNA Clone Identifier
in Table 1 and contained in the deposit with the ATCC Deposit
Number shown for said cDNA clone in Table 1; which method comprises
a step of comparing a nucleotide sequence of at least one nucleic
acid molecule in said sample with a sequence selected from said
group and determining whether the sequence of said nucleic acid
molecule in said sample is at least 95% identical to said selected
sequence.
[1070] Also preferred is the above method wherein said step of
comparing sequences comprises determining the extent of nucleic
acid hybridization between nucleic acid molecules in said sample
and a nucleic acid molecule comprising said sequence selected from
said group. Similarly, also preferred is the above method wherein
said step of comparing sequences is performed by comparing the
nucleotide sequence determined from a nucleic acid molecule in said
sample with said sequence selected from said group. The nucleic
acid molecules can comprise DNA molecules or RNA molecules.
[1071] A further preferred embodiment is a method for identifying
the species, tissue or cell type of a biological sample which
method comprises a step of detecting nucleic acid molecules in said
sample, if any, comprising a nucleotide sequence that is at least
95% identical to a sequence of at least 50 contiguous nucleotides
in a sequence selected from the group consisting of: a nucleotide
sequence of SEQ ID NO:X wherein X is any integer as defined in
Table 1; and a nucleotide sequence encoded by a human cDNA clone
identified by a cDNA Clone Identifier in Table 1 and contained in
the deposit with the ATCC Deposit Number shown for said cDNA clone
in Table 1.
[1072] The method for identifying the species, tissue or cell type
of a biological sample can comprise a step of detecting nucleic
acid molecules comprising a nucleotide sequence in a panel of at
least two nucleotide sequences, wherein at least one sequence in
said panel is at least 95% identical to a sequence of at least 50
contiguous nucleotides in a sequence selected from said group.
[1073] Also preferred is a method for diagnosing in a subject a
pathological condition associated with abnormal structure or
expression of a gene encoding a secreted protein identified in
Table 1, which method comprises a step of detecting in a biological
sample obtained from said subject nucleic acid molecules, if any,
comprising a nucleotide sequence that is at least 95% identical to
a sequence of at least 50 contiguous nucleotides in a sequence
selected from the group consisting of: a nucleotide sequence of SEQ
ID NO:X wherein X is any integer as defined in Table 1; and a
nucleotide sequence encoded by a human cDNA clone identified by a
cDNA Clone Identifier in Table 1 and contained in the deposit with
the ATCC Deposit Number shown for said cDNA clone in Table 1.
[1074] The method for diagnosing a pathological condition can
comprise a step of detecting nucleic acid molecules comprising a
nucleotide sequence in a panel of at least two nucleotide
sequences, wherein at least one sequence in said panel is at least
95% identical to a sequence of at least 50 contiguous nucleotides
in a sequence selected from said group.
[1075] Also preferred is a composition of matter comprising
isolated nucleic acid molecules wherein the nucleotide sequences of
said nucleic acid molecules comprise a panel of at least two
nucleotide sequences, wherein at least one sequence in said panel
is at least 95% identical to a sequence of at least 50 contiguous
nucleotides in a sequence selected from the group consisting of: a
nucleotide sequence of SEQ ID NO:X wherein X is any integer as
defined in Table 1; and a nucleotide sequence encoded by a human
cDNA clone identified by a cDNA Clone Identifier in Table 1 and
contained in the deposit with the ATCC Deposit Number shown for
said cDNA clone in Table 1. The nucleic acid molecules can comprise
DNA molecules or RNA molecules.
[1076] Also preferred is an isolated polypeptide comprising an
amino acid sequence at least 90% identical to a sequence of at
least about 10 contiguous amino acids in the amino acid sequence of
SEQ ID NO:Y wherein Y is any integer as defined in Table 1.
[1077] Also preferred is a polypeptide, wherein said sequence of
contiguous amino acids is included in the amino acid sequence of
SEQ ID NO:Y in the range of positions beginning with the residue at
about the position of the First Amino Acid of the Secreted Portion
and ending with the residue at about the Last Amino Acid of the
Open Reading Frame as set forth for SEQ ID NO:Y in Table 1.
[1078] Also preferred is an isolated polypeptide comprising an
amino acid sequence at least 95% identical to a sequence of at
least about 30 contiguous amino acids in the amino acid sequence of
SEQ ID NO:Y.
[1079] Further preferred is an isolated polypeptide comprising an
amino acid sequence at least 95% identical to a sequence of at
least about 100 contiguous amino acids in the amino acid sequence
of SEQ ID NO:Y.
[1080] Further preferred is an isolated polypeptide comprising an
amino acid sequence at least 95% identical to the complete amino
acid sequence of SEQ ID NO:Y.
[1081] Further preferred is an isolated polypeptide comprising an
amino acid sequence at least 90% identical to a sequence of at
least about 10 contiguous amino acids in the complete amino acid
sequence of a secreted protein encoded by a human cDNA clone
identified by a cDNA Clone Identifier in Table 1 and contained in
the deposit with the ATCC Deposit Number shown for said cDNA clone
in Table 1.
[1082] Also preferred is a polypeptide wherein said sequence of
contiguous amino acids is included in the amino acid sequence of a
secreted portion of the secreted protein encoded by a human cDNA
clone identified by a cDNA Clone Identifier in Table 1 and
contained in the deposit with the ATCC Deposit Number shown for
said cDNA clone in Table 1.
[1083] Also preferred is an isolated polypeptide comprising an
amino acid sequence at least 95% identical to a sequence of at
least about 30 contiguous amino acids in the amino acid sequence of
the secreted portion of the protein encoded by a human cDNA clone
identified by a cDNA Clone Identifier in Table 1 and contained in
the deposit with the ATCC Deposit Number shown for said cDNA clone
in Table 1.
[1084] Also preferred is an isolated polypeptide comprising an
amino acid sequence at least 95% identical to a sequence of at
least about 100 contiguous amino acids in the amino acid sequence
of the secreted portion of the protein encoded by a human cDNA
clone identified by a cDNA Clone Identifier in Table 1 and
contained in the deposit with the ATCC Deposit Number shown for
said cDNA clone in Table 1.
[1085] Also preferred is an isolated polypeptide comprising an
amino acid sequence at least 95% identical to the amino acid
sequence of the secreted portion of the protein encoded by a human
cDNA clone identified by a cDNA Clone Identifier in Table 1 and
contained in the deposit with the ATCC Deposit Number shown for
said cDNA clone in Table 1.
[1086] Further preferred is an isolated antibody which binds
specifically to a polypeptide comprising an amino acid sequence
that is at least 90% identical to a sequence of at least 10
contiguous amino acids in a sequence selected from the group
consisting of: an amino acid sequence of SEQ ID NO:Y wherein Y is
any integer as defined in Table 1; and a complete amino acid
sequence of a protein encoded by a human cDNA clone identified by a
cDNA Clone Identifier in Table 1 and contained in the deposit with
the ATCC Deposit Number shown for said cDNA clone in Table 1.
[1087] Further preferred is a method for detecting in a biological
sample a polypeptide comprising an amino acid sequence which is at
least 90% identical to a sequence of at least 10 contiguous amino
acids in a sequence selected from the group consisting of: an amino
acid sequence of SEQ ID NO:Y wherein Y is any integer as defined in
Table 1; and a complete amino acid sequence of a protein encoded by
a human cDNA clone identified by a cDNA Clone Identifier in Table 1
and contained in the deposit with the ATCC Deposit Number shown for
said cDNA clone in Table 1; which method comprises a step of
comparing an amino acid sequence of at least one polypeptide
molecule in said sample with a sequence selected from said group
and determining whether the sequence of said polypeptide molecule
in said sample is at least 90% identical to said sequence of at
least 10 contiguous amino acids.
[1088] Also preferred is the above method wherein said step of
comparing an amino acid sequence of at least one polypeptide
molecule in said sample with a sequence selected from said group
comprises determining the extent of specific binding of
polypeptides in said sample to an antibody which binds specifically
to a polypeptide comprising an amino acid sequence that is at least
90% identical to a sequence of at least 10 contiguous amino acids
in a sequence selected from the group consisting of: an amino acid
sequence of SEQ ID NO:Y wherein Y is any integer as defined in
Table 1; and a complete amino acid sequence of a protein encoded by
a human cDNA clone identified by a cDNA Clone Identifier in Table 1
and contained in the deposit with the ATCC Deposit Number shown for
said cDNA clone in Table 1.
[1089] Also preferred is the above method wherein said step of
comparing sequences is performed by comparing the amino acid
sequence determined from a polypeptide molecule in said sample with
said sequence selected from said group.
[1090] Also preferred is a method for identifying the species,
tissue or cell type of a biological sample which method comprises a
step of detecting polypeptide molecules in said sample, if any,
comprising an amino acid sequence that is at least 90% identical to
a sequence of at least 10 contiguous amino acids in a sequence
selected from the group consisting of: an amino acid sequence of
SEQ ID NO:Y wherein Y is any integer as defined in Table 1; and a
complete amino acid sequence of a secreted protein encoded by a
human cDNA clone identified by a cDNA Clone Identifier in Table 1
and contained in the deposit with the ATCC Deposit Number shown for
said cDNA clone in Table 1.
[1091] Also preferred is the above method for identifying the
species, tissue or cell type of a biological sample, which method
comprises a step of detecting polypeptide molecules comprising an
amino acid sequence in a panel of at least two amino acid
sequences, wherein at least one sequence in said panel is at least
90% identical to a sequence of at least 10 contiguous amino acids
in a sequence selected from the above group.
[1092] Also preferred is a method for diagnosing in a subject a
pathological condition associated with abnormal structure or
expression of a gene encoding a secreted protein identified in
Table 1, which method comprises a step of detecting in a biological
sample obtained from said subject polypeptide molecules comprising
an amino acid sequence in a panel of at least two amino acid
sequences, wherein at least one sequence in said panel is at least
90% identical to a sequence of at least 10 contiguous amino acids
in a sequence selected from the group consisting of: an amino acid
sequence of SEQ ID NO:Y wherein Y is any integer as defined in
Table 1; and a complete amino acid sequence of a secreted protein
encoded by a human cDNA clone identified by a cDNA Clone Identifier
in Table 1 and contained in the deposit with the ATCC Deposit
Number shown for said cDNA clone in Table 1.
[1093] In any of these methods, the step of detecting said
polypeptide molecules includes using an antibody.
[1094] Also preferred is an isolated nucleic acid molecule
comprising a nucleotide sequence which is at least 95% identical to
a nucleotide sequence encoding a polypeptide wherein said
polypeptide comprises an amino acid sequence that is at least 90%
identical to a sequence of at least 10 contiguous amino acids in a
sequence selected from the group consisting of: an amino acid
sequence of SEQ ID NO:Y wherein Y is any integer as defined in
Table 1; and a complete amino acid sequence of a secreted protein
encoded by a human cDNA clone identified by a cDNA Clone Identifier
in Table 1 and contained in the deposit with the ATCC Deposit
Number shown for said cDNA clone in Table 1.
[1095] Also preferred is an isolated nucleic acid molecule, wherein
said nucleotide sequence encoding a polypeptide has been optimized
for expression of said polypeptide in a prokaryotic host.
[1096] Also preferred is an isolated nucleic acid molecule, wherein
said polypeptide comprises an amino acid sequence selected from the
group consisting of: an amino acid sequence of SEQ ID NO:Y wherein
Y is any integer as defined in Table 1; and a complete amino acid
sequence of a secreted protein encoded by a human cDNA clone
identified by a cDNA Clone Identifier in Table 1 and contained in
the deposit with the ATCC Deposit Number shown for said cDNA clone
in Table 1.
[1097] Further preferred is a method of making a recombinant vector
comprising inserting any of the above isolated nucleic acid
molecule into a vector. Also preferred is the recombinant vector
produced by this method. Also preferred is a method of making a
recombinant host cell comprising introducing the vector into a host
cell, as well as the recombinant host cell produced by this
method.
[1098] Also preferred is a method of making an isolated polypeptide
comprising culturing this recombinant host cell under conditions
such that said polypeptide is expressed and recovering said
polypeptide. Also preferred is this method of making an isolated
polypeptide, wherein said recombinant host cell is a eukaryotic
cell and said polypeptide is a secreted portion of a human secreted
protein comprising an amino acid sequence selected from the group
consisting of: an amino acid sequence of SEQ ID NO:Y beginning with
the residue at the position of the First Amino Acid of the Secreted
Portion of SEQ ID NO:Y wherein Y is an integer set forth in Table 1
and said position of the First Amino Acid of the Secreted Portion
of SEQ ID NO:Y is defined in Table 1; and an amino acid sequence of
a secreted portion of a protein encoded by a human cDNA clone
identified by a cDNA Clone Identifier in Table 1 and contained in
the deposit with the ATCC Deposit Number shown for said cDNA clone
in Table 1. The isolated polypeptide produced by this method is
also preferred.
[1099] Also preferred is a method of treatment of an individual in
need of an increased level of a secreted protein activity, which
method comprises administering to such an individual a
pharmaceutical composition comprising an amount of an isolated
polypeptide, polynucleotide, or antibody of the claimed invention
effective to increase the level of said protein activity in said
individual.
[1100] Having generally described the invention, the same will be
more readily understood by reference to the following examples,
which are provided by way of illustration and are not intended as
limiting.
EXAMPLES
Example 1
Isolation of a Selected cDNA Clone From the Deposited Sample
[1101] Each cDNA clone in a cited ATCC deposit is contained in a
plasmid vector. Table 1 identifies the vectors used to construct
the cDNA library from which each clone was isolated. In many cases,
the vector used to construct the library is a phage vector from
which a plasmid has been excised. The table immediately below
correlates the related plasmid for each phage vector used in
constructing the cDNA library. For example, where a particular
clone is identified in Table 1 as being isolated in the vector
"Lambda Zap," the corresponding deposited clone is in
"pBluescript."
68 Vector Used to Construct Library Corresponding Deposited Plasmid
Lambda Zap pBluescript (pBS) Uni-Zap XR pBluescript (pBS) Zap
Express pBK lafmid BA plafmid BA pSport1 pSport1 pCMVSport 2.0
pCMVSport 2.0 pCMVSport 3.0 pCMVSport 3.0 pCR .RTM.2.1 pCR
.RTM.2.1
[1102] Vectors Lambda Zap (U.S. Pat. Nos. 5,128,256 and 5,286,636),
Uni-Zap XR (U.S. Pat. Nos. 5,128, 256 and 5,286,636), Zap Express
(U.S. Pat. Nos. 5,128,256 and 5,286,636), pBluescript (pBS) (Short,
J. M. et al., Nucleic Acids Res. 16:7583-7600 (1988); Alting-Mees,
M. A. and Short, J. M., Nucleic Acids Res. 17:9494 (1989)) and pBK
(Alting-Mees, M. A. et al., Strategies 5:58-61 (1992)) are
commercially available from Stratagene Cloning Systems, Inc., 11011
N. Torrey Pines Road, La Jolla, Calif., 92037. pBS contains an
ampicillin resistance gene and pBK contains a neomycin resistance
gene. Both can be transformed into E. coli strain XL-1 Blue, also
available from Stratagene. pBS comes in 4 forms SK+, SK-, KS+ and
KS. The S and K refers to the orientation of the polylinker to the
T7 and T3 primer sequences which flank the polylinker region ("S"
is for SacI and "K" is for KpnI which are the first sites on each
respective end of the linker). "+" or "-" refer to the orientation
of the f1 origin of replication ("ori"), such that in one
orientation, single stranded rescue initiated from the f1 ori
generates sense strand DNA and in the other, antisense.
[1103] Vectors pSport1, pCMVSport 2.0 and pCMVSport 3.0, were
obtained from Life Technologies, Inc., P. O. Box 6009,
Gaithersburg, Md. 20897. All Sport vectors contain an ampicillin
resistance gene and may be transformed into E. coli strain DH10B,
also available from Life Technologies. (See, for instance, Gruber,
C. E., et al., Focus 15:59 (1993).) Vector lafmid BA (Bento Soares,
Columbia University, NY) contains an ampicillin resistance gene and
can be transformed into E. coli strain XL-1 Blue. Vector pCR.RTM.2.
1, which is available from Invitrogen, 1600 Faraday Avenue,
Carlsbad, Calif. 92008, contains an ampicillin resistance gene and
may be transformed into E. coli strain DH10B, available from Life
Technologies. (See, for instance, Clark, J. M., Nuc. Acids Res.
16:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991).)
Preferably, a polynucleotide of the present invention does not
comprise the phage vector sequences identified for the particular
clone in Table 1, as well as the corresponding plasmid vector
sequences designated above.
[1104] The deposited material in the sample assigned the ATCC
Deposit Number cited in Table 1 for any given cDNA clone also may
contain one or more additional plasmids, each comprising a cDNA
clone different from that given clone. Thus, deposits sharing the
same ATCC Deposit Number contain at least a plasmid for each cDNA
clone identified in Table 1. Typically, each ATCC deposit sample
cited in Table 1- comprises a mixture of approximately equal
amounts (by weight) of about 50 plasmid DNAs, each containing a
different cDNA clone; but such a deposit sample may include
plasmids for more or less than 50 cDNA clones, up to about 500 cDNA
clones.
[1105] Two approaches can be used to isolate a particular clone
from the deposited sample of plasmid DNAs cited for that clone in
Table 1. First, a plasmid is directly isolated by screening the
clones using a polynucleotide probe corresponding to SEQ ID
NO:X.
[1106] Particularly, a specific polynucleotide with 30-40
nucleotides is synthesized using an Applied Biosystems DNA
synthesizer according to the sequence reported. The oligonucleotide
is labeled, for instance, with .sup.32P-.gamma.-ATP using T4
polynucleotide kinase and purified according to routine methods.
(E.g., Maniatis et al., Molecular Cloning: A Laboratory Manual,
Cold Spring Harbor Press, Cold Spring, N.Y. (1982).) The plasmid
mixture is transformed into a suitable host, as indicated above
(such as XL-1 Blue (Stratagene)) using techniques known to those of
skill in the art, such as those provided by the vector supplier or
in related publications or patents cited above. The transformants
are plated on 1.5% agar plates (containing the appropriate
selection agent, e.g., ampicillin) to a density of about 150
transformants (colonies) per plate. These plates are screened using
Nylon membranes according to routine methods for bacterial colony
screening (e.g., Sambrook et al., Molecular Cloning: A Laboratory
Manual, 2nd Edit., (1989), Cold Spring Harbor Laboratory Press,
pages 1.93 to 1.104), or other techniques known to those of skill
in the art.
[1107] Alternatively, two primers of 17-20 nucleotides derived from
both ends of the SEQ ID NO:X (i.e., within the region of SEQ ID
NO:X bounded by the 5' NT and the 3' NT of the clone defined in
Table 1) are synthesized and used to amplify the desired cDNA using
the deposited cDNA plasmid as a template. The polymerase chain
reaction is carried out under routine conditions, for instance, in
25 .mu.l of reaction mixture with 0.5 ug of the above cDNA
template. A convenient reaction mixture is 1.5-5 mM MgCl.sub.2,
0.01% (w/v) gelatin, 20 .mu.M each of dATP, dCTP, dGTP, dTTP, 25
pmol of each primer and 0.25 Unit of Taq polymerase. Thirty five
cycles of PCR (denaturation at 94.degree. C. for 1 min; annealing
at 55.degree. C. for 1 min; elongation at 72.degree. C. for 1 min)
are performed with a Perkin-Elmer Cetus automated thermal cycler.
The amplified product is analyzed by agarose gel electrophoresis
and the DNA band with expected molecular weight is excised and
purified. The PCR product is verified to be the selected sequence
by subcloning and sequencing the DNA product.
[1108] Several methods are available for the identification of the
5' or 3' non-coding portions of a gene which may not be present in
the deposited clone. These methods include but are not limited to,
filter probing, clone enrichment using specific probes, and
protocols similar or identical to 5' and 3' "RACE" protocols which
are well known in the art. For instance, a method similar to 5'
RACE is available for generating the missing 5' end of a desired
full-length transcript. (Fromont-Racine et al., Nucleic Acids Res.
21(7):1683-1684 (1993).)
[1109] Briefly, a specific RNA oligonucleotide is ligated to the 5'
ends of a population of RNA presumably containing full-length gene
RNA transcripts. A primer set containing a primer specific to the
ligated RNA oligonucleotide and a primer specific to a known
sequence of the gene of interest is used to PCR amplify the 5'
portion of the desired full-length gene. This amplified product may
then be sequenced and used to generate the full length gene.
[1110] This above method starts with total RNA isolated from the
desired source, although poly-A+ RNA can be used. The RNA
preparation can then be treated with phosphatase if necessary to
eliminate 5' phosphate groups on degraded or damaged RNA which may
interfere with the later RNA ligase step. The phosphatase should
then be inactivated and the RNA treated with tobacco acid
pyrophosphatase in order to remove the cap structure present at the
5' ends of messenger RNAs. This reaction leaves a 5' phosphate
group at the 5' end of the cap cleaved RNA which can then be
ligated to an RNA oligonucleotide using T4 RNA ligase.
[1111] This modified RNA preparation is used as a template for
first strand cDNA synthesis using a gene specific oligonucleotide.
The first strand synthesis reaction is used as a template for PCR
amplification of the desired 5' end using a primer specific to the
ligated RNA oligonucleotide and a primer specific to the known
sequence of the gene of interest. The resultant product is then
sequenced and analyzed to confirm that the 5' end sequence belongs
to the desired gene.
Example 2
Isolation of Genomic Clones Corresponding to a Polynucleotide
[1112] A human genomic P1 library (Genomic Systems, Inc.) is
screened by PCR using primers selected for the cDNA sequence
corresponding to SEQ ID NO:X., according to the method described in
Example 1. (See also, Sambrook.)
Example 3
Tissue Distribution of Polypeptide
[1113] Tissue distribution of mRNA expression of polynucleotides of
the present invention is determined using protocols for Northern
blot analysis, described by, among others, Sambrook et al. For
example, a cDNA probe produced by the method described in Example 1
is labeled with P.sup.32 using the rediprime.TM. DNA labeling
system (Amersham Life Science), according to manufacturer's
instructions. After labeling, the probe is purified using CHROMA
SPIN-100.TM. column (Clontech Laboratories, Inc.), according to
manufacturer's protocol number PT1200-1. The purified labeled probe
is then used to examine various human tissues for mRNA
expression.
[1114] Multiple Tissue Northern (MTN) blots containing various
human tissues (H) or human immune system tissues (IM) (Clontech)
are examined with the labeled probe using ExpressHyb.TM.
hybridization solution (Clontech) according to manufacturer's
protocol number PT1190-1. Following hybridization and washing, the
blots are mounted and exposed to film at -70.degree. C. overnight,
and the films developed according to standard procedures.
Example 4
Chromosomal Mapping of the Polynucleotides
[1115] An oligonucleotide primer set is designed according to the
sequence at the 5' end of SEQ ID NO:X. This primer preferably spans
about 100 nucleotides. This primer set is then used in a polymerase
chain reaction under the following set of conditions: 30 seconds,
95.degree. C.; 1 minute, 56.degree. C.; 1 minute, 70.degree. C.
This cycle is repeated 32 times followed by one 5 minute cycle at
70.degree. C. Human, mouse, and hamster DNA is used as template in
addition to a somatic cell hybrid panel containing individual
chromosomes or chromosome fragments (Bios, Inc). The reactions is
analyzed on either 8% polyacrylamide gels or 3.5% agarose gels.
Chromosome mapping is determined by the presence of an
approximately 100 bp PCR fragment in the particular somatic cell
hybrid.
Example 5
Bacterial Expression of a Polypeptide
[1116] A polynucleotide encoding a polypeptide of the present
invention is amplified using PCR oligonucleotide primers
corresponding to the 5' and 3' ends of the DNA sequence, as
outlined in Example 1, to synthesize insertion fragments. The
primers used to amplify the cDNA insert should preferably contain
restriction sites, such as BamHI and XbaI, at the 5' end of the
primers in order to clone the amplified product into the expression
vector. For example, BamHI and XbaI correspond to the restriction
enzyme sites on the bacterial expression vector pQE-9. (Qiagen,
Inc., Chatsworth, Calif.). This plasmid vector encodes antibiotic
resistance (Amp.sup.r), a bacterial origin of replication (ori), an
IPTG-regulatable promoter/operator (P/O), a ribosome binding site
(RBS), a 6-histidine tag (6-His), and restriction enzyme cloning
sites.
[1117] The pQE-9 vector is digested with BamHI and XbaI and the
amplified fragment is ligated into the pQE-9 vector maintaining the
reading frame initiated at the bacterial RBS. The ligation mixture
is then used to transform the E. coli strain M15/rep4 (Qiagen,
Inc.) which contains multiple copies of the plasmid pREP4, which
expresses the lacI repressor and also confers kanamycin resistance
(Kan.sup.r). Transformants are identified by their ability to grow
on LB plates and ampicillin/kanamycin resistant colonies are
selected. Plasmid DNA is isolated and confirmed by restriction
analysis.
[1118] Clones containing the desired constructs are grown overnight
(O/N) in liquid culture in LB media supplemented with both Amp (100
ug/ml) and Kan (25 ug/ml). The O/N culture is used to inoculate a
large culture at a ratio of 1:100 to 1:250. The cells are grown to
an optical density 600 (O.D..sup.600) of between 0.4 and 0.6. IPTG
(Isopropyl-B-D-thiogalacto pyranoside) is then added to a final
concentration of 1 mM. IPTG induces by inactivating the lacI
repressor, clearing the P/O leading to increased gene
expression.
[1119] Cells are grown for an extra 3 to 4 hours. Cells are then
harvested by centrifugation (20 mins at 6000.times. g). The cell
pellet is solubilized in the chaotropic agent 6 Molar Guanidine HCl
by stirring for 3-4 hours at 4.degree. C. The cell debris is
removed by centrifugation, and the supernatant containing the
polypeptide is loaded onto a nickel-nitrilo-tri-acetic acid
("Ni-NTA") affinity resin column (available from QIAGEN, Inc.,
supra). Proteins with a 6.times. His tag bind to the Ni-NTA resin
with high affinity and can be purified in a simple one-step
procedure (for details see: The QIAexpressionist (1995) QIAGEN,
Inc., supra).
[1120] Briefly, the supernatant is loaded onto the column in 6 M
guanidine-HCl, pH 8, the column is first washed with 10 volumes of
6 M guanidine-HCl, pH 8, then washed with 10 volumes of 6 M
guanidine-HCl pH 6, and finally the polypeptide is eluted with 6 M
guanidine-HCl, pH 5.
[1121] The purified protein is then renatured by dialyzing it
against phosphate-buffered saline (PBS) or 50 mM Na-acetate, pH 6
buffer plus 200 mM NaCl. Alternatively, the protein can be
successfully refolded while immobilized on the Ni-NTA column. The
recommended conditions are as follows: renature using a linear
6M-1M urea gradient in 500 mM NaCl, 20% glycerol, 20 mM Tris/HCl pH
7.4, containing protease inhibitors. The renaturation should be
performed over a period of 1.5 hours or more. After renaturation
the proteins are eluted by the addition of 250 mM immidazole.
Immidazole is removed by a final dialyzing step against PBS or 50
mM sodium acetate pH 6 buffer plus 200 mM NaCl. The purified
protein is stored at 4.degree. C. or frozen at -80.degree. C.
[1122] In addition to the above expression vector, the present
invention further includes an expression vector comprising phage
operator and promoter elements operatively linked to a
polynucleotide of the present invention, called pHE4a. (ATCC
Accession Number 209645, deposited on February 25, 1998.) This
vector contains: 1) a neomycinphosphotransferase gene as a
selection marker, 2) an E. coli origin of replication, 3) a T5
phage promoter sequence, 4) two lac operator sequences, 5) a
Shine-Delgarno sequence, and 6) the lactose operon repressor gene
(lacIq). The origin of replication (oriC) is derived from pUC19
(LTI, Gaithersburg, Md.). The promoter sequence and operator
sequences are made synthetically.
[1123] DNA can be inserted into the pHEa by restricting the vector
with NdeI and XbaI, BamHI, XhoI, or Asp718, running the restricted
product on a gel, and isolating the larger fragment (the stuffer
fragment should be about 310 base pairs). The DNA insert is
generated according to the PCR protocol described in Example 1,
using PCR primers having restriction sites for NdeI (5' primer) and
XbaI, BamHI, XhoI, or Asp718 (3' primer). The PCR insert is gel
purified and restricted with compatible enzymes. The insert and
vector are ligated according to standard protocols.
[1124] The engineered vector could easily be substituted in the
above protocol to express protein in a bacterial system.
Example 6
Purification of a Polypeptide from an Inclusion Body
[1125] The following alternative method can be used to purify a
polypeptide expressed in E coli when it is present in the form of
inclusion bodies. Unless otherwise specified, all of the following
steps are conducted at 4-10.degree. C.
[1126] Upon completion of the production phase of the E. coli
fermentation, the cell culture is cooled to 4-10.degree. C. and the
cells harvested by continuous centrifugation at 15,000 rpm (Heraeus
Sepatech). On the basis of the expected yield of protein per unit
weight of cell paste and the amount of purified protein required,
an appropriate amount of cell paste, by weight, is suspended in a
buffer solution containing 100 mM Tris, 50 mM EDTA, pH 7.4. The
cells are dispersed to a homogeneous suspension using a high shear
mixer.
[1127] The cells are then lysed by passing the solution through a
microfluidizer (Microfuidics, Corp. or APV Gaulin, Inc.) twice at
4000-6000 psi. The homogenate is then mixed with NaCl'solution to a
final concentration of 0.5 M NaCl, followed by centrifugation at
7000.times. g for 15 min. The resultant pellet is washed again
using 0.5M NaCl, 100 mM Tris, 50 mM EDTA, pH 7.4.
[1128] The resulting washed inclusion bodies are solubilized with
1.5 M guanidine hydrochloride (GuHCl) for 2-4 hours. After
7000.times. g centrifugation for 15 min., the pellet is discarded
and the polypeptide containing supernatant is incubated at
4.degree. C. overnight to allow further GuHCl extraction.
[1129] Following high speed centrifugation (30,000.times. g) to
remove insoluble particles, the GuHCl solubilized protein is
refolded by quickly mixing the GuHCl extract with 20 volumes of
buffer containing 50 mM sodium, pH 4.5, 150 mM NaCl, 2 mM EDTA by
vigorous stirring. The refolded diluted protein solution is kept at
4.degree. C. without mixing for 12 hours prior to further
purification steps.
[1130] To clarify the refolded polypeptide solution, a previously
prepared tangential filtration unit equipped with 0.16 .mu.m
membrane filter with appropriate surface area (e.g., Filtron),
equilibrated with 40 mM sodium acetate, pH 6.0 is employed. The
filtered sample is loaded onto a cation exchange resin (e.g., Poros
HS-50, Perseptive Biosystems). The column is washed with 40 mM
sodium acetate, pH 6.0 and eluted with 250 mM, 500 mM, 1000 mM, and
1500 mM NaCl in the same buffer, in a stepwise manner. The
absorbance at 280 nm of the effluent is continuously monitored.
Fractions are collected and further analyzed by SDS-PAGE.
[1131] Fractions containing the polypeptide are then pooled and
mixed with 4 volumes of water. The diluted sample is then loaded
onto a previously prepared set of tandem columns of strong anion
(Poros HQ-50, Perseptive Biosystems) and weak anion (Poros CM-20,
Perseptive Biosystems) exchange resins. The columns are
equilibrated with 40 mM sodium acetate, pH 6.0. Both columns are
washed with 40 MM sodium acetate, pH 6.0, 20.0 mM NaCl. The CM-20
column is then eluted using a 10 column volume linear gradient
ranging from 0.2 M NaCl, 50 mM sodium acetate, pH 6.0 to 1.0 M
NaCl, 50 mM sodium acetate, pH 6.5. Fractions are collected under
constant A.sub.280 monitoring of the effluent. Fractions containing
the polypeptide (determined, for instance, by 16% SDS-PAGE) are
then pooled.
[1132] The resultant polypeptide should exhibit greater than 95%
purity after the above refolding and purification steps. No major
contaminant bands should be observed from Commassie blue stained
16% SDS-PAGE gel when 5 .mu.g of purified protein is loaded. The
purified protein can also be tested for endotoxinlLPS
contamination, and typically the LPS content is less than 0.1 ng/ml
according to LAL assays.
Example 7
Cloning and Expression of a Polypeptide in a Baculovirus Expression
System
[1133] In this example, the plasmid shuttle vector pA2 is used to
insert a polynucleotide into a baculovirus to express a
polypeptide. This expression vector contains the strong polyhedrin
promoter of the Autographa californica nuclear polyhedrosis virus
(AcMNPV) followed by convenient restriction sites such as BamHI,
XbaI and Asp718. The polyadenylation site of the simian virus 40
("SV40") is used for efficient polyadenylation. For easy selection
of recombinant virus, the plasmid contains the beta-galactosidase
gene from E. coli under control of a weak Drosophila promoter in
the same orientation, followed by the polyadenylation signal of the
polyhedrin gene. The inserted genes are flanked on both sides by
viral sequences for cell-mediated homologous recombination with
wild-type viral DNA to generate a viable virus that express the
cloned polynucleotide.
[1134] Many other baculovirus vectors can be used in place of the
vector above, such as pAc373, pVL941, and pAcIM1, as one skilled in
the art would readily appreciate, as long as the construct provides
appropriately located signals for transcription, translation,
secretion and the like, including a signal peptide and an in-frame
AUG as required. Such vectors are described, for instance, in
Luckow et al., Virology 170:31-39 (1989).
[1135] Specifically, the cDNA sequence contained in the deposited
clone, including the AUG initiation codon and the naturally
associated leader sequence identified in Table 1, is amplified
using the PCR protocol described in Example 1. If the naturally
occurring signal sequence is used to produce the secreted protein,
the pA2 vector does not need a second signal peptide.
Alternatively, the vector can be modified (pA2 GP) to include a
baculovirus leader sequence, using the standard methods described
in Summers et al., "A Manual of Methods for Baculovirus Vectors and
Insect Cell Culture Procedures," Texas Agricultural Experimental
Station Bulletin No. 1555 (1987).
[1136] The amplified fragment is isolated from a 1% agarose gel
using a commercially available kit ("Geneclean," BIO 101 Inc., La
Jolla, Calif.). The fragment then is digested with appropriate
restriction enzymes and again purified on a 1% agarose gel.
[1137] The plasmid is digested with the corresponding restriction
enzymes and optionally, can be dephosphorylated using calf
intestinal phosphatase, using routine procedures known in the art.
The DNA is then isolated from a 1% agarose gel using a commercially
available kit ("Geneclean" BIO 101 Inc., La Jolla, Calif.).
[1138] The fragment and the dephosphorylated plasmid are ligated
together with T4 DNA ligase. E. coli HB 101 or other suitable E.
coli hosts such as XL-1 Blue (Stratagene Cloning Systems, La Jolla,
Calif.) cells are transformed with the ligation mixture and spread
on culture plates. Bacteria containing the plasmid are identified
by digesting DNA from individual colonies and analyzing the
digestion product by gel electrophoresis. The sequence of the
cloned fragment is confirmed by DNA sequencing.
[1139] Five .mu.g of a plasmid containing the polynucleotide is
co-transfected with 1.0 .mu.g of a commercially available
linearized baculovirus DNA ("BaculoGold.TM. baculovirus DNA",
Pharmingen, San Diego, Calif.), using the lipofection method
described by Felgner et al., Proc. Natl. Acad. Sci. USA
84:7413-7417 (1987). One .mu.g of BaculoGold.TM. virus DNA and 5
.mu.g of the plasmid are mixed in a sterile well of a microtiter
plate containing 50 .mu.l of serum-free Grace's medium (Life
Technologies Inc., Gaithersburg, Md.). Afterwards, 10 .mu.l
Lipofectin plus 90 .mu.l Grace's medium are added, mixed and
incubated for 15 minutes at room temperature. Then the transfection
mixture is added drop-wise to Sf9 insect cells (ATCC CRL 1711)
seeded in a 35 mm tissue culture plate with 1 ml Grace's medium
without serum. The plate is then incubated for 5 hours at
27.degree. C. The transfection solution is then removed from the
plate and 1 ml of Grace's insect medium supplemented with 10% fetal
calf serum is added. Cultivation is then continued at 27.degree. C.
for four days.
[1140] After four days the supernatant is collected and a plaque
assay is performed, as described by Summers and Smith, supra. An
agarose gel with "Blue Gal" (Life Technologies Inc., Gaithersburg)
is used to allow easy identification and isolation of
gal-expressing clones, which produce blue-stained plaques. (A
detailed description of a "plaque assay" of this type can also be
found in the user's guide for insect cell culture and
baculovirology distributed by Life Technologies Inc., Gaithersburg,
page 9-10.) After appropriate incubation, blue stained plaques are
picked with the tip of a micropipettor (e.g., Eppendorf). The agar
containing the recombinant viruses is then resuspended in a
microcentrifuge tube containing 200 .mu.l of Grace's medium and the
suspension containing the recombinant baculovirus is used to infect
Sf9 cells seeded in 35 mm dishes. Four days later the supernatants
of these culture dishes are harvested and then they are stored at
4.degree. C.
[1141] To verify the expression of the polypeptide, Sf9 cells are
grown in Grace's medium supplemented with 10% heat-inactivated FBS.
The cells are infected with the recombinant baculovirus containing
the polynucleotide at a multiplicity of infection ("MOI") of about
2. If radiolabeled proteins are desired, 6 hours later the medium
is removed and is replaced with SF900 II medium minus methionine
and cysteine (available from Life Technologies Inc., Rockville,
Md.). After 42 hours, 5 .mu.Ci of .sup.35S-methionine and 5 .mu.Ci
.sup.35S-cysteine (available from Amersham) are added. The cells
are further incubated for 16 hours and then are harvested by
centrifugation. The proteins in the supernatant as well as the
intracellular proteins are analyzed by SDS-PAGE followed by
autoradiography (if radiolabeled).
[1142] Microsequencing of the amino acid sequence of the amino
terminus of purified protein may be used to determine the amino
terminal sequence of the produced protein.
Example 8
Expression of a Polypeptide in Mammalian Cells
[1143] The polypeptide of the present invention can be expressed in
a mammalian cell. A typical mammalian expression vector contains a
promoter element, which mediates the initiation of transcription of
mRNA, a protein coding sequence, and signals required for the
termination of transcription and polyadenylation of the transcript.
Additional elements include enhancers, Kozak sequences and
intervening sequences flanked by donor and acceptor sites for RNA
splicing. Highly efficient transcription is achieved with the early
and late promoters from SV40, the long terminal repeats (LTRs) from
Retroviruses, e.g., RSV, HTLVI, HIVI and the early promoter of the
cytomegalovirus (CMV). However, cellular elements can also be used
(e.g., the human actin promoter).
[1144] Suitable expression vectors for use in practicing the
present invention include, for example, vectors such as pSVL and
pMSG (Pharmacia, Uppsala, Sweden), pRSVcat (ATCC 37152), pSV2dhfr
(ATCC 37146), pBC12MI (ATCC 67109), pCMVSport 2.0, and pCMVSport
3.0. Mammalian host cells that could be used include, human Hela,
293, H9 and Jurkat cells, mouse NIH3T3 and C127 cells, Cos 1, Cos 7
and CV1, quail QC1-3 cells, mouse L cells and Chinese hamster ovary
(CHO) cells.
[1145] Alternatively, the polypeptide can be expressed in stable
cell lines containing the polynucleotide integrated into a
chromosome. The co-transfection with a selectable marker such as
dhfr, gpt, neomycin, hygromycin allows the identification and
isolation of the transfected cells.
[1146] The transfected gene can also be amplified to express large
amounts of the encoded protein. The DHFR (dihydrofolate reductase)
marker is useful in developing cell lines that carry several
hundred or even several thousand copies of the gene of interest.
(See, e.g., Alt, F. W., et al., J. Biol. Chem. 253:1357-1370
(1978); Hamlin, J. L. and Ma, C., Biochem. et Biophys. Acta,
1097:107-143 (1990); Page, M. J. and Sydenham, M. A., Biotechnology
9:64-68 (1991).) Another useful selection marker is the enzyme
glutamine synthase (GS) (Murphy et al., Biochem J. 227:277-279
(1991); Bebbington et al., Bio/Technology 10:169-175 (1992). Using
these markers, the mammalian cells are grown in selective medium
and the cells with the highest resistance are selected. These cell
lines contain the amplified gene(s) integrated into a chromosome.
Chinese hamster ovary (CHO) and NSO cells are often used for the
production of proteins.
[1147] Derivatives of the plasmid pSV2-dhfr (ATCC Accession No.
37146), the expression vectors pC4 (ATCC Accession No. 209646) and
pC6 (ATCC Accession No.209647) contain the strong promoter (LTR) of
the Rous Sarcoma Virus (Cullen et al., Molecular and Cellular
Biology, 438-447 (March, 1985)) plus a fragment of the CMV-enhancer
(Boshart et al., Cell 41:521-530 (1985).) Multiple cloning sites,
e.g., with the restriction enzyme cleavage sites BamHI, XbaI and
Asp718, facilitate the cloning of the gene of interest. The vectors
also contain the 3' intron, the polyadenylation and termination
signal of the rat preproinsulin gene, and the mouse DHFR gene under
control of the SV40 early promoter.
[1148] Specifically, the plasmid pC6, for example, is digested with
appropriate restriction enzymes and then dephosphorylated using
calf intestinal phosphates by procedures known in the art. The
vector is then isolated from a 1% agarose gel.
[1149] A polynucleotide of the present invention is amplified
according to the protocol outlined in Example 1. If the naturally
occurring signal sequence is used to produce the secreted protein,
the vector does not need a second signal peptide. Alternatively, if
the naturally occurring signal sequence is not used, the vector can
be modified to include a heterologous signal sequence. (See, e.g.,
WO 96/34891.)
[1150] The amplified fragment is isolated from a 1% agarose gel
using a commercially available kit ("Geneclean," BIO 101 Inc., La
Jolla, Calif.). The fragment then is digested with appropriate
restriction enzymes and again purified on a 1% agarose gel.
[1151] The amplified fragment is then digested with the same
restriction enzyme and purified on a 1% agarose gel. The isolated
fragment and the dephosphorylated vector are then ligated with T4
DNA ligase. E. coli HB 101 or XL-1 Blue cells are then transformed
and bacteria are identified that contain the fragment inserted into
plasmid pC6 using, for instance, restriction enzyme analysis.
[1152] Chinese hamster ovary cells lacking an active DHFR gene is
used for transfection. Five .mu.g of the expression plasmid pC6 is
cotransfected with 0.5 .mu.g of the plasmid pSVneo using lipofectin
(Felgner et al., supra). The plasmid pSV2-neo contains a dominant
selectable marker, the neo gene from Tn5 encoding an enzyme that
confers resistance to a group of antibiotics including G418. The
cells are seeded in alpha minus MEM supplemented with 1 mg/ml G418.
After 2 days, the cells are trypsinized and seeded in hybridoma
cloning plates (Greiner, Germany) in alpha minus MEM supplemented
with 10, 25, or 50 ng/ml of metothrexate plus 1 mg/ml G418. After
about 10-14 days single clones are trypsinized and then seeded in
6-well petri dishes or 10 ml flasks using different concentrations
of methotrexate (50 nM, 100 nM, 200 nM, 400 nM, 800 nM). Clones
growing at the highest concentrations of methotrexate are then
transferred to new 6-well plates containing even higher
concentrations of methotrexate (1 .mu.M, 2 .mu.M, 5 .mu.M, 10 mM,
20 mM). The same procedure is repeated until clones are obtained
which grow at a concentration of 100-200 .mu.M. Expression of the
desired gene product is analyzed, for instance, by SDS-PAGE and
Western blot or by reversed phase HPLC analysis.
Example 9
Protein Fusions
[1153] The polypeptides of the present invention are preferably
fused to other proteins. These fusion proteins can be used for a
variety of applications. For example, fusion of the present
polypeptides to His-tag, HA-tag, protein A, IgG domains, and
maltose binding protein facilitates purification. (See Example 5;
see also EP A 394,827; Traunecker, et al., Nature 331:84-86
(1988).) Similarly, fusion to IgG-1, IgG-3, and albumin increases
the halflife time in vivo. Nuclear localization signals fused to
the polypeptides of the present invention can target the protein to
a specific subcellular localization, while covalent heterodimer or
homodimers can increase or decrease the activity of a fusion
protein. Fusion proteins can also create chimeric molecules having
more than one function. Finally, fusion proteins can increase
solubility and/or stability of the fused protein compared to the
non-fused protein. All of the types of fusion proteins described
above can be made by modifying the following protocol, which
outlines the fusion of a polypeptide to an IgG molecule, or the
protocol described in Example 5.
[1154] Briefly, the human Fc portion of the IgG molecule can be PCR
amplified, using primers that span the 5' and 3' ends of the
sequence described below. These primers also should have convenient
restriction enzyme sites that will facilitate cloning into an
expression vector, preferably a mammalian expression vector.
[1155] For example, if pC4 (Accession No. 209646) is used, the
human Fc portion can be ligated into the BamHI cloning site. Note
that the 3' BamHI site should be destroyed. Next, the vector
containing the human Fc portion is re-restricted with BamHI,
linearizing the vector, and a polynucleotide of the present
invention, isolated by the PCR protocol described in Example 1, is
ligated into this BamHI site. Note that the polynucleotide is
cloned without a stop codon, otherwise a fusion protein will not be
produced.
[1156] If the naturally occurring signal sequence is used to
produce the secreted protein, pC4 does not need a second signal
peptide. Alternatively, if the naturally occurring signal sequence
is not used, the vector can be modified to include a heterologous
signal sequence. (See, e.g., WO 96/34891.)
[1157] Human IgG Fc Region:
69 GGGATCCGGAGCCCAAATCTTCTGACAAAACTCACA (SEQ ID NO: 1)
CATGCCCACCGTGCCCAGCACCTGAATTCGAGGGTG
CACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGG
ACACCCTCATGATCTCCCGGACTCCTGAGGTCACAT
GCGTGGTGGTGGACGTAAGCCACGAAGACCCTGAGG
TCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGC
ATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACA
ACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCC
TGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGT
GCAAGGTCTCCAACAAAGCCCTCCCAACCCCCATCG
AGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAG
AACCACAGGTGTACACCCTGCCCCCATCCCGGGATG
AGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGG
TCAAAGGCTTCTATCCAAGCGACATCGCCGTGGAGT
GGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGA
CCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCT
TCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGT
GGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGC
ATGAGGCTCTGCACAACCACTACACGCAGAAGAGCC
TCTCCCTGTCTCCGGGTAAATGAGTGCGACGGCCGC GACTCTAGAGGAT
Example 10
Production of an Antibody from a Polypeptide
[1158] The antibodies of the present invention can be prepared by a
variety of methods. (See, Current Protocols, Chapter 2.) For
example, cells expressing a polypeptide of the present invention is
administered to an animal to induce the production of sera
containing polyclonal antibodies. In a preferred method, a
preparation of the secreted protein is prepared and purified to
render it substantially free of natural contaminants. Such a
preparation is then introduced into an animal in order to produce
polyclonal antisera of greater specific activity.
[1159] In the most preferred method, the antibodies of the present
invention are monoclonal antibodies (or protein binding fragments
thereof). Such monoclonal antibodies can be prepared using
hybridoma technology. (Kohler et al., Nature 256:495 (1975); Kohler
et al., Eur. J. Immunol. 6:511 (1976); Kohler et al., Eur. J.
Immunol. 6:292 (1976); Hammerling et al., in: Monoclonal Antibodies
and T-Cell Hybridomas, Elsevier, N.Y., pp. 563-681 (1981).) In
general, such procedures involve immunizing an animal (preferably a
mouse) with polypeptide or, more preferably, with a secreted
polypeptide-expressing cell. Such cells may be cultured in any
suitable tissue culture medium; however, it is preferable to
culture cells in Earle's modified Eagle's medium supplemented with
10% fetal bovine serum (inactivated at about 56.degree. C.), and
supplemented with about 10 g/l of nonessential amino acids, about
1,000 U/ml of penicillin, and about 100 .mu.g/ml of
streptomycin.
[1160] The splenocytes of such mice are extracted and fused with a
suitable myeloma cell line. Any suitable myeloma cell line may be
employed in accordance with the present invention; however, it is
preferable to employ the parent myeloma cell line (SP20), available
from the ATCC. After fusion, the resulting hybridoma cells are
selectively maintained in HAT medium, and then cloned by limiting
dilution as described by Wands et al. (Gastroenterology 80:225-232
(1981).) The hybridoma cells obtained through such a selection are
then assayed to identify clones which secrete antibodies capable of
binding the polypeptide.
[1161] Alternatively, additional antibodies capable of binding to
the polypeptide can be produced in a two-step procedure using
anti-idiotypic antibodies. Such a method makes use of the fact that
antibodies are themselves antigens, and therefore, it is possible
to obtain an antibody which binds to a second antibody. In
accordance with this method, protein specific antibodies are used
to immunize an animal, preferably a mouse. The splenocytes of such
an animal are then used to produce hybridoma cells, and the
hybridoma cells are screened to identify clones which produce an
antibody whose ability to bind to the protein-specific antibody can
be blocked by the polypeptide. Such antibodies comprise
anti-idiotypic antibodies to the protein-specific antibody and can
be used to immunize an animal to induce formation of further
protein-specific antibodies.
[1162] It will be appreciated that Fab and F(ab')2 and other
fragments of the antibodies of the present invention may be used
according to the methods disclosed herein. Such fragments are
typically produced by proteolytic cleavage, using enzymes such as
papain (to produce Fab fragments) or pepsin (to produce F(ab')2
fragments). Alternatively, secreted protein-binding fragments can
be produced through the application of recombinant DNA technology
or through synthetic chemistry.
[1163] For in vivo use of antibodies in humans, it may be
preferable to use "humanized" chimeric monoclonal antibodies. Such
antibodies can be produced using genetic constructs derived from
hybridoma cells producing the monoclonal antibodies described
above. Methods for producing chimeric antibodies are known in the
art. (See, for review, Morrison, Science 229:1202 (1985); Oi et
al., BioTechniques 4:214 (1986); Cabilly et al., U.S. Pat. No.
4,816,567; Taniguchi et al., EP 171496; Morrison et al., EP 173494;
Neuberger et al., WO 8601533; Robinson et al., WO 8702671;
Boulianne et al., Nature 312:643 (1984); Neubergeret al., Nature
314:268 (1985).)
Example 11
Production Of Secreted Protein For High-throughput Screening
Assays
[1164] The following protocol produces a supernatant containing a
polypeptide to be tested. This supernatant can then be used in the
Screening Assays described in Examples 13-20.
[1165] First, dilute Poly-D-Lysine (644 587 Boehringer-Mannheim)
stock solution (1 mg/ml in PBS) 1:20 in PBS (w/o calcium or
magnesium 17-516F Biowhittaker) for a working solution of 50 ug/ml.
Add 200 ul of this solution to each well (24 well plates) and
incubate at RT for 20 minutes. Be sure to distribute the solution
over each well (note: a 12-channel pipetter may be used with tips
on every other channel). Aspirate off the Poly-D-Lysine solution
and rinse with 1 ml PBS (Phosphate Buffered Saline). The PBS should
remain in the well until just prior to plating the cells and plates
may be poly-lysine coated in advance for up to two weeks.
[1166] Plate 293T cells (do not carry cells past P+20) at
2.times.10.sup.5 cells/well in 0.5 ml DMEM(Dulbecco's Modified
Eagle Medium)(with 4.5 G/L glucose and L-glutamine (12-604F
Biowhittaker))/10% heat inactivated FBS(14-503F
Biowhittaker)/1.times. Penstrep(17-602E Biowhittaker). Let the
cells grow overnight.
[1167] The next day, mix together in a sterile solution basin: 300
ul Lipofectamine (18324-012 Gibco/BRL) and 5 ml Optimem I (31985070
Gibco/BRL)/96-well plate. With a small volume multi-channel
pipetter, aliquot approximately 2 ug of an expression vector
containing a polynucleotide insert, produced by the methods
described in Examples 8 or 9, into an appropriately labeled 96-well
round bottom plate. With a multi-channel pipetter, add 50 ul of the
Lipofectamine/Optimem I mixture to each well. Pipette up and down
gently to mix. Incubate at RT 15-45 minutes. After about 20
minutes, use a multi-channel pipetter to add 150 ul Optimem I to
each well. As a control, one plate of vector DNA lacking an insert
should be transfected with each set of transfections.
[1168] Preferably, the transfection should be performed by
tag-teaming the following tasks. By tag-teaming, hands on time is
cut in half, and the cells do not spend too much time on PBS.
First, person A aspirates off the media from four 24-well plates of
cells, and then person B rinses each well with 0.5-1 ml PBS. Person
A then aspirates off PBS rinse, and person B, using a 12-channel
pipetter with tips on every other channel, adds the 200 ul of
DNA/Lipofectamine/Optimem I complex to the odd wells first, then to
the even wells, to each row on the 24-well plates. Incubate at
37.degree. C. for 6 hours.
[1169] While cells are incubating, prepare appropriate media,
either 1%BSA in DMEM with 1.times. penstrep, or CHO-5 media (116.6
mg/L of CaCl2 (anhyd); 0.00130 mg/L CuSO.sub.4-5H.sub.2O; 0.050
mg/L of Fe(NO.sub.3).sub.3-9H.sub.2O; 0.417 mg/L of
FeSO.sub.4-7H.sub.2O; 311.80 mg/L of Kcl; 28.64 mg/L of MgCl.sub.2;
48.84 mg/L of MgSO.sub.4; 6995.50 mg/L of NaCl; 2400.0 mg/L of
NaHCO.sub.3; 62.50 mg/L of NaH.sub.2PO.sub.4--H.sub.2O; 71.02 mg/L
of NaHPO4; 0.4320 mg/L of ZnSO.sub.4-7H.sub.2O; 0.002 mg/L of
Arachidonic Acid; 1.022 mg/L of Cholesterol; 0.070 mg/L of
DL-alpha-Tocopherol-Acetate; 0.0520 mg/L of Linoleic Acid; 0.010
mg/L of Linolenic Acid; 0.010 mg/L of Myristic Acid; 0.010 mg/L of
Oleic Acid; 0.010 mg/L of Palmitric Acid; 0.010 mg/L of Palmitic
Acid; 100 mg/L of Pluronic F-68; 0.010 mg/L of Stearic Acid; 2.20
mg/L of Tween 80; 4551 mg/L of D-Glucose; 130.85 mg/ml of
L-Alanine; 147.50 mg/ml of L-Arginine-HCL; 7.50 mg/ml of
L-Asparagine-H.sub.2O; 6.65 mg/ml of L-Aspartic Acid; 29.56 mg/ml
of L-Cystine-2HCL-H.sub.2O; 31.29 mg/ml of L-Cystine-2HCL; 7.35
mg/ml of L-Glutamic Acid; 365.0 mg/ml of L-Glutamine; 18.75 mg/ml
of Glycine; 52.48 mg/ml of L-Histidine-HCL-H.sub.2O; 106.97 mg/ml
of L-Isoleucine; 111.45 mg/ml of L-Leucine; 163.75 mg/ml of
L-Lysine HCL; 32.34 mg/ml of L-Methionine; 68.48 mg/ml of
L-Phenylalamine; 40.0 mg/ml of L-Proline; 26.25 mg/ml of L-Serine;
101.05 mg/ml of L-Threonine; 19.22 mg/ml of L-Tryptophan; 91.79
mg/ml of L-Tryrosine-2Na-2H.sub.2O; 99.65 mg/ml of L-Valine; 0.0035
mg/L of Biotin; 3.24 mg/L of D-Ca Pantothenate; 11.78 mg/L of
Choline Chloride; 4.65 mg/L of Folic Acid; 15.60 mg/L of
i-Inositol; 3.02 mg/L of Niacinamide; 3.00 mg/L of Pyridoxal HCL;
0.031 mg/L of Pyridoxine HCL; 0.319 mg/L of Riboflavin; 3.17 mg/L
of Thiamine HCL; 0.365 mg/L of Thymidine; and 0.680 mg/L of Vitamin
B.sub.12; 25 mM of HEPES Buffer; 2.39 mg/L of Na Hypoxanthine;
0.105 mg/L of Lipoic Acid; 0.081 mg/L of Sodium Putrescine-2HCL;
55.0 mg/L of Sodium Pyruvate; 0.0067 mg/L of Sodium Selenite; 20 uM
of Ethanolamine; 0.122 mg/L of Ferric Citrate; 41.70 mg,/L of
Methyl-B-Cyclodextrin complexed with Linoleic Acid; 33.33 mg/L of
Methyl-B-Cyclodextrin complexed with Oleic Acid; and 10 mg/L of
Methyl-B-Cyclodextrin complexed with Retinal) with 2 mm glutamine
and 1.times. penstrep. (BSA (81-068-3 Bayer) 100 .mu.m dissolved in
1 L DMEM for a 10% BSA stock solution). Filter the media and
collect 50 ul for endotoxin assay in 15 ml polystyrene conical.
[1170] The transfection reaction is terminated, preferably by
tag-teaming, at the end of the incubation period. Person A
aspirates off the transfection media, while person B adds 1.5 ml
appropriate media to each well. Incubate at 37.degree. C. for 45 or
72 hours depending on the media used: 1%BSA for 45 hours or CHO-5
for 72 hours.
[1171] On day four, using a 300 ul multichannel pipetter, aliquot
600 ul in one 1 ml deep well plate and the remaining supernatant
into a 2 ml deep well. The supernatants from each well can then be
used in the assays described in Examples 13-20.
[1172] It is specifically understood that when activity is obtained
in any of the assays described below using a supernatant, the
activity originates from either the polypeptide directly (e.g., as
a secreted protein) or by the polypeptide inducing expression of
other proteins, which are then secreted into the supernatant. Thus,
the invention further provides a method of identifying the protein
in the supernatant characterized by an activity in a particular
assay.
Example 12
Construction of GAS Reporter Construct
[1173] One signal transduction pathway involved in the
differentiation and proliferation of cells is called the Jaks-STATs
pathway. Activated proteins in the Jaks-STATs pathway bind to gamma
activation site "GAS" elements or interferon-sensitive responsive
element ("ISRE"), located in the promoter of many genes. The
binding of a protein to these elements alter the expression of the
associated gene.
[1174] GAS and ISRE elements are recognized by a class of
transcription factors called Signal Transducers and Activators of
Transcription, or "STATs." There are six members of the STATs
family. Stat1 and Stat3 are present in many cell types, as is Stat2
(as response to IFN-alpha is widespread). Stat4 is more restricted
and is not in many cell types though it has been found in T helper
class I, cells after treatment with IL-12. Stat5 was originally
called mammary growth factor, but has been found at higher
concentrations in other cells including myeloid cells. It can be
activated in tissue culture cells by many cytokines.
[1175] The STATs are activated to translocate from the cytoplasm to
the nucleus upon tyrosine phosphorylation by a set of kinases known
as the Janus Kinase ("Jaks") family. Jaks represent a distinct
family of soluble tyrosine kinases and include Tyk2, Jak1, Jak2,
and Jak3. These kinases display significant sequence similarity and
are generally catalytically inactive in resting cells.
[1176] The Jaks are activated by a wide range of receptors
summarized in the Table below. (Adapted from review by Schidler and
Darnell, Ann. Rev. Biochem. 64:621-51 (1995).) A cytokine receptor
family, capable of activating Jaks, is divided into two groups: (a)
Class 1 includes receptors for IL-2, IL-3, IL-4, IL-6, IL-7, IL-9,
IL-11, IL-12, IL-15, Epo, PRL, GH, G-CSF, GM-CSF, LIF, CNTF, and
thrombopoietin; and
[1177] (b) Class 2 includes IFN-a, IFN-g, and IL-10. The Class 1
receptors share a conserved cysteine motif (a set of four conserved
cysteines and one tryptophan) and a WSXWS motif (a membrane
proximal region encoding Trp-Ser-Xxx-Trp-Ser (SEQ ID NO:2)).
[1178] Thus, on binding of a ligand to a receptor, Jaks are
activated, which in turn activate STATs, which then translocate and
bind to GAS elements. This entire process is encompassed in the
Jaks-STATs signal transduction pathway.
[1179] Therefore, activation of the Jaks-STATs pathway, reflected
by the binding of the GAS or the ISRE element, can be used to
indicate proteins involved in the proliferation and differentiation
of cells. For example, growth factors and cytokines are known to
activate the Jaks-STATs pathway. (See Table below.) Thus, by using
GAS elements linked to reporter molecules, activators of the
Jaks-STATs pathway can be identified.
70 JAKs Ligand tyk2 Jak1 Jak2 Jak3 STATS GAS(elements) or ISRE IFN
family IFN-a/B + + - - 1,2,3 ISRE IFN-g + + - 1 GAS
(IRF1>Lys6>IFP) Il-10 + ? ? - 1,3 gp130 family IL-6
(Pleiotrophic) + + + ? 1,3 GAS (IRF1>Lys6>IFP)
Il-11(Pleiotrophic) ? + ? ? 1,3 OnM(Pleiotrophic) ? + + ? 1,3
LIF(Pleiotrophic) ? + + ? 1,3 CNTF(Pleiotrophic) -/+ + + ? 1,3
G-CSF(Pleiotrophic) ? + ? ? 1,3 IL-12(Pleiotrophic) + - + + 1,3 g-C
family IL-2 (lymphocytes) - + - + 1,3,5 GAS IL-4 (lymph/myeloid) -
+ - + 6 GAS (IRF1 = IFP >>Ly6)(IgH) IL-7 (lymphocytes) - + -
+ 5 GAS IL-9 (lymphocytes) - + - + 5 GAS IL-13 (lymphocyte) - + ? ?
6 GAS IL-15 ? + ? + 5 GAS gp140 family IL-3 (myeloid) - - + - 5 GAS
(IRF1>IFP>>Ly6) IL-5 (myeloid) - - + - 5 GAS GM-CSF
(myeloid) - - + - 5 GAS Growth hormone family GH ? - + - 5 PRL ?
+/- + - 1,3,5 EPO ? - + - 5 GAS(B-CAS>IRF1=IFP>>Ly6)
Receptor Tyrosine Kinases EGF ? + + - 1,3 GAS(IRF1) PDGF ? + + -
1,3 CSF-1 ? + + - 1,3 GAS (not IRF1)
[1180] To construct a synthetic GAS containing promoter element,
which is used in the Biological Assays described in Examples 13-14,
a PCR based strategy is employed to generate a GAS-SV40 promoter
sequence. The 5' primer contains four tandem copies of the GAS
binding site found in the IRF1 promoter and previously demonstrated
to bind STATs upon induction with a range of cytokines (Rothman et
al., Immunity 1:457-468 (1994).), although other GAS or ISRE
elements can be used instead. The 5' primer also contains 18bp of
sequence complementary to the SV40 early promoter sequence and is
flanked with an XhoI site. The sequence of the 5' primer is:
71 5':GCGCCTCGAGATTTCCCCGAAATCTAGATTTCC (SEQ ID NO: 3)
CCGAAATGATTTCCCCGAAATGATTTCCCCGAAATA TCTGCCATCTCAATTAG:3'
[1181] The downstream primer is complementary to the SV40 promoter
and is flanked with a Hind III site:
5':GCGGCAAGCTTTTTGCAAAGCCTAGGC:3' (SEQ ID NO:4)
[1182] PCR amplification is performed using the SV40 promoter
template present in the B-gal:promoter plasmid obtained from
Clontech. The resulting PCR fragment is digested with XhoI/Hind III
and subcloned into BLSK2-. (Stratagene.) Sequencing with forward
and reverse primers confirms that the insert contains the following
sequence:
72 5':CTCGAGATTTCCCCGAAATCTAGATTTCCCCGA (SEQ ID NO: 5)
AATGATTTCCCCGAAATGATTTCCCCGAAATATCTG
CCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCT
AACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTC
CGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTT
TATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGA
GCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGG CCTAGGCTTTTGCAAAAAGCTT:3'
[1183] With this GAS promoter element linked to the SV40 promoter,
a GAS:SEAP2 reporter construct is next engineered. Here, the
reporter molecule is a secreted alkaline phosphatase, or "SEAP."
Clearly, however, any reporter molecule can be instead of SEAP, in
this or in any of the other Examples. Well known reporter molecules
that can be used instead of SEAP include chloramphenicol
acetyltransferase (CAT), luciferase, alkaline phosphatase,
B-galactosidase, green fluorescent protein (GFP), or any protein
detectable by an antibody.
[1184] The above sequence confirmed synthetic GAS-SV40 promoter
element is subcloned into the pSEAP-Promoter vector obtained from
Clontech using HindIII and XhoI, effectively replacing the SV40
promoter with the amplified GAS:SV40 promoter element, to create
the GAS-SEAP vector. However, this vector does not contain a
neomycin resistance gene, and therefore, is not preferred for
mammalian expression systems.
[1185] Thus, in order to generate mammalian stable cell lines
expressing the GAS-SEAP reporter, the GAS-SEAP cassette is removed
from the GAS-SEAP vector using SalI and NotI, and inserted into a
backbone vector containing the neomycin resistance gene, such as
pGFP-1 (Clontech), using these restriction sites in the multiple
cloning site, to create the GAS-SEAP/Neo vector. Once this vector
is transfected into mammalian cells, this vector can then be used
as a reporter molecule for GAS binding as described in Examples
13-14.
[1186] Other constructs can be made using the above description and
replacing GAS with a different promoter sequence. For example,
construction of reporter molecules containing NFK-B and EGR
promoter sequences are described in Examples 15 and 16. However,
many other promoters can be substituted using the protocols
described in these Examples. For instance, SRE, IL-2, NFAT, or
Osteocalcin promoters can be substituted, alone or in combination
(e.g., GAS/NF-KB/EGR, GAS/NF-KB, I1-2/NFAT, or NF-KB/GAS).
Similarly, other cell lines can be used to test reporter construct
activity, such as HELA (epithelial), HUVEC (endothelial), Reh
(B-cell), Saos-2 (osteoblast), HUVAC (aortic), or
Cardiomyocyte.
Example 13
High-throughput Screening Assay for T-cell Activity
[1187] The following protocol is used to assess T-cell activity by
identifying factors, such as growth factors and cytokines, that may
proliferate or differentiate T-cells. T-cell activity is assessed
using the GAS/SEAP/Neo construct produced in Example 12. Thus,
factors that increase SEAP activity indicate the ability to
activate the Jaks-STATS signal transduction pathway. The T-cell
used in this assay is Jurkat T-cells (ATCC Accession No. TIB-152),
although Molt-3 cells (ATCC Accession No. CRL-1552) and Molt-4
cells (ATCC Accession No. CRL-1582) cells can also be used.
[1188] Jurkat T-cells are lymphoblastic CD4+ Th1 helper cells. In
order to generate stable cell lines, approximately 2 million Jurkat
cells are transfected with the GAS-SEAP/neo vector using DMRIE-C
(Life Technologies)(transfection procedure described below). The
transfected cells are seeded to a density of approximately 20,000
cells per well and transfectants resistant to 1 mg/ml genticin
selected. Resistant colonies are expanded and then tested for their
response to increasing concentrations of interferon gamma. The dose
response of a selected clone is demonstrated.
[1189] Specifically, the following protocol will yield sufficient
cells for 75 wells containing 200 ul of cells. Thus, it is either
scaled up, or performed in multiple to generate sufficient cells
for multiple 96 well plates. Jurkat cells are maintained in
RPMI+10% serum with 1%Pen-Strep. Combine 2.5 mls of OPTI-MEM (Life
Technologies) with 10 ug of plasmid DNA in a T25 flask. Add 2.5 ml
OPTI-MEM containing 50 ul of DMRIE-C and incubate at room
temperature for 1545 mins.
[1190] During the incubation period, count cell concentration, spin
down the required number of cells (10.sup.7 per transfection), and
resuspend in OPTI-MEM to a final concentration of 10.sup.7
cells/ml. Then add 1 ml of 1.times.10.sup.7 cells in OPTI-MEM to
T25 flask and incubate at 37.degree. C. for 6 hrs. After the
incubation, add 10 ml of RPMI+15% serum.
[1191] The Jurkat:GAS-SEAP stable reporter lines are maintained in
RPMI+10% serum, 1 mg/ml Genticin, and 1% Pen-Strep. These cells are
treated with supernatants containing a polypeptide as produced by
the protocol described in Example 11.
[1192] On the day of treatment with the supernatant, the cells
should be washed and resuspended in fresh RPMI+10% serum to a
density of 500,000 cells per ml. The exact number of cells required
will depend on the number of supernatants being screened. For one
96 well plate, approximately 10 million cells (for 10 plates, 100
million cells) are required.
[1193] Transfer the cells to a triangular reservoir boat, in order
to dispense the cells into a 96 well dish, using a 12 channel
pipette. Using a 12 channel pipette, transfer 200 ul of cells into
each well (therefore adding 100, 000 cells per well).
[1194] After all the plates have been seeded, 50 ul of the
supernatants are transferred directly from the 96 well plate
containing the supernatants into each well using a 12 channel
pipette. In addition, a dose of exogenous interferon gamma (0.1,
1.0, 10 ng) is added to wells H9, H10, and H11 to serve as
additional positive controls for the assay.
[1195] The 96 well dishes containing Jurkat cells treated with
supernatants are placed in an incubator for 48 hrs (note: this time
is variable between 48-72 hrs). 35 ul samples from each well are
then transferred to an opaque 96 well plate using a 12 channel
pipette. The opaque plates should be covered (using sellophene
covers) and stored at -20.degree. C. until SEAP assays are
performed according to Example 17. The plates containing the
remaining treated cells are placed at 4.degree. C. and serve as a
source of material for repeating the assay on a specific well if
desired.
[1196] As a positive control, 100 Unit/ml interferon gamma can be
used which is known to activate Jurkat T cells. Over 30 fold
induction is typically observed in the positive control wells.
[1197] The above protocol may be used in the generation of both
transient, as well as, stable transfected cells, which would be
apparent to those of skill in the art.
Example 14
High-throughput Screening Assay Identifying Myeloid Activity
[1198] The following protocol is used to assess myeloid activity by
identifying factors, such as growth factors and cytokines, that may
proliferate or differentiate myeloid cells. Myeloid cell activity
is assessed using the GAS/SEAP/Neo construct produced in Example
12. Thus, factors that increase SEAP activity indicate the ability
to activate the Jaks-STATS signal transduction pathway. The myeloid
cell used in this assay is U937, a pre-monocyte cell line, although
TF-1, HL60, or KG1 can be used.
[1199] To transiently transfect U937 cells with the GAS/SEAP/Neo
construct produced in Example 12, a DEAE-Dextran method (Kharbanda
et. al., 1994, Cell Growth & Differentiation, 5:259-265) is
used. First, harvest 2.times.10e.sup.7 U937 cells and wash with
PBS. The U937 cells are usually grown in RPMI 1640 medium
containing 10% heat-inactivated fetal bovine serum (FBS)
supplemented with 100 units/ml penicillin and 100 mg/ml
streptomycin.
[1200] Next, suspend the cells in 1 ml of 20 mM Tris-HCl (pH 7.4)
buffer containing 0.5 mg/ml DEAE-Dextran, 8 ug GAS-SEAP2 plasmid
DNA, 140 mM NaCl, 5 mM KCl, 375 uM Na.sub.2HPO.sub.4.7H.sub.2O, 1
mM MgCl.sub.2, and 675 uM CaCl.sub.2. Incubate at 37.degree. C. for
45 min.
[1201] Wash the cells with RPMI 1640 medium containing 10% FBS and
then resuspend in 10 ml complete medium and incubate at 37.degree.
C. for 36 hr.
[1202] The GAS-SEAP/U937 stable cells are obtained by growing the
cells in 400 ug/ml G418. The G418-free medium is used for routine
growth but every one to two months, the cells should be re-grown in
400 ug/ml G418 for couple of passages.
[1203] These cells are tested by harvesting 1.times.10.sup.8 cells
(this is enough for ten 96-well plates assay) and wash with PBS.
Suspend the cells in 200 ml above described growth medium, with a
final density of 5.times.10.sup.5 cells/ml. Plate 200 ul cells per
well in the 96-well plate (or 1.times.10.sup.5 cells/well).
[1204] Add 50 ul of the supernatant prepared by the protocol
described in Example 11. Incubate at 37.degree. C. for 48 to 72 hr.
As a positive control, 100 Unit/ml interferon gamma can be used
which is known to activate U937 cells. Over 30 fold induction is
typically observed in the positive control wells. SEAP assay the
supernatant according to the protocol described in Example 17.
Example 15
High-throughput Screening Assay Identifying Neuronal Activity
[1205] When cells undergo differentiation and proliferation, a
group of genes are activated through many different signal
transduction pathways. One of these genes, EGR1 (early growth
response gene 1), is induced in various tissues and cell types upon
activation. The promoter of EGR1 is responsible for such induction.
Using the EGR1 promoter linked to reporter molecules, activation of
cells can be assessed.
[1206] Particularly, the following protocol is used to assess
neuronal activity in PC12 cell lines. PC12 cells (rat
phenochromocytoma cells) are known to proliferate and/or
differentiate by activation with a number of mitogens, such as TPA
(tetradecanoyl phorbol acetate), NGF (nerve growth factor), and EGF
(epidermal growth factor). The EGRI gene expression is activated
during this treatment. Thus, by stably transfecting PC12 cells with
a construct containing an EGR promoter linked to SEAP reporter,
activation of PC12 cells can be assessed.
[1207] The EGR/SEAP reporter construct can be assembled by the
following protocol. The EGR-1 promoter sequence (-633 to +l)
(Sakamoto K et al., Oncogene 6:867-871 (1991)) can be PCR amplified
from human genomic DNA using the following primers:
73 (SEQ ID NO: 6) 5' GCGCTCGAGGGATGACAGCGATAGAACCCCGG-3' (SEQ ID
NO: 7) 5' GCGAAGCTTCGCGACTCCCCGGATCCGCCTC-3'
[1208] Using the GAS:SEAP/Neo vector produced in Example 12, EGR1
amplified product can then be inserted into this vector. Linearize
the GAS:SEAP/Neo vector using restriction enzymes XhoI/HindIII,
removing the GAS/SV40 stuffer. Restrict the EGRI amplified product
with these same enzymes. Ligate the vector and the EGR1
promoter.
[1209] To prepare 96 well-plates for cell culture, two mls of a
coating solution (1:30 dilution of collagen type I (Upstate Biotech
Inc. Cat#08-115) in 30% ethanol (filter sterilized)) is added per
one 10 cm plate or 50 ml per well of the 96-well plate, and allowed
to air dry for 2 hr.
[1210] PC12 cells are routinely grown in RPMI-1640 medium (Bio
Whittaker) containing 10% horse serum (JRH BIOSCIENCES, Cat. #
12449-78P), 5% heat-inactivated fetal bovine serum (FBS)
supplemented with 100 units/mil penicillin and 100 ug/ml
streptomycin on a precoated 10 cm tissue culture dish. One to four
split is done every three to four days. Cells are removed from the
plates by scraping and resuspended with pipetting up and down for
more than 15 times.
[1211] Transfect the EGR/SEAP/Neo construct into PC12 using the
Lipofectamine protocol described in Example 11. EGR-SEAP/PC 12
stable cells are obtained by growing the cells in 300 ug/ml G418.
The G418-free medium is used for routine growth but every one to
two months, the cells should be re-grown in 300 ug/ml G418 for
couple of passages.
[1212] To assay for neuronal activity, a 10 cm plate with cells
around 70 to 80% confluent is screened by removing the old medium.
Wash the cells once with PBS (Phosphate buffered saline). Then
starve the cells in low serum medium (RPMI-1640 containing 1% horse
serum and 0.5% FBS with antibiotics) overnight.
[1213] The next morning, remove the medium and wash the cells with
PBS. Scrape off the cells from the plate, suspend the cells well in
2 ml low serum medium. Count the cell number and add more low serum
medium to reach final cell density as 5.times.10.sup.5
cells/ml.
[1214] Add 200 ul of the cell suspension to each well of 96-well
plate (equivalent to 1.times.10.sup.5 cells/well). Add 50 ul
supernatant produced by Example 11, 37.degree. C. for 48 to 72 hr.
As a positive control, a growth factor known to activate PC12 cells
through EGR can be used, such as 50 ng/ul of Neuronal Growth Factor
(NGF). Over fifty-fold induction of SEAP is typically seen in the
positive control wells. SEAP assay the supernatant according to
Example 17.
Example 16
High-throughput Screening Assay for T-cell Activity
[1215] NF-.kappa.B (Nuclear Factor .kappa.B) is a transcription
factor activated by a wide variety of agents including the
inflammatory cytokines IL-1 and TNF, CD30 and CD40,
lymphotoxin-alpha and lymphotoxin-beta, by exposure to LPS or
thrombin, and by expression of certain viral gene products. As a
transcription factor, NF-.kappa.B regulates the expression of genes
involved in immune cell activation, control of apoptosis
(NF-.kappa.B appears to shield cells from apoptosis), B and T-cell
development, anti-viral and antimicrobial responses, and multiple
stress responses.
[1216] In non-stimulated conditions, NF-.kappa.B is retained in the
cytoplasm with I-KB (inhibitor KB). However, upon stimulation,
I-.kappa.B is phosphorylated and degraded, causing NF-.kappa.B to
shuttle to the nucleus, thereby activating transcription of target
genes. Target genes activated by NF-.kappa.B include IL-2, IL-6,
GM-CSF, ICAM-1 and class 1 MHC.
[1217] Due to its central role and ability to respond to a range of
stimuli, reporter constructs utilizing the NF-.kappa.B promoter
element are used to screen the supernatants produced in Example 11.
Activators or inhibitors of NF-KB would be useful in treating
diseases. For example, inhibitors of NF-.kappa.B could be used to
treat those diseases related to the acute or chronic activation of
NF-KB, such as rheumatoid arthritis.
[1218] To construct a vector containing the NF-.kappa.B promoter
element, a PCR based strategy is employed. The upstream primer
contains four tandem copies of the NF-.kappa.B binding site
(GGGGACTTTCCC) (SEQ ID NO:8), 18 bp of sequence complementary to
the 5' end of the SV40 early promoter sequence, and is flanked with
an XhoI site:
74 5':GCGGCCTCGAGGGGACTTTCCCGGGGACTTTCCG (SEQ ID NO:9)
GGGACTTTCCGGGACTTTCCATCCTGCCATCTCAATT AG:3'
[1219] The downstream primer is complementary to the 3' end of the
SV40 promoter and is flanked with a Hind III site:
75 5':GCGGCAAGCTTTTTGCAAAGCCTAGGC:3' (SEQ ID NO:4)
[1220] PCR amplification is performed using the SV40 promoter
template present in the pB-gal:promoter plasmid obtained from
Clontech. The resulting PCR fragment is digested with XhoI and Hind
III and subcloned into BLSK2-. (Stratagene) Sequencing with the T7
and T3 primers confirms the insert contains the following
sequence:
76 5':CTCGAGGGGACTTTCCCGGGGACTTTCCGGGGAC (SEQ ID NO:10)
TTTCCGGGACTTTCCATCTGCCATCTCAATTAGTCAG
CAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCC
CCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCAT
GGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGG
CCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGG
AGGCTTTTTTGGAGGCCTAGGCTTTTGCAAAAAGCT T:3'
[1221] Next, replace the SV40 minimal promoter element present in
the pSEAP2-promoter plasmid (Clontech) with this NF-.kappa.B/SV40
fragment using XhoI and HindIII. However, this vector does not
contain a neomycin resistance gene, and therefore, is not preferred
for mammalian expression systems.
[1222] In order to generate stable mammalian cell lines, the
NF-KB/SV40/SEAP cassette is removed from the above NF-.kappa.B/SEAP
vector using restriction enzymes SalI and NotI, and inserted into a
vector containing neomycin resistance. Particularly, the
NF-.kappa.B/SV40/SEAP cassette was inserted into pGFP-1 (Clontech),
replacing the GFP gene, after restricting pGFP-1 with SalI and
NotI.
[1223] Once NF-.kappa.B/SV40/SEAP/Neo vector is created, stable
Jurkat T-cells are created and maintained according to the protocol
described in Example 13. Similarly, the method for assaying
supernatants with these stable Jurkat T-cells is also described in
Example 13. As a positive control, exogenous TNF alpha (0.1, 1, 10
ng) is added to wells H9, H10, and H11, with a 5-10 fold activation
typically observed.
Example 17
Assay for SEAP Activity
[1224] As a reporter molecule for the assays described in Examples
13-16, SEAP activity is assayed using the Tropix Phospho-light Kit
(Cat. BP-400) according to the following general procedure. The
Tropix Phospho-light Kit supplies the Dilution, Assay, and Reaction
Buffers used below.
[1225] Prime a dispenser with the 2.5.times. Dilution Buffer and
dispense 15 .mu.l of 2.5.times. dilution buffer into Optiplates
containing 35 .mu.l of a supernatant. Seal the plates with a
plastic sealer and incubate at 65.degree. C. for 30 min. Separate
the Optiplates to avoid uneven heating.
[1226] Cool the samples to room temperature for 15 minutes. Empty
the dispenser and prime with the Assay Buffer. Add 50 .mu.l Assay
Buffer and incubate at room temperature 5 min. Empty the dispenser
and prime with the Reaction Buffer (see the table below). Add 50
.mu.l Reaction Buffer and incubate at room temperature for 20
minutes. Since the intensity of the chemiluminescent signal is time
dependent, and it takes about 10 minutes to read 5 plates on
luminometer, one should treat 5 plates at each time and start the
second set 10 minutes later.
[1227] Read the relative light unit in the luminometer. Set H12 as
blank, and print the results. An increase in chemiluminescence
indicates reporter activity.
77 Reaction Buffer Formulation: # of plates Rxn buffer diluent (ml)
CSPD (ml) 10 60 3 11 65 3.25 12 70 3.5 13 75 3.75 14 80 4 15 85
4.25 16 90 4.5 17 95 4.75 18 100 5 19 105 5.25 20 110 5.5 21 115
5.75 22 120 6 23 125 6.25 24 130 6.5 25 135 6.75 26 140 7 27 145
7.25 28 150 7.5 29 155 7.75 30 160 8 31 165 8.25 32 170 8.5 33 175
8.75 34 180 9 35 185 9.25 36 190 9.5 37 195 9.75 38 200 10 39 205
10.25 40 210 10.5 41 215 10.75 42 220 11 43 225 11.25 44 230 11.5
45 235 11.75 46 240 12 47 245 12.25 48 250 12.5 49 255 12.75 50 260
13
Example 18
High-throughput Screening Assay Identifying Changes in Small
Molecule Concentration and Membrane Permeability
[1228] Binding of a ligand to a receptor is known to alter
intracellular levels of small molecules, such as calcium,
potassium, sodium, and pH, as well as alter membrane potential.
These alterations can be measured in an assay to identify
supernatants which bind to receptors of a particular cell. Although
the following protocol describes an assay for calcium, this
protocol can easily be modified to detect changes in potassium,
sodium, pH, membrane potential, or any other small molecule which
is detectable by a fluorescent probe.
[1229] The following assay uses Fluorometric Imaging Plate Reader
("FLIPR") to measure changes in fluorescent molecules (Molecular
Probes) that bind small molecules. Clearly, any fluorescent
molecule detecting a small molecule can be used instead of the
calcium fluorescent molecule, fluo-4 (Molecular Probes, Inc.;
catalog no. F-14202),used here.
[1230] For adherent cells, seed the cells at 10,000 -20,000
cells/well in a Co-star black 96-well plate with clear bottom. The
plate is incubated in a CO.sub.2 incubator for 20 hours. The
adherent cells are washed two times in Biotek washer with 200 ul of
HBSS (Hank's Balanced Salt Solution) leaving 100 ul of buffer after
the final wash.
[1231] A stock solution of 1 mg/ml fluo-4 is made in 10% pluronic
acid DMSO. To load the cells with fluo-4, 50 ul of 12 ug/ml fluo-4
is added to each well. The plate is incubated at 37.degree. C. in a
CO.sub.2 incubator for 60 min. The plate is washed four times in
the Biotek washer with HBSS leaving 100 ul of buffer.
[1232] For non-adherent cells, the cells are spun down from culture
media. Cells are re-suspended to 2-5.times.10.sup.6 cells/ml with
HBSS in a 50-ml conical tube. 4 ul of 1 mg/ml fluo-4 solution in
10% pluronic acid DMSO is added to each ml of cell suspension. The
tube is then placed in a 37.degree. C. water bath for 30-60 min.
The cells are washed twice with HBSS, resuspended to
1.times.10.sup.6 cells/ml, and dispensed into a microplate, 100
ul/well. The plate is centrifuged at 1000 rpm for 5 min. The plate
is then washed once in Denley CellWash with 200 ul, followed by an
aspiration step to 100 ul final volume.
[1233] For a non-cell based assay, each well contains a fluorescent
molecule, such as fluo-4. The supernatant is added to the well, and
a change in fluorescence is detected.
[1234] To measure the fluorescence of intracellular calcium, the
FLIPR is set for the following parameters: (1) System gain is
300-800 mW; (2) Exposure time is 0.4 second; (3) Camera F/stop is
F/2; (4) Excitation is 488 nm; (5) Emission is 530 nm; and (6)
Sample addition is 50 ul. Increased emission at 530 nm indicates an
extracellular signaling event which has resulted in an increase in
the intracellular Ca.sup.++ concentration.
Example 19
High-throughput Screening Assay Identifying Tyrosine Kinase
Activity
[1235] The Protein Tyrosine Kinases (PTK) represent a diverse group
of transmembrane and cytoplasmic kinases. Within the Receptor
Protein Tyrosine Kinase RPTK) group are receptors for a range of
mitogenic and metabolic growth factors including the PDGF, FGF,
EGF, NGF, HGF and Insulin receptor subfamilies. In addition there
are a large family of RPTKs for which the corresponding ligand is
unknown. Ligands for RPTKs include mainly secreted small proteins,
but also membrane-bound and extracellular matrix proteins.
[1236] Activation of RPTK by ligands involves ligand-mediated
receptor dimerization, resulting in transphosphorylation of the
receptor subunits and activation of the cytoplasmic tyrosine
kinases. The cytoplasmic tyrosine kinases include receptor
associated tyrosine kinases of the src-family (e.g., src, yes, Ick,
lyn, fyn) and non-receptor linked and cytosolic protein tyrosine
kinases, such as the Jak family, members of which mediate signal
transduction triggered by the cytokine superfamily of receptors
(e.g., the Interleukins, Interferons, GM-CSF, and Leptin).
[1237] Because of the wide range of known factors capable of
stimulating tyrosine kinase activity, the identification of novel
human secreted proteins capable of activating tyrosine kinase
signal transduction pathways are of interest. Therefore, the
following protocol is designed to identify those novel human
secreted proteins capable of activating the tyrosine kinase signal
transduction pathways.
[1238] Seed target cells (e.g., primary keratinocytes) at a density
of approximately 25,000 cells per well in a 96 well Loprodyne
Silent Screen Plates purchased from Nalge Nunc (Naperville, Ill.).
The plates are sterilized with two 30 minute rinses with 100%
ethanol, rinsed with water and dried overnight. Some plates are
coated for 2 hr with 100 ml of cell culture grade type I collagen
(50 mg/ml), gelatin (2%) or polylysine (50 mg/ml), all of which can
be purchased from Sigma Chemicals (St. Louis, Mo.) or 10% Matrigel
purchased from Becton Dickinson (Bedford, Mass.), or calf serum,
rinsed with PBS and stored at 4.degree. C. Cell growth on these
plates is assayed by seeding 5,000 cells/well in growth medium and
indirect quantitation of cell number through use of alamarBlue as
described by the manufacturer Alamar Biosciences, Inc. (Sacramento,
Calif.) after 48 hr. Falcon plate covers #3071 from Becton
Dickinson (Bedford, Mass.) are used to cover the Loprodyne Silent
Screen Plates. Falcon Microtest III cell culture plates can also be
used in some proliferation experiments.
[1239] To prepare extracts, A431 cells are seeded onto the nylon
membranes of Loprodyne plates (20,000/200ml/well) and cultured
overnight in complete medium. Cells are quiesced by incubation in
serum-free basal medium for 24 hr. After 5-20 minutes treatment
with EGF (60 ng/ml) or 50 ul of the supernatant produced in Example
11, the medium was removed and 100 ml of extraction buffer ((20 mM
HEPES pH 7.5, 0.15 M NaCl, 1% Triton X-100, 0.1% SDS, 2 mM Na3VO4,
2 mM Na4P2O7 and a cocktail of protease inhibitors (# 1836170)
obtained from Boeheringer Mannheim (Indianapolis, Ind.) is added to
each well and the plate is shaken on a rotating shaker for 5
minutes at 4.degree. C. The plate is then placed in a vacuum
transfer manifold and the extract filtered through the 0.45 mm
membrane bottoms of each well using house vacuum. Extracts are
collected in a 96-well catch/assay plate in the bottom of the
vacuum manifold and immediately placed on ice. To obtain extracts
clarified by centrifugation, the content of each well, after
detergent solubilization for 5 minutes, is removed and centrifuged
for 15 minutes at 4.degree. C. at 16,000.times. g.
[1240] Test the filtered extracts for levels of tyrosine kinase
activity. Although many methods of detecting tyrosine kinase
activity are known, one method is described here.
[1241] Generally, the tyrosine kinase activity of a supernatant is
evaluated by determining its ability to phosphorylate a tyrosine
residue on a specific substrate (a biotinylated peptide).
Biotinylated peptides that can be used for this purpose include
PSK1 (corresponding to amino acids 6-20 of the cell division kinase
cdc2-p34) and PSK2 (corresponding to amino acids 1-17 of gastrin).
Both peptides are substrates for a range of tyrosine kinases and
are available from Boehringer Mannheim.
[1242] The tyrosine kinase reaction is set up by adding the
following components in order. First, add 10 ul of 5 uM
Biotinylated Peptide, then 10 ul ATP/Mg.sub.2+ (5 mM ATP/50 mM
MgCl.sub.2), then 10 ul of 5.times. Assay Buffer (40 mM imidazole
hydrochloride, pH 7.3, 40 mM beta-glycerophosphate, 1 mM EGTA, 100
mM MgCl.sub.2, 5 mM MnCl.sub.2, 0.5 mg/ml BSA), then 5 ul of Sodium
Vanadate (1 mM), and then 5 ul of water. Mix the components gently
and preincubate the reaction mix at 30.degree. C. for 2 min.
Initial the reaction by adding 10 ul of the control enzyme or the
filtered supernatant.
[1243] The tyrosine kinase assay reaction is then terminated by
adding 10 ul of 120 mm EDTA and place the reactions on ice.
[1244] Tyrosine kinase activity is determined by transferring 50 ul
aliquot of reaction mixture to a microtiter plate (MTP) module and
incubating at 37.degree. C. for 20 min. This allows the
streptavadin coated 96 well plate to associate with the
biotinylated peptide. Wash the MTP module with 300 ul/well of PBS
four times. Next add 75 ul of anti-phospotyrosine antibody
conjugated to horse radish peroxidase(anti-P-Tyr-POD(0.5u/ml)) to
each well and incubate at 37.degree. C. for one hour. Wash the well
as above.
[1245] Next add 100 ul of peroxidase substrate solution (Boehringer
Mannheim) and incubate at room temperature for at least 5 mins (up
to 30 min). Measure the absorbance of the sample at 405 nm by using
ELISA reader. The level of bound peroxidase activity is quantitated
using an ELISA reader and reflects the level of tyrosine kinase
activity.
Example 20
High-throughput Screening Assay Identifying Phosphorylation
Activity
[1246] As a potential alternative and/or compliment to the assay of
protein tyrosine kinase activity described in Example 19, an assay
which detects activation (phosphorylation) of major intracellular
signal transduction intermediates can also be used. For example, as
described below one particular assay can detect tyrosine
phosphorylation of the Erk-1 and Erk-2 kinases. However,
phosphorylation of other molecules, such as Raf, JNK, p38 MAP, Map
kinase kinase (MEK), MEK kinase, Src, Muscle specific kinase
(MuSK), IRAK, Tec, and Janus, as well as any other phosphoserine,
phosphotyrosine, or phosphothreonine molecule, can be detected by
substituting these molecules for Erk-1 or Erk-2 in the following
assay.
[1247] Specifically, assay plates are made by coating the wells of
a 96-well ELISA plate with 0.1 ml of protein G (1 ug/ml) for 2 hr
at room temp, (RT). The plates are then rinsed with PBS and blocked
with 3% BSA/PBS for 1 hr at RT. The protein G plates are then
treated with 2 commercial monoclonal antibodies (100 ng/well)
against Erk-1 and Erk-2 (1 hr at RT) (Santa Cruz Biotechnology).
(To detect other molecules, this step can easily be modified by
substituting a monoclonal antibody detecting any of the above
described molecules.) After 3-5 rinses with PBS, the plates are
stored at 4.degree. C. until use.
[1248] A431 cells are seeded at 20,000/well in a 96-well Loprodyne
filterplate and cultured overnight in growth medium. The cells are
then starved for 48 hr in basal medium (DMEM) and then treated with
EGF (6ng/well) or 50 ul of the supernatants obtained in Example 11
for 5-20 minutes. The cells are then solubilized and extracts
filtered directly into the assay plate.
[1249] After incubation with the extract for 1 hr at RT, the wells
are again rinsed. As a positive control, a commercial preparation
of MAP kinase (10 ng/well) is used in place of A43 1 extract.
Plates are then treated with a commercial polyclonal (rabbit)
antibody (1 ug/mil) which specifically recognizes the
phosphorylated epitope of the Erk-1 and Erk-2 kinases (1 hr at RT).
This antibody is biotinylated by standard procedures. The bound
polyclonal antibody is then quantitated by successive incubations
with Europium-streptavidin and Europium fluorescence enhancing
reagent in the Wallac DELFIA instrument (time-resolved
fluorescence). An increased fluorescent signal over background
indicates a phosphorylation.
Example 21
Method of Determining Alterations in a Gene Corresponding to a
Polynucleotide
[1250] RNA isolated from entire families or individual patients
presenting with a phenotype of interest (such as a disease) is be
isolated. cDNA is then generated from these RNA samples using
protocols known in the art. (See, Sambrook.) The cDNA is then used
as a template for PCR, employing primers surrounding regions of
interest in SEQ ID NO:X. Suggested PCR conditions consist of 35
cycles at 95.degree. C. for 30 seconds; 60-120 seconds at
52-58.degree. C.; and 60-120 seconds at 70.degree. C., using buffer
solutions described in Sidransky, D., et al., Science 252:706
(1991).
[1251] PCR products are then sequenced using primers labeled at
their 5' end with T4 polynucleotide kinase, employing SequiTherm
Polymerase. (Epicentre Technologies). The intron-exon borders of
selected exons is also determined and genomic PCR products analyzed
to confirm the results. PCR products harboring suspected mutations
is then cloned and sequenced to validate the results of the direct
sequencing.
[1252] PCR products is cloned into T-tailed vectors as described in
Holton, T. A. and Graham, M. W., Nucleic Acids Research, 19:1156
(1991) and sequenced with T7 polymerase (United States
Biochemical). Affected individuals are identified by mutations not
present in unaffected individuals.
[1253] Genomic rearrangements are also observed as a method of
determining alterations in a gene corresponding to a
polynucleotide. Genomic clones isolated according to Example 2 are
nick-translated with digoxigenindeoxy-uridine 5'-triphosphate
(Boehringer Manheim), and FISH performed as described in Johnson,
Cg. et al., Methods Cell Biol. 35:73-99 (1991). Hybridization with
the labeled probe is carried out using a vast excess of human cot-1
DNA for specific hybridization to the corresponding genomic
locus.
[1254] Chromosomes are counterstained with
4,6-diamino-2-phenylidole and propidium iodide, producing a
combination of C- and R-bands. Aligned images for precise mapping
are obtained using a triple-band filter set (Chroma Technology,
Brattleboro, Vt.) in combination with a cooled charge-coupled
device camera (Photometrics, Tucson, Ariz.) and variable excitation
wavelength filters. (Johnson, Cv. et al., Genet. Anal. Tech. Appl.,
8:75 (1991).) Image collection, analysis and chromosomal fractional
length measurements are performed using the ISee Graphical Program
System. (Inovision Corporation, Durham, N.C.) Chromosome
alterations of the genomic region hybridized by the probe are
identified as insertions, deletions, and translocations. These
alterations are used as a diagnostic marker for an associated
disease.
Example 22
Method of Detecting Abnormal Levels of a Polypeptide in a
Biological Sample
[1255] A polypeptide of the present invention can be detected in a
biological sample, and if an increased or decreased level of the
polypeptide is detected, this polypeptide is a marker for a
particular phenotype. Methods of detection are numerous, and thus,
it is understood that one skilled in the art can modify the
following assay to fit their particular needs.
[1256] For example, antibody-sandwich ELISAs are used to detect
polypeptides in a sample, preferably a biological sample. Wells of
a microtiter plate are coated with specific antibodies, at a final
concentration of 0.2 to 10 ug/ml. The antibodies are either
monoclonal or polyclonal and are produced by the method described
in Example 10. The wells are blocked so that non-specific binding
of the polypeptide to the well is reduced.
[1257] The coated wells are then incubated for >2 hours at RT
with a sample containing the polypeptide. Preferably, serial
dilutions of the sample should be used to validate results. The
plates are then washed three times with deionized or distilled
water to remove unbounded polypeptide.
[1258] Next, 50 ul of specific antibody-alkaline phosphatase
conjugate, at a concentration of 25-400 ng, is added and incubated
for 2 hours at room temperature. The plates are again washed three
times with deionized or distilled water to remove unbounded
conjugate.
[1259] Add 75 ul of 4-methylumbelliferyl phosphate (MUP) or
p-nitrophenyl phosphate (NPP) substrate solution to each well and
incubate 1 hour at room temperature. Measure the reaction by a
microtiter plate reader. Prepare a standard curve, using serial
dilutions of a control sample, and plot polypeptide concentration
on the X-axis (log scale) and fluorescence or absorbance of the
Y-axis (linear scale). Interpolate the concentration of the
polypeptide in the sample using the standard curve.
Example 23
Formulating a Polypeptide
[1260] The secreted polypeptide composition will be formulated and
dosed in a fashion consistent with good medical practice, taking
into account the clinical condition of the individual patient
(especially the side effects of treatment with the secreted
polypeptide alone), the site of delivery, the method of
administration, the scheduling of administration, and other factors
known to practitioners. The "effective amount" for purposes herein
is thus determined by such considerations.
[1261] As a general proposition, the total pharmaceutically
effective amount of secreted polypeptide administered parenterally
per dose will be in the range of about 1 .mu.g/kg/day to 10
mg/kg/day of patient body weight, although, as noted above, this
will be subject to therapeutic discretion. More preferably, this
dose is at least 0.01 mg/kg/day, and most preferably for humans
between about 0.01 and 1 mg/kg/day for the hormone. If given
continuously, the secreted polypeptide is typically administered at
a dose rate of about 1 .mu.g/kg/hour to about 50 .mu.g/kg/hour,
either by 1-4 injections per day or by continuous subcutaneous
infusions, for example, using a mini-pump. An intravenous bag
solution may also be employed. The length of treatment needed to
observe changes and the interval following treatment for responses
to occur appears to vary depending on the desired effect.
[1262] Pharmaceutical compositions containing the secreted protein
of the invention are administered orally, rectally, parenterally,
intracistemally, intravaginally, intraperitoneally, topically (as
by powders, ointments, gels, drops or transdermal patch), bucally,
or as an oral or nasal spray. "Pharmaceutically acceptable carrier"
refers to a non-toxic solid, semisolid or liquid filler, diluent,
encapsulating material or formulation auxiliary of any type. The
term "parenteral" as used herein refers to modes of administration
which include intravenous, intramuscular, intraperitoneal,
intrasternal, subcutaneous and intraarticular injection and
infusion.
[1263] The secreted polypeptide is also suitably administered by
sustained-release systems. Suitable examples of sustained-release
compositions include semi-permeable polymer matrices in the form of
shaped articles, e.g., films, or mirocapsules. Sustained-release
matrices include polylactides (U.S. Pat. No. 3,773,919, EP 58,481),
copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman,
U. et al., Biopolymers 22:547-556 (1983)), poly (2-hydroxyethyl
methacrylate) (R. Langer et al., J. Biomed. Mater. Res. 15:167-277
(1981), and R. Langer, Chem. Tech. 12:98-105 (1982)), ethylene
vinyl acetate (R. Langer et al.) or poly-D-(-)-3-hydroxybutyric
acid (EP 133,988). Sustained-release compositions also include
liposomally entrapped polypeptides. Liposomes containing the
secreted polypeptide are prepared by methods known per se: DE
3,218,121; Epstein et al., Proc. Natl. Acad. Sci. USA 82:3688-3692
(1985); Hwang et al., Proc. Natl. Acad. Sci. USA 77:4030-4034
(1980); EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP 142,641;
Japanese Pat. Appl. 83-118008; U.S. Pat. Nos. 4,485,045 and
4,544,545; and EP 102,324. Ordinarily, the liposomes are of the
small (about 200-800 Angstroms) unilamellar type in which the lipid
content is greater than about 30 mol. percent cholesterol, the
selected proportion being adjusted for the optimal secreted
polypeptide therapy.
[1264] For parenteral administration, in one embodiment, the
secreted polypeptide is formulated generally by mixing it at the
desired degree of purity, in a unit dosage injectable form
(solution, suspension, or emulsion), with a pharmaceutically
acceptable carrier, i.e., one that is non-toxic to recipients at
the dosages and concentrations employed and is compatible with
other ingredients of the formulation. For example, the formulation
preferably does not include oxidizing agents and other compounds
that are known to be deleterious to polypeptides.
[1265] Generally, the formulations are prepared by contacting the
polypeptide uniformly and intimately with liquid carriers or finely
divided solid carriers or both. Then, if necessary, the product is
shaped into the desired formulation. Preferably the carrier is a
parenteral carrier, more preferably a solution that is isotonic
with the blood of the recipient. Examples of such carrier vehicles
include water, saline, Ringer's solution, and dextrose solution.
Non-aqueous vehicles such as fixed oils and ethyl oleate are also
useful herein, as well as liposomes.
[1266] The carrier suitably contains minor amounts of additives
such as substances that enhance isotonicity and chemical stability.
Such materials are non-toxic to recipients at the dosages and
concentrations employed, and include buffers such as phosphate,
citrate, succinate, acetic acid, and other organic acids or their
salts; antioxidants such as ascorbic acid; low molecular weight
(less than about ten residues) polypeptides, e.g., polyarginine or
tripeptides; proteins, such as serum albumin, gelatin, or
immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone;
amino acids, such as glycine, glutamic acid, aspartic acid, or
arginine; monosaccharides, disaccharides, and other carbohydrates
including cellulose or its derivatives, glucose, manose, or
dextrins; chelating agents such as EDTA; sugar alcohols such as
mannitol or sorbitol; counterions such as sodium; and/or nonionic
surfactants such as polysorbates, poloxamers, or PEG.
[1267] The secreted polypeptide is typically formulated in such
vehicles at a concentration of about 0.1 mg/ml to 100 mg/ml,
preferably 1-10 mg/mil, at a pH of about 3 to 8. It will be
understood that the use of certain of the foregoing excipients,
carriers, or stabilizers will result in the formation of
polypeptide salts.
[1268] Any polypeptide to be used for therapeutic administration
can be sterile. Sterility is readily accomplished by filtration
through sterile filtration membranes (e.g., 0.2 micron membranes).
Therapeutic polypeptide compositions generally are placed into a
container having a sterile access port, for example, an intravenous
solution bag or vial having a stopper pierceable by a hypodermic
injection needle.
[1269] Polypeptides ordinarily will be stored in unit or multi-dose
containers, for example, sealed ampoules or vials; as an aqueous
solution or as a lyophilized formulation for reconstitution. As an
example of a lyophilized formulation, 10-ml vials are filled with 5
ml of sterile-filtered 1% (w/v) aqueous polypeptide solution, and
the resulting mixture is lyophilized. The infusion solution is
prepared by reconstituting the lyophilized polypeptide using
bacteriostatic Water-for-Injection.
[1270] The invention also provides a pharmaceutical pack or kit
comprising one or more containers filled with one or more of the
ingredients of the pharmaceutical compositions of the invention.
Associated with such container(s) can be a notice in the form
prescribed by a governmental agency regulating the manufacture, use
or sale of pharmaceuticals or biological products, which notice
reflects approval by the agency of manufacture, use or sale for
human administration. In addition, the polypeptides of the present
invention may be employed in conjunction with other therapeutic
compounds.
Example 24
Method of Treating Decreased Levels of the Polypeptide
[1271] It will be appreciated that conditions caused by a decrease
in the standard or normal expression level of a secreted protein in
an individual can be treated by administering the polypeptide of
the present invention, preferably in the secreted form. Thus, the
invention also provides a method of treatment of an individual in
need of an increased level of the polypeptide comprising
administering to such an individual a pharmaceutical composition
comprising an amount of the polypeptide to increase the activity
level of the polypeptide in such an individual.
[1272] For example, a patient with decreased levels of a
polypeptide receives a daily dose 0.1-100 ug/kg of the polypeptide
for six consecutive days. Preferably, the polypeptide is in the
secreted form. The exact details of the dosing scheme, based on
administration and formulation, are provided in Example 23.
Example 25
Method of Treating Increased Levels of the Polypeptide
[1273] Antisense technology is used to inhibit production of a
polypeptide of the present invention. This technology is one
example of a method of decreasing levels of a polypeptide,
preferably a secreted form, due to a variety of etiologies, such as
cancer.
[1274] For example, a patient diagnosed with abnormally increased
levels of a polypeptide is administered intravenously antisense
polynucleotides at 0.5, 1.0, 1.5, 2.0 and 3.0 mg/kg day for 21
days. This treatment is repeated after a 7-day rest period if the
treatment was well tolerated. The formulation of the antisense
polynucleotide is provided in Example 23.
Example 26
Method of Treatment Using Gene Therapy
[1275] One method of gene therapy transplants fibroblasts, which
are capable of expressing a polypeptide, onto a patient. Generally,
fibroblasts are obtained from a subject by skin biopsy. The
resulting tissue is placed in tissue-culture medium and separated
into small pieces. Small chunks of the tissue are placed on a wet
surface of a tissue culture flask, approximately ten pieces are
placed in each flask. The flask is turned upside down, closed tight
and left at room temperature over night. After 24 hours at room
temperature, the flask is inverted and the chunks of tissue remain
fixed to the bottom of the flask and fresh media (e.g., Ham's F12
media, with 10% FBS, penicillin and streptomycin) is added. The
flasks are then incubated at 37.degree. C. for approximately one
week.
[1276] At this time, fresh media is added and subsequently changed
every several days. After an additional two weeks in culture, a
monolayer of fibroblasts emerge. The monolayer is trypsinized and
scaled into larger flasks.
[1277] pMV-7 (Kirschmeier, P. T. et al., DNA, 7:219-25 (1988)),
flanked by the long terminal repeats of the Moloney murine sarcoma
virus, is digested with EcoRI and HindIII and subsequently treated
with calf intestinal phosphatase. The linear vector is fractionated
on agarose gel and purified, using glass beads.
[1278] The cDNA encoding a polypeptide of the present invention can
be amplified using PCR primers which correspond to the 5' and 3'
end sequences respectively as set forth in Example 1. Preferably,
the 5' primer contains an EcoRI site and the 3' primer includes a
HindIII site. Equal quantities of the Moloney murine sarcoma virus
linear backbone and the amplified EcoRI and HindIII fragment are
added together, in the presence of T4 DNA ligase. The resulting
mixture is maintained under conditions appropriate for ligation of
the two fragments. The ligation mixture is then used to transform
bacteria HB 101, which are then plated onto agar containing
kanamycin for the purpose of confirming that the vector has the
gene of interest properly inserted.
[1279] The amphotropic pA317 or GP+am12 packaging cells are grown
in tissue culture to confluent density in Dulbecco's Modified
Eagles Medium (DMEM) with 10% calf serum (CS), penicillin and
streptomycin. The MSV vector containing the gene is then added to
the media and the packaging cells transduced with the vector. The
packaging cells now produce infectious viral particles containing
the gene (the packaging cells are now referred to as producer
cells).
[1280] Fresh media is added to the transduced producer cells, and
subsequently, the media is harvested from a 10 cm plate of
confluent producer cells. The spent media, containing the
infectious viral particles, is filtered through a millipore filter
to remove detached producer cells and this media is then used to
infect fibroblast cells. Media is removed from a sub-confluent
plate of fibroblasts and quickly replaced with the media from the
producer cells. This media is removed and replaced with fresh
media. If the titer of virus is high, then virtually all
fibroblasts will be infected and no selection is required. If the
titer is very low, then it is necessary to use a retroviral vector
that has a selectable marker, such as neo or his. Once the
fibroblasts have been efficiently infected, the fibroblasts are
analyzed to determine whether protein is produced.
[1281] The engineered fibroblasts are then transplanted onto the
host, either alone or after having been grown to confluence on
cytodex 3 microcarrier beads.
[1282] It will be clear that the invention may be practiced
otherwise than as particularly described in the foregoing
description and examples. Numerous modifications and variations of
the present invention are possible in light of the above teachings
and, therefore, are within the scope of the appended claims.
[1283] The entire disclosure of each document cited (including
patents, patent applications, journal articles, abstracts,
laboratory manuals, books, or other disclosures) in the Background
of the Invention, Detailed Description, and Examples is hereby
incorporated herein by reference. Further, the hard copy of the
sequence listing submitted herewith and the corresponding computer
readable form are both incorporated herein by reference in their
entireties.
Sequence CWU 0
0
* * * * *