U.S. patent application number 14/989741 was filed with the patent office on 2016-07-14 for peptide vaccines for cancers expressing tumor-associated antigens.
The applicant listed for this patent is OncoTherapy Science, Inc.. Invention is credited to Ryuji Ohsawa, Takuya Tsunoda.
Application Number | 20160200764 14/989741 |
Document ID | / |
Family ID | 41707002 |
Filed Date | 2016-07-14 |
United States Patent
Application |
20160200764 |
Kind Code |
A1 |
Tsunoda; Takuya ; et
al. |
July 14, 2016 |
PEPTIDE VACCINES FOR CANCERS EXPRESSING TUMOR-ASSOCIATED
ANTIGENS
Abstract
The present invention provides peptides having an amino acid
sequence as set forth in SEQ ID NO: 19, 22, 30, 34, 344, 358, 41,
44, 46, 48, 78, 376, 379, 80, 100, 101, 110, 111, 387, 112, 394,
114, 116, 117, 121, 395, 133, 135, 137, 426, 143, 147, 148, 149,
150, 152, 153, 154, 156, 160, 161, 162, 163, 166, 174, 178, 186,
194, 196, 202, 210, 213, 214, 217, 223, 227, 228, 233, 254, 271,
272 or 288, as well as peptides having the above-mentioned amino
acid sequences in which 1, 2, or several (e.g., up to 5) amino
acids are substituted, deleted, or added, provided the peptides
possess cytotoxic T cell inducibility. The present invention also
provides drugs for treating or preventing a disease associated with
over-expression of the CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A,
KNTC2, TTK and/or URLC10, e.g. cancers containing as an active
ingredient one or more of these peptides. The peptides of the
present invention find further utility as vaccines.
Inventors: |
Tsunoda; Takuya; (Kanagawa,
JP) ; Ohsawa; Ryuji; (Kanagawa, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
OncoTherapy Science, Inc. |
Kanagawa |
|
JP |
|
|
Family ID: |
41707002 |
Appl. No.: |
14/989741 |
Filed: |
January 6, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14274373 |
May 9, 2014 |
9284349 |
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14989741 |
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13744354 |
Jan 17, 2013 |
8759481 |
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14274373 |
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12542638 |
Aug 17, 2009 |
8383590 |
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13744354 |
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PCT/JP2008/000290 |
Feb 21, 2008 |
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12542638 |
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60902949 |
Feb 21, 2007 |
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61089973 |
Aug 19, 2008 |
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Current U.S.
Class: |
424/185.1 ;
435/375; 435/455; 530/300; 530/328; 536/23.5 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 2039/572 20130101; A61P 37/04 20180101; A61K 39/001122
20180801; A61K 38/00 20130101; C07K 4/12 20130101; G01N 2500/00
20130101; A61K 31/7088 20130101; A61K 39/0011 20130101; C07K 14/47
20130101; C07K 7/06 20130101; A61K 39/00 20130101 |
International
Class: |
C07K 7/06 20060101
C07K007/06; A61K 39/00 20060101 A61K039/00 |
Claims
1. An isolated peptide of (a) or (b) below: (a) an isolated peptide
of less than about 15 amino acids having cytotoxic T cell
inducibility, said peptide comprising the amino acid sequence of
SEQ ID NO: 80, 100 or 101; (b) an isolated peptide of less than
about 15 amino acids having cytotoxic T cell inducibility, wherein
said peptide comprises an amino acid sequence of SEQ ID NO: 80, 100
or 101 in which 1, 2, or several amino acids are substituted,
deleted, or added.
2. The peptide of claim 1, wherein the peptide is selected from the
group consisting of: (a) an isolated peptide consisting of the
amino acid sequence of SEQ ID NO: 80, 100 or 101; and (b) an
isolated peptide having cytotoxic T cell inducibility, wherein said
peptide consists of an amino acid sequence of SEQ ID NO: 80, 100 or
101 in which 1, 2, or several amino acids are substituted, deleted,
or added.
3. The peptide of claim 1, wherein the second amino acid from the
N-terminus of the amino acid sequence of SEQ ID NO: 80, 100 or 101
is substituted with phenylalanine, tyrosine, methionine, or
tryptophan.
4. The peptide of claim 1, wherein the C-terminal amino acid of the
amino acid sequence of SEQ ID NO: 80, 100 or 101 is substituted
with phenylalanine, leucine, isoleucine, tryptophan, or
methionine.
5. The peptide of claim 3, wherein the C-terminal amino acid of the
amino acid sequence of SEQ ID NO: 80, 100 or 101 is substituted
with phenylalanine, leucine, isoleucine, tryptophan, or
methionine.
6. A pharmaceutical composition for treating or preventing a
disease associated with over-expression of the gene of SEQ ID NO:
5, said composition comprising a peptide of claim 1 or a
polynucleotide encoding the peptide.
7. A pharmaceutical composition of claim 6, wherein the disease
associated with the gene of SEQ ID NO: 5 is cancer.
8. A pharmaceutical composition of claim 7, wherein the cancer is
selected from the group consisting of bladder cancer, breast
cancer, cervical cancer, cholangiocellular carcinoma, CML,
colorectal cancer, endometriosis, esophageal cancer, gastric
cancer, diffused type gastric cancer, liver cancer, NSCLC,
lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate
cancer, renal carcinoma, SCLC, soft tissue tumor and testicular
tumor.
9. A method of inducing antigen-presenting cells having cytotoxic T
cell inducibility, said method comprising the step of contacting an
antigen-presenting cell with a peptide of claim 1.
10. A method of inducing cytotoxic T cells by contacting a T cell
with a peptide of claim 1.
11. A method of inducing antigen-presenting cells having cytotoxic
T cell inducibility, said method comprising the step of
transferring a gene comprising a polynucleotide encoding a peptide
of claim 1 to an antigen-presenting cell.
12. A method of inducing a cytotoxic T cell, said method comprising
the steps of: (a) contacting an antigen-presenting cell with a
peptide of claim 1, and (b) mixing the antigen-presenting cells of
step (a) with a CD8+ T cell and co-culturing.
13. A vaccine for inhibiting proliferation of a cell expressing the
gene of SEQ ID NO: 5, wherein the vaccine comprises a peptide of
claim 1 as an active ingredient.
14. The vaccine of claim 13, wherein the cell expressing the gene
of SEQ ID NO: 5 is a cancer cell.
15. The vaccine of claim 14, wherein the cancer is selected from
the group consisting of bladder cancer, breast cancer, cervical
cancer, cholangiocellular carcinoma, CML, colorectal cancer,
endometriosis, esophageal cancer, gastric cancer, diffused type
gastric cancer, liver cancer, NSCLC, lymphoma, osteosarcoma,
ovarian cancer, pancreatic cancer, prostate cancer, renal
carcinoma, SCLC, soft tissue tumor and testicular tumor.
16. The vaccine of claim 15, formulated for administration to a
subject whose HLA antigen is HLA-A24.
17. A composition comprising a peptide of claim 1 or a
polynucleotide encoding the peptide.
18. A method of treating or preventing a disease associated with
over-expression of the gene of SEQ ID NO: 5 in a subject, said
method comprising administering to said subject a vaccine
comprising a peptide of claim 1, or a polynucleotide encoding said
peptide.
19. The method of claim 18, wherein the disease associated with
over-expression of the gene of SEQ ID NO: 5 is cancer.
20. The method of claim 19, wherein the cancer is selected from the
group consisting of bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, CML, colorectal cancer, endometriosis,
esophageal cancer, gastric cancer, diffused type gastric cancer,
liver cancer, NSCLC, lymphoma, osteosarcoma, ovarian cancer,
pancreatic cancer, prostate cancer, renal carcinoma, SCLC, soft
tissue tumor and testicular tumor.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is a division of U.S. application Ser. No.
14/274,373, filed May 9, 2014, which is a division of U.S.
application Ser. No. 13/744,354, filed Jan. 17, 2013, now U.S. Pat.
No. 8,759,481, which is a division of U.S. application Ser. No.
12/542,638, filed Aug. 17, 2009, now U.S. Pat. No. 8,383,590, which
is a continuation-in-part of PCT/JP2008/000290 (WO 2008/102557)
filed Feb. 21, 2008, which claims priority to U.S. Provisional
Application Ser. No. 60/902,949, filed Feb. 21, 2007. U.S.
application Ser. No. 12/542,638, filed Aug. 17, 2009, also claims
the benefit of priority to U.S. Provisional Application Ser. No.
61/089,973 filed Aug. 19, 2008. All of these applications are
incorporated herein by reference in their entirety.
REFERENCE TO A "SEQUENCE LISTING," A TABLE, OR A COMPUTER PROGRAM
LISTING APPENDIX SUBMITTED AS AN ASCII TEXT FILE
[0002] The Sequence Listing written in file
087331-0965171-020812US-SEQLIST.TXT, created on Nov. 21, 2015,
246,041 bytes, machine format IBM-PC, MS-Windows operating system,
is hereby incorporated by reference in its entirety for all
purposes.
TECHNICAL FIELD
[0003] The present invention relates to the field of biological
science, more specifically to the field of cancer therapy. In
particular, the present invention relates to novel immunogenic
peptides that serve as extremely effective as cancer vaccines, and
drugs for treating and preventing tumors containing such
peptides.
BACKGROUND ART
[0004] It has been demonstrated that CD8.sup.+ cytotoxic T
lymphocytes (CTLs) recognize epitope peptides derived from
tumor-associated antigens (TAAs) presented on MHC class I
molecules, and subsequently lyse the tumor cells. Since the
discovery of the MAGE family as the first example of TAAs, many
other TAAs have been discovered using immunological approaches
(Boon T. (1993) Int J Cancer 54: 177-80.; Boon T. et al., (1996) J
Exp Med 183: 725-9.; van der Bruggen P et al., (1991) Science 254:
1643-7.; Brichard V et al., (1993) J Exp Med 178: 489-95.; Kawakami
Y et al., (1994) J Exp Med 180: 347-52.). Some of them are now in
clinical development as targets of immunotherapy. TAAs discovered
to date include MAGE (van der Bruggen P et al., (1991) Science 254:
1643-7.), gp100 (Kawakami Y et al., (1994) J Exp Med 180: 347-52.),
SART (Shichijo S et al., (1998) J Exp Med 187:277-88.), and
NY-ESO-1 (Chen Y. T. et al., (1997) Proc. Natl. Acd. Sci. USA, 94:
1914-8.). On the other hand, certain gene products demonstrated to
be somewhat specifically over-expressed in tumor cells have been
shown to be recognized as targets for inducing cellular immune
responses. Such gene products include p53 (Umano Y et al., (2001)
Br J Cancer, 84:1052-7.), HER2/neu (Tanaka H et al., (2001) Br J
Cancer, 84: 94-9.), CEA (Nukaya I et al., (1999) Int. J. Cancer 80,
92-7.) and the like.
[0005] Despite significant progress in basic and clinical research
concerning TAAs (Rosenberg S A et al., (1998) Nature Med, 4:
321-7.; Mukherji B. et al., (1995) Proc Natl Acad Sci USA, 92:
8078-82.: Hu X et al., (1996) Cancer Res, 56: 2479-83.), only a
very limited number of candidate TAAs suitable for treatment of
cancers are presently available. TAAs that are abundantly expressed
in cancer cells, and whose expression is restricted to cancer
cells, would be promising candidates as immunotherapeutic
targets.
[0006] Both HLA-A24 and HLA-A0201 are common HLA alleles in the
Japanese and Caucasian populations (Date Y et. al., (1996) Tissue
Antigens 47: 93-101.; Kondo A et al., (1995) J Immunol 155:
4307-12.; Kubo R T et al., (1994) J Immunol 152: 3913-24.; Imanishi
et al., Proceeding of the eleventh International Histocompatibility
Workshop and Conference Oxford University Press, Oxford, 1065
(1992); Williams F et al., (1997) Tissue Antigen 49: 129-33.).
Thus, antigenic peptides of cancers presented by these HLA alleles
may find particular utility in the treatment of cancers among
Japanese and Caucasian patients. Further, it is known that the
induction of low-affinity CTL in vitro usually results from
exposure to high concentrations of peptides, generating a high
level of specific peptide/MHC complexes on antigen-presenting cells
(APCs), which will effectively activate these CTL (Alexander-Miller
et al., (1996) Proc Natl Acad Sci USA 93: 4102-7.).
[0007] Recently, HLA class I-binding peptide sequence can be
expected using algorithms (Journal of Immunological Methods,
(1995), Vol. 185, pp. 181-190, J. Immunol., (1994), Vol. 152, pp.
163-175, protein science, (2000), Vol. 9, pp. 1838-1846). However,
it is hard to say that the expected epitope peptide can be cut to
the size and expressed on the target cell surface with HLA molecule
and recognized by CTL. Moreover, the algorithm, for example BIMAS
(http://bimas.dcrt.nih.gov/cgi-bin/molbio/kenparker comboform)
(Parker K C, et al., (1994) J Immunol.; 152(1):163-75.; Kuzushima
K, et al., (2001) Blood.; 98(6):1872-81.)) can suggest the HLA
molecule-binding peptide, but the suggested peptide is not so
rigorous (Bachinsky M M, et. al., Cancer Immun. 2005 Mar. 22;
5:6.). Thus TAA screening still remains a lot of challenges and
difficulties.
[0008] Recent developments in cDNA microarray technologies have
enabled the construction of comprehensive profiles of gene
expression in malignant cells as compared to normal cells (Okabe,
H. et al., (2001) Cancer Res., 61, 2129-37.; Lin Y M. et al.,
(2002) Oncogene, 21; 4120-8.; Hasegawa S. et al., (2002) Cancer Res
62:7012-7.). This approach enables a more thorough understanding of
the complex nature of cancer cells and the mechanisms of
carcinogenesis and facilitates the identification of genes whose
expression is deregulated in tumors (Bienz M. et al., (2000) Cell
103, 311-20.). Among the transcripts identified as up-regulated in
cancers, CDH3 (GenBank Accession No. NM_001793; SEQ ID Nos.1, 2),
EPHA4 (GenBank Accession No. L36645; SEQ ID Nos.3, 4), ECT2
(GenBank Accession No. AY376439; SEQ ID Nos.5, 6), HIG2 (GenBank
Accession No. NM_013332; SEQ ID Nos.7, 8) INHBB (GenBank Accession
No. NM_002193; SEQ ID Nos.9, 435, 10, 436), KIF20A (GenBank
Accession No. NM_005733; SEQ ID Nos.11, 12), KNTC2 (GenBank
Accession No. AF017790; SEQ ID Nos.13, 14), TTK (GenBank Accession
No. NM_003318; SEQ ID Nos.15, 16) and URLC10 (GenBank Accession No.
NM_017527; SEQ ID Nos.17, 18) have been recently discovered. The
entire contents of the references are incorporated by reference
herein. These genes are of particular interest to the present
inventors, being specifically up-regulated in tumor cells of the
various cancer tissues of the cases analyzed (see below). Thus,
immunogenic peptides derived from CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and URLC10 may find utility in selectively
killing tumor cells that express such antigens. The present
invention addresses these and other needs.
[0009] Since cytotoxic drugs, such as M-VAC, often cause severe
adverse reactions, it is clear that thoughtful selection of novel
target molecules on the basis of well-characterized mechanisms of
action should be very helpful in the development of effective
anti-cancer drugs having a minimized risk of side effects. Toward
this goal, expression profile analyses were previously performed on
various cancers and normal human tissue. Such studies led to the
discovery of multiple genes that are specifically over-expressed in
cancer (Lin Y M, et al., Oncogene. 2002 Jun. 13; 21:4120-8.;
Kitahara O, et al., Cancer Res. 2001 May 1; 61:3544-9.; Suzuki C,
et al., Cancer Res. 2003 Nov. 1; 63:7038-41.; Ashida S, Cancer Res.
2004 Sep. 1; 64:5963-72.; Ochi K, et al., Int J Oncol. 2004 March;
24(3):647-55.; Kaneta Y, et al., Int J Oncol. 2003 September;
23:681-91.; Obama K, Hepatology. 2005 June; 41:1339-48.; Kato T, et
al., Cancer Res. 2005 Jul. 1; 65:5638-46.; Kitahara O, et al.,
Neoplasia. 2002 July-August; 4:295-303.; Saito-Hisaminato A et al.,
DNA Res 2002, 9: 35-45.). Examples of such genes identified as
over-expressed in various cancers include, but are not limited to,
CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and URLC10. CDH3
has been previously identified as over-expressed in bladder cancer,
cervical cancer, cholangiocellular carcinoma, colorectal cancer,
endometriosis, gastric cancer, diffuse-type gastric cancer,
non-small cell lung cancer (NSCLC), pancreatic cancer, soft tissue
tumor and testicular tumor. EPHA4 has been identified in bladder
cancer, cervical cancer, cholangiocellular carcinoma,
endometriosis, diffuse-type gastric cancer, ovarian cancer,
pancreatic cancer, prostate cancer and soft tissue tumor. ECT2 has
been identified in bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, chronic myeloid leukemia (CML),
colorectal cancer, esophageal cancer, NSCLC, lymphoma, prostate
cancer, renal carcinoma and small cell lung cancer (SCLC). HIG2 has
been identified in renal carcinoma and SCLC. INHBB has been
identified in cholangiocellular carcinoma, esophageal cancer,
NSCLC, renal carcinoma, SCLC and soft tissue tumor. KIF20A has been
identified in bladder cancer, breast cancer, cholangiocellular
carcinoma, esophageal cancer, NSCLC, pancreatic cancer, prostate
cancer, renal carcinoma and SCLC. KNTC2 has been identified in
bladder cancer, breast cancer, cervical cancer, cholangiocellular
carcinoma, CML, colorectal cancer, esophageal cancer, NSCLC,
lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate
cancer, renal carcinoma, SCLC and soft tissue tumor. TTK has been
identified in bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, CML, colorectal cancer, esophageal
cancer, liver cancer, NSCLC, lymphoma, osteosarcoma, prostate
cancer, SCLC and soft tissue tumor. URLC10 has been identified in
bladder cancer, cervical cancer, cholangiocellular carcinoma,
esophageal cancer, gastric cancer, NSCLC, osteosarcoma, pancreatic
cancer and SCLC.
BRIEF SUMMARY OF THE INVENTION
[0010] The present invention is based in part on the discovery of
the applicable targets of immunotherapy. Because TAAs have often no
immunogenicity, the discovery of appropriate targets is of extreme
importance. As noted above, CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A,
KNTC2, TTK and URLC10 have been identified as up-regulated in
various cancers. More particularly, these genes were identified
using gene expression profiling with a genome-wide cDNA microarray.
As discussed above, expression of CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and URLC10 has been shown to be specifically
up-regulated in various tumor cells, from pancreatic cancer cells
to renal cell carcinomas. As described in Table 1, CDH3 expression
is validly elevated in 26 out of 34 bladder cancer, 17 out of 19
cervical cancer, all of 19 cholangiocellular carcinoma, 30 out of
34 colorectal cancer, 20 out of 21 endometriosis, 13 out of 20
gastric cancer, 7 out of 8 diffuse-type gastric cancer, 36 out of
37 NSCLC, all of 16 pancreatic cancer, all of 21 soft tissue tumor
and all of 10 testicular tumor.
[0011] Table 1 further demonstrates that: [0012] EPHA4 expression
is validly elevated in 14 out of 34 bladder cancer, 8 out of 14
cervical cancer, 10 out of 25 cholangiocellular carcinoma, 5 out of
15 endometriosis, 5 out of 8 diffuse-type gastric cancer, all of 5
ovarian cancer, all 14 pancreatic cancer, 20 out of 51 prostate
cancer and 14 out of 23 soft tissue tumor. [0013] ECT2 expression
is validly elevated in 17 out of 19 bladder cancer, 5 out of 12
breast cancer, all of 14 cervical cancer, all of 13
cholangiocellular carcinoma, all of 5 CML, 7 out of 8 colorectal
cancer, 12 out of 16 esophageal cancer, 6 out of 16 NSCLC, 8 out of
10 lymphoma, 1 out of 1 pancreatic cancer, 10 out of 13 prostate
cancer, 3 out of 6 renal carcinoma and 12 out of 13 SCLC cancer.
[0014] HIG2 expression is validly elevated in 19 out of 20 renal
cancer and 7 out of 9 soft tissue tumor. [0015] INHBB expression is
validly elevated in 10 out of 21 cholangiocellular carcinoma, all
of 12 esophageal cancer, 10 out of 13 NSCLC, 22 out of 24 renal
carcinoma, 8 out of 14 SCLC cancer and 45 out of 49 soft tissue
tumor. [0016] KIF20A expression is validly elevated in all of 31
bladder cancer, 38 out of 61 breast cancer, 10 out of 11
cholangiocellular carcinoma, 7 out of 19 esophageal cancer, 21 out
of 22 NSCLC, all of 6 ovarian cancer, 17 out of 36 prostate cancer,
6 out of 11 renal carcinoma and all of 15 SCLC. [0017] KNTC2
expression is validly elevated in 30 out of 32 bladder cancer, 47
out of 56 breast cancer, all of 10 cervical cancer, 16 out of 22
cholangiocellular carcinoma, 17 out of 37 CML, 3 out of 10
colorectal cancer, 11 out of 46 esophagus cancer, 15 out of 19
NSCLC, 7 out of 8 lymphoma, 20 out of 24 osteosarcoma, 3 out of 5
ovarian cancer, all of 2 pancreatic cancer, 15 out of 37 prostate
cancer, 14 out of 19 renal carcinoma, all of 15 SCLC and 40 out of
59 soft tissue tumor. [0018] TTK expression is validly elevated in
all of 27 bladder cancer, 25 out of 30 breast cancer, 15 out of 16
cervical cancer, all of 10 cholangiocellular carcinoma, 5 out of 7
CML, 6 out of 10 colorectal cancer, 24 out of 44 esophageal cancer,
8 out of 15 liver cancer, all of 12 NSCLC, all of 6 lymphoma, 13
out of 16 osteoblastoma, 12 out of 17 prostate cancer, all of 15
SCLC and 16 out of 33 soft tissue tumor. [0019] URLC10 expression
is validly elevated in all of 29 bladder cancer, 15 out of 16
cervical cancer, all of 7 cholangiocellular carcinoma, 7 out of 19
esophageal cancer, all of 3 gastric cancer, 24 out of 27 NSCLC, 15
out of 19 osteosarcoma, 4 out of 5 pancreatic cancer, 33 out of 43
soft tissue tumor.
[0020] The present invention is based, at least in part, on the
identification of specific epitope peptides of the gene products of
these genes (CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and
URLC10) which possess the ability to induce cytotoxic T lymphocytes
(CTLs) specific to the corresponding molecules. As discussed in
detail below, Peripheral Blood Mononuclear Cells (PBMC) of healthy
donor were stimulated using HLA-A*2402 or HLA-A*0201 binding
candidate peptides derived from CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK or URLC10. CTL clones and/or lines were then
established with specific cytotoxicity against the HLA-A24 or
HLA-A2 (HLA-A02) positive target cells pulsed with each of the
candidate peptides. These results demonstrate that these peptides
are HLA-A24 or HLA-A2 (HLA-A02) restricted epitope peptides that
can induce potent and specific immune responses against cells
expressing CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK or
URLC10.
[0021] Accordingly, the present invention provides methods for
treating or preventing a disease associated with the
over-expression of CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2,
TTK or URLC10, e.g. cancer. Such methods involve the step of
administering to a subject in need thereof a CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10 polypeptides of the
invention. Administration of such peptide(s) results in the
induction of anti-tumor immunity. Thus, the present invention
provides methods for inducing anti-tumor immunity in a subject,
such methods involving the step of administering to the subject the
CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10
polypeptides, as well as pharmaceutical compositions for treating
or preventing a disease associated with the over-expression of
CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10,
e.g. cancer, that include the CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and URLC10 polypeptides. Examples of such
cancers include, but are not limited to, bladder cancer, breast
cancer, cervical cancer, cholangiocellular carcinoma, CML,
colorectal cancer, endometriosis, esophageal cancer, gastric
cancer, diffused type gastric cancer, liver cancer, NSCLC,
lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate
cancer, renal carcinoma, SCLC, soft tissue tumor and testicular
tumor.
[0022] The present invention further provides methods for
preventing post-surgery recurrence of the disease mentioned
above.
[0023] Regarding the specific aims and objectives recited above, it
will be understood by those skilled in the art that one or more
aspects of this invention can meet certain objectives, while one or
more other aspects can meet certain other objectives. Each
objective may not apply equally, in all its respects, to every
aspect of this invention. As such, the objects herein can be viewed
in the alternative with respect to any one aspect of this
invention.
[0024] Additional objects and features of the invention will become
more fully apparent when the following detailed description is read
in conjunction with the accompanying figures and examples. However,
it is to be understood that both the foregoing summary of the
invention and the following detailed description are of preferred
embodiments, and not restrictive of the invention or other
alternate embodiments of the invention. In particular, while the
invention is described herein with reference to a number of
specific embodiments, it will be appreciated that the description
is illustrative of the invention and is not constructed as limiting
of the invention. Various modifications and applications may occur
to those who are skilled in the art, without departing from the
spirit and the scope of the invention, as described by the appended
claims. Likewise, other objects, features, benefits and advantages
of the present invention will be apparent from this summary and
certain embodiments described below, and will be readily apparent
to those skilled in the art. Such objects, features, benefits and
advantages will be apparent from the above in conjunction with the
accompanying examples, data, figures and all reasonable inferences
to be drawn therefrom, alone or with consideration of the
references incorporated herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0025] Various aspects and applications of the present invention
will become apparent to the skilled artisan upon consideration of
the brief description of the figures and the detailed description
of the present invention and its preferred embodiments which
follows:
[0026] FIGS. 1A-1E depict the results of the screening of epitope
peptides, which, in turn, demonstrate that CDH3-A24-10-332 (SEQ ID
NO: 34), CDH3-A24-10-470 (SEQ ID NO: 358), CDH3-A24-9-513 (SEQ ID
NO: 19), CDH3-A24-9-406 (SEQ ID NO: 22), CDH3-A24-10-807 (SEQ ID
NO: 30) and CDH3-A24-10-655 (SEQ ID NO: 344) show potent IFN-gamma
production. FIG. 1A depicts the example of negative peptides which
could not be detected CTL-inducing ability despite possible binding
activity with HLA-A*2402. FIG. 1B depicts the CTL-inducing ability
of CDH3-A24-10-332 (SEQ ID NO: 34). CDH3-A24-10-332 (SEQ ID NO: 34)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line that was established from
the positive well #4 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 1C depicts the CTL-inducing ability of
CDH3-A24-10-470 (SEQ ID NO: 358). CDH3-A24-10-470 (SEQ ID NO: 358)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line that was established from
the positive well #4 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 1D depicts the CTL-inducing ability of CDH3-A24-9-513
(SEQ ID NO: 19). CDH3-A24-9-513 (SEQ ID NO: 19) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay. The well #6 shown in boxed wells in left panel
demonstrated the specific response against the target cells pulsed
with the epitope peptide. Moreover, CTL line that was established
from the positive well #5 shown in boxed wells in middle panel,
demonstrated the specific response against the target cells pulsed
with the epitope peptide. FIG. 1E depicts the CTL-inducing ability
of CDH3-A24-9-406 (SEQ ID NO: 22). CDH3-A24-9-406 (SEQ ID NO: 22)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line that was established from
the positive well #2 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide.
[0027] FIGS. 1F-1G depict the results of the screening of epitope
peptides, which, in turn, demonstrate that CDH3-A24-10-332 (SEQ ID
NO: 34), CDH3-A24-10-470 (SEQ ID NO: 358), CDH3-A24-9-513 (SEQ ID
NO: 19), CDH3-A24-9-406 (SEQ ID NO: 22), CDH3-A24-10-807 (SEQ ID
NO: 30) and CDH3-A24-10-655 (SEQ ID NO: 344) show potent IFN-gamma
production. FIG. 1F depicts the CTL-inducing ability of
CDH3-A24-10-807 (SEQ ID NO: 30). CDH3-A24-10-807 (SEQ ID NO: 30)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and the clone were
established from the positive well #5 shown in boxed wells. The
established CTL clone raised against the peptide demonstrated the
specific CTL activity against COS7 transfected both full length of
CDH3 gene and HLA-A24 molecule (lower right graph). On the other
hand, COS7 transfected full length of CDH3 but not HLA-A24 and COS7
transfected HLA-A24 but not full length of CDH3 were prepared for
the negative control. The CTL clone showed high specific CTL
activity against COS7 that transfected both CDH3 and HLA-A24. FIG.
1G depicts the CTL-inducing ability of CDH3-A24-10-655 (SEQ ID NO:
344). CDH3-A24-10-655 (SEQ ID NO: 344) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line and the clone were established from the
positive well #1 shown in boxed wells. The established CTL clone
raised against the peptide demonstrated the specific CTL activity
against COS7 transfected both full length of CDH3 gene and HLA-A24
molecule (lower right graph). On the other hand, COS7 transfected
full length of CDH3 but not HLA-A24 and COS7 transfected HLA-A24
but not full length of CDH3 were prepared for the negative control.
The CTL clone showed high specific CTL activity against COS7 that
transfected both CDH3 and HLA-A24.
[0028] FIGS. 2A-H depict the results of the screening of epitope
peptides, which, in turn, demonstrate that Epha4-A24-9-453 (SEQ ID
NO: 41), Epha4-A24-9-5 (SEQ ID NO: 44), Epha4-A24-9-420 (SEQ ID NO:
48), Epha4-A24-9-869 (SEQ ID NO: 46), Epha4-A24-10-24 (SEQ ID NO:
78) Epha4-A02-9-501 (SEQ ID NO: 376) and Epha4-A02-9-165 (SEQ ID
NO: 379) show potent IFN-gamma production. FIG. 2A depicts the
example of negative peptides which could not be detected
CTL-inducing ability despite possible binding activity with HLA.
FIG. 2B depicts the CTL-inducing ability of Epha4-A24-9-453 (SEQ ID
NO: 41). Epha4-A24-9-453 (SEQ ID NO: 41) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line that was established from the positive
well #3 shown in boxed wells, demonstrated the specific response
against the target cells pulsed with the epitope peptide. FIG. 2C
depicts the CTL-inducing ability of Epha4-A24-9-5 (SEQ ID NO: 44).
Epha4-A24-9-5 (SEQ ID NO: 44) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay,
and CTL line, that was established from the positive well #2 shown
in boxed wells, demonstrated the specific response against the
target cells pulsed with the epitope peptide. FIG. 2D depicts the
CTL-inducing ability of Epha4-A24-9-420 (SEQ ID NO: 48).
Epha4-A24-9-420 (SEQ ID NO: 48) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay.
The well #6 shown in boxed wells in upper panel demonstrated the
specific response against the target cells pulsed with the epitope
peptide. Moreover CTL line that was established from the positive
well #6 shown in boxed wells in middle panel, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 2E depicts the CTL-inducing ability of
Epha4-A24-9-869 (SEQ ID NO: 46). Epha4-A24-9-869 (SEQ ID NO: 46)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line that was established from
the positive well #5 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 2F depicts the CTL-inducing ability of
Epha4-A24-10-24 (SEQ ID NO: 78). Epha4-A24-10-24 (SEQ ID NO: 78)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line that was established from
the positive well #4 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 2G depicts the CTL-inducing ability of
Epha4-A02-9-501 (SEQ ID NO: 376). Epha4-A02-9-501 (SEQ ID NO: 376)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and clone was established
from the positive well #8 shown in boxed wells. Cytotoxic activity
of the established CTL line against the target cells pulsed with
the peptide was measured by Cr-release assay (CRA) (lower graph),
and the CTL line had very potent specific cytotoxic activity
against the target cells pulsed with the peptides. FIG. 2H depicts
the CTL-inducing ability of Epha4-A02-9-165 (SEQ ID NO: 379).
Epha4-A02-9-165 (SEQ ID NO: 379) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay,
and CTL line was established from the positive well #3 shown in
boxed wells. Cytotoxic activity of the established CTL line against
target cells pulsed with peptide was measured by Cr-release assay
(CRA) (right graph), and the CTL line had very potent specific
cytotoxic activity against the target cells pulsed with the
peptides.
[0029] FIGS. 3A-3D depict the results of the screening of epitope
peptides, which, in turn, demonstrate that ECT2-A24-9-515 (SEQ ID
NO: 80), ECT2-A24-10-40 (SEQ ID NO: 100) and ECT2-A24-10-101 (SEQ
ID NO: 101) show potent IFN-gamma production. FIG. 3A depicts the
example of negative peptides which could not be detected
CTL-inducing ability despite possible binding activity with HLA.
FIG. 3B depicts the CTL-inducing ability of ECT2-A24-9-515 (SEQ ID
NO: 80). ECT2-A24-9-515 (SEQ ID NO: 80) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay. The well #5 and #7 shown in boxed wells in left
panel demonstrated the specific response against the target cells
pulsed with the epitope peptide. Moreover, CTL line that was
established from the positive well #7 shown in boxed wells in
second panel, demonstrated the specific response against the target
cells pulsed with the epitope peptide. Cytotoxic activity of the
CTL line against cancer cell line, TE6 endogenously expressing ECT2
and HLA-A24 was measured by Cr-release assay (CRA), and the CTL
clone had very potent cytotoxic activity against TE6. On the other
hand, the effector cells did not demonstrate the cytotoxic activity
of the CTL line against cancer cell line, TE5 expressing only ECT2
was not detected. FIG. 3C depicts the CTL-inducing ability of
ECT2-A24-10-40 (SEQ ID NO: 100). ECT2-A24-10-40 (SEQ ID NO: 100)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and the clone were
established from the positive well #2 shown in boxed wells. The
established CTL clone raised against the peptide demonstrated
specific CTL activity against COS7 transfected both full length of
ECT2 gene and HLA-A24 molecule. On the other hand, COS7 transfected
full length of ECT2 but not HLA-A24, COS7 transfected HLA-A24 and
URLC10 gene as a substitute for full length of ECT2 and COS7
transfected HLA-A24 and pulsed with ECT2-10-101 were prepared for
the negative control. The CTL clone showed high specific CTL
activity against COS7 that transfected both ECT2 and HLA-A24. "d"
depicts the CTL-inducing ability of ECT2-A24-10-101 (SEQ ID NO:
101). ECT2-A24-10-101 (SEQ ID NO: 101) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line were established from the positive well
#1 shown in boxed wells. The established CTL line raised against
the peptide demonstrated specific CTL activity against COS7
transfected both full length of ECT2 gene and HLA-A24 molecule.
COS7 transfected full length of ECT2 but not HLA-A24, COS7
transfected HLA-A24 and URLC10 gene as substitute for full length
of ECT2 and COS7 transfected HLA-A24 and pulsed with ECT2-10-40
were prepared for the negative control. The CTL clone showed high
specific CTL activity against COS7 that transfected both ECT2 and
HLA-A24.
[0030] FIGS. 4A-4E depict the results of the screening of epitope
peptides, which, in turn, demonstrate that HIG2-A24-9-19 (SEQ ID
NO: 110), HIG2-A24-9-22 (SEQ ID NO: 111), HIG2-A24-9-8 (SEQ ID NO:
387), HIG2-A24-10-7 (SEQ ID NO: 112), HIG2-A24-10-18 (SEQ ID NO:
394), HIG2-A02-9-15 (SEQ ID NO: 116), HIG2-A02-9-4 (SEQ ID NO: 117)
and HIG2-A02-10-8 (SEQ ID NO: 121) show potent IFN-gamma
production. FIG. 4A depicts the example of negative peptides which
could not be detected CTL-inducing ability despite possible binding
activity with HLA. FIG. 4B depicts the CTL-inducing ability of
HIG2-A24-9-19 (SEQ ID NO: 110). HIG2-A24-9-19 (SEQ ID NO: 110)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line, that was established from
the positive well #6 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 4C depicts the CTL-inducing ability of HIG2-A24-9-22
(SEQ ID NO: 111). HIG2-A24-9-22 (SEQ ID NO: 111) demonstrated
potent IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line and clone, that was established from
the positive well #7 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 4D depicts the CTL-inducing ability of HIG2-A24-9-8
(SEQ ID NO: 387). HIG2-A24-9-8 (SEQ ID NO: 387) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line and clone, that were established from
the positive well #5 shown in boxed wells, demonstrated the
specific response against the target cells pulsed with the epitope
peptide. FIG. 4E depicts the CTL-inducing ability of HIG2-A02-9-8
(SEQ ID NO: 114). HIG2-A02-9-8 (SEQ ID NO: 114) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line was established from the positive well
#10 shown in boxed wells. The established CTL line raised against
the peptide demonstrate specific CTL activity against 293T
transfected both full length of HIG2 gene and HLA-A02 molecule.
293T transfected full length of HIG2 but not HLA-A02, 293 Ts
transfected HLA-A02 and FoxP3 gene as substitute for full length of
HIG2 and 293 Ts transfected HLA-A02 and pulsed with HIG2-9-15 were
prepared for the negative control. The CTL line showed high
specific CTL activity against 293T that transfected both HIG2 and
HLA-A02.
[0031] FIGS. 4F-4H depict the results of the screening of epitope
peptides, which, in turn, demonstrate that HIG2-A24-9-19 (SEQ ID
NO: 110), HIG2-A24-9-22 (SEQ ID NO: 111), HIG2-A24-9-8 (SEQ ID NO:
387), HIG2-A24-10-7 (SEQ ID NO: 112), HIG2-A24-10-18 (SEQ ID NO:
394), HIG2-A02-9-15 (SEQ ID NO: 116), HIG2-A02-9-4 (SEQ ID NO: 117)
and HIG2-A02-10-8 (SEQ ID NO: 121) show potent IFN-gamma
production. FIG. 4F depicts the CTL-inducing ability of
HIG2-A24-10-7 (SEQ ID NO: 112). HIG2-A24-10-7 (SEQ ID NO: 112)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL lines or clone, that were
established from the positive well #1 and #7 shown in boxed wells,
demonstrated the specific response against the target cells pulsed
with the epitope peptide. FIG. 4G depicts the CTL-inducing ability
of HIG2-A24-10-18 (SEQ ID NO: 394). HIG2-A24-10-18 (SEQ ID NO: 394)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and clone, that were
established from the positive well #7 shown in boxed wells,
demonstrated the specific response against the target cells pulsed
with the epitope peptide. FIG. 4H depicts the CTL-inducing ability
of HIG2-A02-9-15 (SEQ ID NO: 116). HIG2-A02-9-15 (SEQ ID NO: 116)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line was established from the
positive well #10 shown in boxed wells. The established CTL line
raised against the peptide demonstrated specific CTL activity
against COS7 transfected both full length of HIG2 gene and HLA-A02
molecule. COS7 transfected full length of HIG2 but not HLA-A02 and
COS7s transfected HLA-A02 and pulsed with HIG2-9-8 peptide were
prepared for the negative control. The CTL line showed high
specific CTL activity against COS7 that transfected both HIG2 and
HLA-A02.
[0032] FIGS. 4I-4J depict the results of the screening of epitope
peptides, which, in turn, demonstrate that HIG2-A24-9-19 (SEQ ID
NO: 110), HIG2-A24-9-22 (SEQ ID NO: 111), HIG2-A24-9-8 (SEQ ID NO:
387), HIG2-A24-10-7 (SEQ ID NO: 112), HIG2-A24-10-18 (SEQ ID NO:
394), HIG2-A02-9-15 (SEQ ID NO: 116), HIG2-A02-9-4 (SEQ ID NO: 117)
and HIG2-A02-10-8 (SEQ ID NO: 121) show potent IFN-gamma
production. FIG. 4I depicts the CTL-inducing ability of
HIG2-A02-9-4 (SEQ ID NO: 117). HIG2-A02-9-4 (SEQ ID NO: 117)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and clone were established
from the positive well #10 shown in boxed wells. The established
CTL line raised against the peptide demonstrated specific CTL
activity against COS7 transfected both full length of HIG2 gene and
HLA-A02 molecule (middle graph). Also, COST transfected full length
of HIG2 but not HLA-A02, COS7s transfected HLA-A02 and TTK gene as
substitute for full length of HIG2 and COS7s transfected HLA-A02
and pulsed with HIG2-9-8 were prepared for the negative control.
Cytotoxic activity of the CTL clone against 293T, transfected both
full length of HIG2 gene and HLA-A02 molecule, and cancer cell
line, Caki-1 endogenously expressing HIG2 and HLA-A02 was measured
by Cr-release assay (CRA) (lower graphs), and the CTL clone had
very potent cytotoxic activity against the transfectant with both
of HIG2 gene and HLA-A02, and Caki-1. On the other hand, the
effector cells did not demonstrate the cytotoxic activity of the
CTL line against 293T, transfected only HIG2 or only HLA-A02, and
cancer cell line, A498 expressing only HIG2 was not detected. FIG.
4J depicts the CTL-inducing ability of HIG2-A02-10-8 (SEQ ID NO:
121). HIG2-A02-10-8 (SEQ ID NO: 121) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay,
and CTL line, that was established from the positive well #9 shown
in boxed wells, demonstrated the specific response against the
target cells pulsed with the epitope peptide.
[0033] FIGS. 5A-5C depict the results of the screening of epitope
peptides, which, in turn, demonstrate that INHBB-A24-9-180 (SEQ ID
NO: 395), INHBB-A24-10-180 (SEQ ID NO: 133), INHBB-A24-10-305 (SEQ
ID NO: 135), INHBB-A24-10-7 (SEQ ID NO: 137) and INHBB-A24-10-212
(SEQ ID NO: 426) show potent IFN-gamma production. FIG. 5A depicts
the example of negative peptides which could not be detected
CTL-inducing ability despite possible binding activity with HLA.
FIG. 5B depicts the CTL-inducing ability of INHBB-A24-9-180 (SEQ ID
NO: 395). INHBB-A24-9-180 (SEQ ID NO: 395) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line and clone was established from the
positive well #7 shown in boxed wells. Cytotoxic activity of the
established CTL clone against tumor cells, Miapaca2 expressing both
of INHBB and HLA-A02 was measured by Cr-release assay (CRA), and
the effector cells showed high specific cytotoxic activity against
Miapaca2. On the other hand, it did not show significant specific
cytotoxic activity against Caki-1 expressing INHBB but not HLA-A02.
FIG. 5C depicts the CTL-inducing ability of INHBB-A24-10-180 (SEQ
ID NO: 133). INHBB-A24-10-180 (SEQ ID NO: 133) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line was established from the positive well
#3 shown in boxed wells. The established CTL line raised against
the peptide demonstrated high specific CTL activity against 293T
transfected both of full length of INHBB gene and HLA-A24 molecule.
Also, 293T transfected full length of INHBB but not HLA-A24 and 293
Ts transfected HLA-A24 and pulsed with INHBB-10-305 peptide were
prepared for the negative control.
[0034] FIGS. 5D-5F depict the results of the screening of epitope
peptides, which, in turn, demonstrate that INHBB-A24-9-180 (SEQ ID
NO: 395), INHBB-A24-10-180 (SEQ ID NO: 133), INHBB-A24-10-305 (SEQ
ID NO: 135), INHBB-A24-10-7 (SEQ ID NO: 137) and INHBB-A24-10-212
(SEQ ID NO: 426) show potent IFN-gamma production. FIG. 5D depicts
the CTL-inducing ability of INHBB-A24-10-305 (SEQ ID NO: 135).
INHBB-A24-10-305 (SEQ ID NO: 135) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay,
and CTL line and clone were established from the positive well #2
shown in boxed wells. The established CTL clone raised against the
peptide demonstrated high specific CTL activity against 293T
transfected both full length of INHBB gene and HLA-A24 molecule.
Also, 293T transfected full length of INHBB but HLA-A24 and 293 Ts
transfected HLA-A24 and pulsed with INHBB-10-180 peptide were
prepared for the negative control. FIG. 5E depicts the CTL-inducing
ability of INHBB-A24-10-7 (SEQ ID NO: 137)). INHBB-A24-10-7 (SEQ ID
NO: 137) demonstrated potent IFN-gamma production as compared to
the control by IFN-gamma ELISPOT assay, and CTL lines were
established from the positive well #8 shown in boxed wells in upper
panel and #2 shown in boxed wells in lower panel. The CTL line from
#8 well demonstrated specific CTL activity against 293T transfected
both full length of INHBB gene and HLA-A24 molecule. Also, 293T
transfected full length of INHBB but not HLA-A24 and 293 Ts
transfected HLA-A24 and pulsed with INHBB-10-40 peptide were
prepared for the negative control. FIG. 5F depicts the CTL-inducing
ability of INHBB-A24-10-212 (SEQ ID NO: 426). INHBB-A24-10-212 (SEQ
ID NO: 426) demonstrated potent IFN-gamma production as compared to
the control by IFN-gamma ELISPOT assay, and CTL line, that was
established from the positive well #1 shown in boxed wells,
demonstrated the specific response against the target cells pulsed
with the epitope peptide.
[0035] FIGS. 6A-6E depict the results of the screening of epitope
peptides, which, in turn, demonstrate that KIF20A-A24-10-304 (SEQ
ID NO: 186), KIF20A-A24-9-383 (SEQ ID NO: 178), KIF20A-A24-10-66
(SEQ ID NO: 194) and KIF20A-A24-9-305 (SEQ ID NO: 174) show potent
IFN-gamma production. FIG. 6 A depicts the example of negative
peptides which could not be detected CTL-inducing ability despite
possible binding activity with HLA. FIG. 6 B depicts the
CTL-inducing ability of KIF20A-A24-10-304 (SEQ ID NO: 186).
KIF20A-A24-10-304 (SEQ ID NO: 186) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay.
The well #5 shown in boxed wells in lower right panel demonstrated
the specific response against the target cells pulsed with the
epitope peptide. Moreover, CTL line and clone, that were
established from the positive well #5 shown in boxed wells in upper
left panel, also demonstrated the specific response against the
target cells pulsed with the epitope peptide. The established CTL
clone raised against the peptide demonstrated specific CTL activity
against 24-LCL transfected full length of KIF20A gene. Also,
A24-LCL transfected mock vector was prepared for the negative
control. Cytotoxic activity of the CTL clone against tumor cells,
Miapaca2 expressing both of KIF20A and HLA-A24 was measured by
Cr-release assay (CRA), and the CTL clone had very potent specific
cytotoxic activity against Miapaca2 (lower right graph). On the
other hand, it did not show significant specific cytotoxic activity
against PK59 expressing KIF20A but not HLA-A24. FIG. 6C depicts the
CTL-inducing ability of KIF20A-A24-9-383 (SEQ ID NO: 178).
KIF20A-A24-9-383 (SEQ ID NO: 178) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay.
The well #3 and 4 shown in boxed wells in right panel demonstrated
the specific response against the target cells pulsed with the
epitope peptide. Moreover, CTL line, that was established from the
positive well #3 shown in boxed wells in left panel, also
demonstrated the specific response against the target cells pulsed
with the epitope peptide. The established CTL line demonstrated
high specific CTL activity against COS7 transfected both full
length of KIF20A gene and HLA-A24 molecule. Also, COS7 transfected
full length of KIF20A but not HLA-A24 and COS7s transfected HLA-A24
and pulsed with KIF20A-9-621 peptide were prepared for the negative
control.
[0036] FIGS. 6D-6E depict the results of the screening of epitope
peptides, which, in turn, demonstrate that KIF20A-A24-10-304 (SEQ
ID NO: 186), KIF20A-A24-9-383 (SEQ ID NO: 178), KIF20A-A24-10-66
(SEQ ID NO: 194) and KIF20A-A24-9-305 (SEQ ID NO: 174) show potent
IFN-gamma production. FIG. 6D depicts the CTL-inducing ability of
KIF20A-A24-10-66 (SEQ ID NO: 194). KIF20A-A24-10-66 (SEQ ID NO:
194) demonstrated potent IFN-gamma production as compared to the
control by IFN-gamma ELISPOT assay, and CTL lines, that were
established from the positive well #6 shown in boxed wells in upper
left panel and #3 shown in boxed wells in lower middle panel
demonstrated the specific response against the target cells pulsed
with the epitope peptide. Moreover, CTL clone selected from CTL
line from #6 well by limiting dilution demonstrated specific CTL
activity against the target cells. The established CTL clone showed
specific CTL activity against COS7 transfected both full length of
KIF20A gene and HLA-A24 molecule. Also, COS7 transfected full
length of KIF20A but not HLA-A24, COS7s transfected HLA-A24 and
URLC10 gene as substitute for full length of KIF20A and COS7
transfected HLA-A24 and pulsed with KIF20A-10-308 peptide were
prepared for the negative control. FIG. 6E depicts the CTL-inducing
ability of KIF20A-A24-9-305 (SEQ ID NO: 174). KIF20A-A24-9-305 (SEQ
ID NO: 174) demonstrated potent IFN-gamma production as compared to
the control by IFN-gamma ELISPOT assay, and CTL lines, that were
established from the positive well #2 shown in boxed wells in upper
left panel and #6 shown in boxed wells in lower middle panel,
demonstrated the specific response against the target cells pulsed
with the epitope peptide. Moreover, CTL clone selected from CTL
line from #2 well by limiting dilution demonstrated specific CTL
activity against the target cells. Cytotoxic activity of the CTL
clone against tumor cells, PK45P expressing both of KIF20A and
HLA-A24 was measured by Cr-release assay (CRA), and the CTL clone
had very potent cytotoxic activity against PK45P. On the other
hand, it did not show significant specific cytotoxic activity
against PK59 expressing KIF20A but not HLA-A24.
[0037] FIGS. 7A-7E depict the results of the screening of epitope
peptides, which, in turn, demonstrate that KNTC2-A24-9-309 (SEQ ID
NO: 196), KNTC2-A24-9-124 (SEQ ID NO: 202), KNTC2-A24-9-154 (SEQ ID
NO: 210) KNTC2-A24-9-150 (SEQ ID NO: 213), KNTC2-A24-10-452 (SEQ ID
NO: 214), KNTC2-A24-10-227 (SEQ ID NO: 217) and KNTC2-A24-10-273
(SEQ ID NO: 223) show potent IFN-gamma production. FIG. 7A depicts
the example of negative peptides which could not be detected
CTL-inducing ability despite possible binding activity with HLA.
FIG. 7B depicts the CTL-inducing ability of KNTC2-A24-9-309 (SEQ ID
NO: 196). KNTC2-A24-9-309 (SEQ ID NO: 196) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line, that was established from the positive
well #8 shown in boxed wells, demonstrated the specific response
against the target cells pulsed with the epitope peptide. FIG. 7C
depicts the CTL-inducing ability of KNTC2-A24-9-124 (SEQ ID NO:
202). KNTC2-A24-9-124 (SEQ ID NO: 202) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line, that was established from the positive
well #5 shown in boxed wells, demonstrated the specific response
against the target cells pulsed with the epitope peptide. FIG. 7D
depicts the CTL-inducing ability of KNTC2-A24-9-154 (SEQ ID NO:
210). KNTC2-A24-9-154 (SEQ ID NO: 210) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line and clone, that were established from
the positive well #5 shown in boxed wells demonstrated the specific
response against the target cells pulsed with the epitope peptide.
FIG. 7E depicts the CTL-inducing ability of KNTC2-A24-9-150 (SEQ ID
NO: 213). KNTC2-A24-9-150 (SEQ ID NO: 213) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line, that was established from the positive
well #7 shown in boxed wells, demonstrated the specific response
against the target cells pulsed with the epitope peptide.
[0038] FIGS. 7F-7H depict the results of the screening of epitope
peptides, which, in turn, demonstrate that KNTC2-A24-9-309 (SEQ ID
NO: 196), KNTC2-A24-9-124 (SEQ ID NO: 202), KNTC2-A24-9-154 (SEQ ID
NO: 210) KNTC2-A24-9-150 (SEQ ID NO: 213), KNTC2-A24-10-452 (SEQ ID
NO: 214), KNTC2-A24-10-227 (SEQ ID NO: 217) and KNTC2-A24-10-273
(SEQ ID NO: 223) show potent IFN-gamma production. FIG. 7F depicts
the CTL-inducing ability of KNTC2-A24-10-452 (SEQ ID NO: 214).
KNTC2-A24-10-452 (SEQ ID NO: 214) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay,
and CTL lines and clone, that were established from the positive
well #4 shown in boxed wells in upper left panel and #5 shown in
boxed wells in middle panel, demonstrated the specific response
against the target cells pulsed with the epitope peptide. Moreover,
CTL clone selected from CTL line from #5 well by limiting dilution
demonstrated specific CTL activity against the target cells. The
established CTL line from #4 well showed specific CTL activity
against HEK293 transfected both full length of KNTC2 gene and
HLA-A24 molecule. Also, HEK293 transfected full length of KNTC2 but
not HLA-A24, HEK293 transfected HLA-A24 but full length of KNTC2
and HEK293 transfected HLA-A24 pulsed with KNTC-9-309 peptide were
prepared for the negative control. FIG. 7G depicts the CTL-inducing
ability of KNTC2-A24-10-227 (SEQ ID NO: 217). KNTC2-A24-10-227 (SEQ
ID NO: 217) demonstrated potent IFN-gamma production as compared to
the control by IFN-gamma ELISPOT assay, and CTL line, that was
established from the positive well #1 shown in boxed well s,
demonstrated the specific response against the target cells pulsed
with the epitope peptide. FIG. 7H depicts the CTL-inducing ability
of KNTC2-A24-10-273 (SEQ ID NO: 223). KNTC2-A24-10-273 (SEQ ID NO:
223) demonstrated potent IFN-gamma production as compared to the
control by IFN-gamma ELISPOT assay, and CTL line, that was
established from the positive well #8 shown in boxed wells,
demonstrated the specific response against the target cells pulsed
with the epitope peptide.
[0039] FIGS. 8A-8C depict the results of the screening of epitope
peptides, which, in turn, demonstrate that TTK-A02-9-462 (SEQ ID
NO: 227), TTK-A02-9-719 (SEQ ID NO: 233), TTK-A02-9-547 (SEQ ID NO:
228) and TTK-A02-10-462 (SEQ ID NO: 254), show potent IFN-gamma
production. FIG. 8A depicts the example of negative peptides which
could not be detected CTL-inducing ability despite possible binding
activity with HLA. FIG. 8B depicts the CTL-inducing ability of
TTK-A02-9-462 (SEQ ID NO: 227). TTK-A02-9-462 (SEQ ID NO: 227)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and two clones, that were
established from the positive well #4 shown in boxed wells,
demonstrated the specific response against the target cells pulsed
with the epitope peptide. The established CTL clone showed high
specific CTL activity against COS7 transfected both full length of
TTK gene and HLA-A02 molecule. Also, COS7 transfected full length
of TTK but not HLA-A02, COS7s transfected HLA-A02 but not full
length of TTK and COS7s transfected HLA-A02 pulsed with TTK-9-547
peptide were prepared for the negative control. FIG. 8C depicts the
CTL-inducing ability of TTK-A02-9-719 (SEQ ID NO: 233).
TTK-A02-9-719 (SEQ ID NO: 233) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay,
and CTL line and clones were established from the positive well #1
shown in boxed wells. The established CTL line showed high specific
CTL activity against COS7 transfected both full length of TTK gene
and HLA-A02 molecule. Also, COS7 transfected full length of TTK but
not HLA-A02 and COS7s transfected HLA-A02 and HIG2 gene as
substitute for full length of TTK were prepared for the negative
control.
[0040] FIG. 8D depicts the results of the screening of epitope
peptides, which, in turn, demonstrate that TTK-A02-9-462 (SEQ ID
NO: 227), TTK-A02-9-719 (SEQ ID NO: 233), TTK-A02-9-547 (SEQ ID NO:
228) and TTK-A02-10-462 (SEQ ID NO: 254), show potent IFN-gamma
production. FIG. 8D depicts the CTL-inducing ability of
TTK-A02-9-547 (SEQ ID NO: 228). TTK-A02-9-547 (SEQ ID NO: 228)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and clones were
established from the positive well #2 shown in boxed wells. The
established CTL line showed specific CTL activity against COS7
transfected both full length of TTK gene and HLA-A02 molecule.
Also, COS7 transfected full length of TTK but not HLA-A02, COS7s
transfected HLA-A02 but not full length of TTK and COS7s
transfected HLA-A02 and pulsed with TTK-10-462 were prepared for
the negative control.
[0041] FIG. 8E depicts the results of the screening of epitope
peptides, which, in turn, demonstrate that TTK-A02-9-462 (SEQ ID
NO: 227), TTK-A02-9-719 (SEQ ID NO: 233), TTK-A02-9-547 (SEQ ID NO:
228) and TTK-A02-10-462 (SEQ ID NO: 254), show potent IFN-gamma
production. FIG. 8E depicts the CTL-inducing ability of
TTK-A02-10-462 (SEQ ID NO: 254). TTK-A02-10-462 (SEQ ID NO: 254)
demonstrated potent IFN-gamma production as compared to the control
by IFN-gamma ELISPOT assay, and CTL line and three clones were
established from the positive well #8 shown in boxed wells. The
established CTL clone showed specific CTL activity against COS7
transfected both full length of TTK gene and HLA-A02 molecule.
Also, COS7 transfected full length of TTK but not HLA-A02, COS7s
transfected HLA-A02 but not full length of TTK and COS7s
transfected HLA-A02 and pulsed with TTK-9-547 peptide were prepared
for the negative control.
[0042] FIGS. 9A-9D depict the results of the screening of epitope
peptides, which, in turn, demonstrate that URLC10-A02-9-206 (SEQ ID
NO: 271), URLC10-A02-9-212 (SEQ ID NO: 272) and URLC10-A02-10-211
(SEQ ID NO: 288) show potent IFN-gamma production. FIG. 9A depicts
the example of negative peptides which could not be detected
CTL-inducing ability despite possible binding activity with HLA.
FIG. 9B depicts the CTL-inducing ability of URLC10-A02-9-206 (SEQ
ID NO: 271). URLC10-A02-9-206 (SEQ ID NO: 271) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line, that was established from the positive
well #7 shown in boxed wells, demonstrated the specific response
against the target cells pulsed with the epitope peptide. FIG. 9C
depicts the CTL-inducing ability of URLC10-A02-9-212 (SEQ ID NO:
272). URLC10-A02-9-212 (SEQ ID NO: 272) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line, that was established from the positive
well #3 shown in boxed wells, demonstrated the specific response
against the target cells pulsed with the epitope peptide. FIG. 9D
depicts the CTL-inducing ability of URLC10-A02-10-211 (SEQ ID NO:
288). URLC10-A02-10-211 (SEQ ID NO: 288) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay, and CTL line and clones, were established from the
positive well #5 shown in boxed wells.
[0043] FIG. 9D (continuation) depicts the results of the screening
of epitope peptides, which, in turn, demonstrate that
URLC10-A02-9-206 (SEQ ID NO: 271), URLC10-A02-9-212 (SEQ ID NO:
272) and URLC10-A02-10-211 (SEQ ID NO: 288) show potent IFN-gamma
production. FIG. 9D (continuation) The established CTL clone showed
high specific CTL activity against COS7, Hek293 and 293T which were
transfected both full length of URLC10 gene and HLA-A02 molecule.
Also, COS7, Hek293 or 293T which were transfected full length of
URLC10 but not HLA-A02 and COS7s, Hek293s or 293 Ts, which were
transfected HLA-A02 and pulsed with URLC10-10-64, were prepared for
the negative control. In this drawings, "+" means the peptide
pulsed target, "-" means the no peptide pulsed target, "R" means
Responder, "S" means Stimulator, "E" means Effector, and "T" means
Target.
[0044] FIGS. 10A-10N include a series of photographs, (a) (n),
depicting the results of IFN-gamma ELISPOT assay on CTLs that were
induced with peptides derived from INHBB. The CTLs in well #4
stimulated with INHBB-A02-9-213 (SEQ ID NO: 143) FIG. 10A, well #5
and #7 stimulated with INHBB-A02-9-174 (SEQ ID NO: 147) FIG. 10B,
well #8 stimulated with INHBB-A02-9-257 (SEQ ID NO: 148) FIG. 10C,
well #1 and #8 stimulated with INHBB-A02-9-313 (SEQ ID NO: 149)
FIG. 10D, well #1, #4 and #8 stimulated with INHBB-A02-9-139 (SEQ
ID NO: 150) FIG. 10E, well #4 stimulated with INHBB-A02-9-8 (SEQ ID
NO: 152) FIG. 10F, well #6 stimulated with INHBB-A02-9-250 (SEQ ID
NO: 153) FIG. 10G, well #5 stimulated with INHBB-A02-10-179 (SEQ ID
NO: 154) FIG. 10H, well #3 stimulated with INHBB-A02-10-237 (SEQ ID
NO: 156) FIG. 10I, well #5 stimulated with INHBB-A02-10-313 (SEQ ID
NO: 160) FIG. 10J, well #3 and #7 stimulated with INHBB-A02-10-173
(SEQ ID NO: 161) FIG. 10K, well #4 stimulated with INHBB-A02-10-256
(SEQ ID NO: 162) FIG. 10L, well #7 stimulated with INHBB-A02-10-162
(SEQ ID NO: 163) FIG. 10M and well #7 stimulated with
INHBB-A02-10-85 (SEQ ID NO: 166) FIG. 10N showed potent IFN-gamma
production as compared with the control respectively. In the
figures, "+" indicates that the target cells in the well were
pulsed with the appropriate peptide, and "-" indicates that the
target cells had not been pulsed with any peptides.
[0045] FIG. 11 depicts a line graph showing the results of
establishment of CTL lines stimulated with INHBB-A02-9-174 (SEQ ID
NO: 147) with IFN-gamma ELISA assay. The depicted results
demonstrate that CTL line established by stimulation with the
peptide showed potent IFN-gamma production as compared with the
control. In the figures, "+" indicates that the target cells were
pulsed with the appropriate peptide and "-" indicates that the
target cells had not been pulsed with any peptides.
DETAILED DESCRIPTION OF THE INVENTION
[0046] The words "a", "an", and "the" as used herein mean "at least
one" unless otherwise specifically indicated. Unless otherwise
defined, all technical and scientific terms used herein have the
same meaning as commonly understood by one of ordinary skill in the
art to which this invention belongs.
[0047] The present invention is based in part on the discovery of
applicable targets of immunotherapy. Identification of new TAAs,
particularly those that induce potent and specific anti-tumor
immune responses, warrants further development of the clinical
application of the peptide vaccination strategy in various types of
cancer (Boon T et al., (1996) J Exp Med 183: 725-9.; van der
Bruggen P et al., (1991) Science 254: 1643-7.; Brichard V et al.,
(1993) J Exp Med 178: 489-95.; Kawakami Y et al., (1994) J Exp Med
180: 347-52.; Shichijo S et al., (1998) J Exp Med 187:277-88.; Chen
Y T et al., (1997) Proc. Natl. Acad. Sci. USA, 94: 1914-8.; Harris
C C, (1996) J Natl Cancer Inst 88:1442-55.; Butterfield L H et al.,
(1999) Cancer Res 59:3134-42.; Vissers J L et al., (1999) Cancer
Res 59: 5554-9.; van der Burg S H et al., (1996) J. Immunol
156:3308-14.; Tanaka F et al., (1997) Cancer Res 57:4465-8.; Fujie
T et al., (1999) Int J Cancer 80:169-72.; Kikuchi M et al., (1999)
Int J Cancer 81: 459-66.; Oiso M et al., (1999) Int J Cancer
81:387-94.). Because TAAs have often no immunogenicity, discovery
of fitting targets is extremely important issue.
[0048] As noted above, [0049] CDH3 (GenBank Accession No.
NM_001793; SEQ ID Nos.1, 2), [0050] EPHA4 (GenBank Accession No.
L36645; SEQ ID Nos.3, 4), [0051] ECT2 (GenBank Accession No.
AY376439; SEQ ID Nos.5, 6), [0052] HIG2 (GenBank Accession No.
NM_013332; SEQ ID Nos.7, 8) [0053] INHBB (GenBank Accession No.
NM_002193; SEQ ID Nos.9, 435, 10, 436), [0054] KIF20A (GenBank
Accession No. NM_005733; SEQ ID Nos.11, 12), [0055] KNTC2 (GenBank
Accession No. AF017790; SEQ ID Nos.13, 14), [0056] TTK (GenBank
Accession No. NM_003318; SEQ ID Nos.15, 16) and [0057] URLC10
(GenBank Accession No. NM_017527; SEQ ID Nos.17, 18) were
previously identified as over-expressed in various cancers using
cDNA microarray technologies.
[0058] In the present invention, peptides derived from CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK or URLC10 are shown to be TAA
epitopes restricted by HLA-A24 and HLA-A2 (HLA-A02), an HLA allele
commonly found in the Japanese and Caucasian populations.
Specifically, using their binding affinities to HLA-A24 or HLA-A2
(HLA-A02), candidates of HLA-A24 or HLA-A2 (HLA-A02) binding
peptides derived from CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A,
KNTC2, TTK or URLC10 were identified. After the in vitro
stimulation of T-cells by dendritic cells (DCs) loaded with these
peptides, CTLs were successfully established using the following
peptides. [0059] CDH3-A24-9-513 (SEQ ID NO: 19), [0060]
CDH3-A24-9-406 (SEQ ID NO: 22), [0061] CDH3-A24-10-807 (SEQ ID NO:
30), [0062] CDH3-A24-10-332 (SEQ ID NO: 34), [0063] CDH3-A24-10-655
(SEQ ID NO: 344), [0064] CDH3-A24-10-470 (SEQ ID NO: 358), [0065]
EphA4-A24-9-453 (SEQ ID NO: 41), [0066] EphA4-A24-9-5 (SEQ ID NO:
44), [0067] EphA4-A24-9-869 (SEQ ID NO: 46), [0068] EphA4-A24-9-420
(SEQ ID NO: 48), [0069] EphA4-A24-10-24 (SEQ ID NO: 78), [0070]
EphA4-A02-9-501 (SEQ ID NO: 376), [0071] EphA4-A02-9-165 (SEQ ID
NO: 379), [0072] ECT2-A24-9-515 (SEQ ID NO: 80), [0073]
ECT2-A24-10-40 (SEQ ID NO: 100), [0074] ECT2-A24-10-101 (SEQ ID NO:
101), [0075] HIG2-A24-9-19 (SEQ ID NO: 110), [0076] HIG2-A24-9-22
(SEQ ID NO: 111), [0077] HIG2-A24-9-8 (SEQ ID NO: 387), [0078]
HIG2-A24-10-7 (SEQ ID NO: 112), [0079] HIG2-A24-10-18 (SEQ ID NO:
394), [0080] HIG2-A02-9-8 (SEQ ID NO: 114), [0081] HIG2-A02-9-15
(SEQ ID NO: 116), [0082] HIG2-A02-9-4 (SEQ ID NO: 117), [0083]
HIG2-A02-10-8 (SEQ ID NO: 121), [0084] INHBB-A24-9-180 (SEQ ID NO:
395), [0085] INHBB-A24-10-180 (SEQ ID NO: 133), [0086]
INHBB-A24-10-305 (SEQ ID NO: 135), [0087] INHBB-A24-10-7 (SEQ ID
NO: 137), [0088] INHBB-A24-10-212 (SEQ ID NO: 426), [0089]
INHBB-A02-9-213 (SEQ ID NO: 143), [0090] INHBB-A02-9-174 (SEQ ID
NO: 147), [0091] INHBB-A02-9-257 (SEQ ID NO: 148), [0092]
INHBB-A02-9-313 (SEQ ID NO: 149), [0093] INHBB-A02-9-139 (SEQ ID
NO: 150), [0094] INHBB-A02-9-8 (SEQ ID NO: 152), [0095]
INHBB-A02-9-250 (SEQ ID NO: 153), [0096] INHBB-A02-10-179 (SEQ ID
NO: 154), [0097] INHBB-A02-10-237 (SEQ ID NO: 156), [0098]
INHBB-A02-10-313 (SEQ ID NO: 160), [0099] INHBB-A02-10-173 (SEQ ID
NO: 161), [0100] INHBB-A02-10-256 (SEQ ID NO: 162), [0101]
INHBB-A02-10-162 (SEQ ID NO: 163) [0102] INHBB-A02-10-85 (SEQ ID
NO: 166). [0103] KIF20A-A24-9-305 (SEQ ID NO: 174), [0104]
KIF20A-A24-9-383 (SEQ ID NO: 178), [0105] KIF20A-A24-10-304 (SEQ ID
NO: 186), [0106] KIF20A-A24-10-66 (SEQ ID NO: 194), [0107]
KNTC2-A24-9-309 (SEQ ID NO: 196), [0108] KNTC2-A24-9-124 (SEQ ID
NO: 202), [0109] KNTC2-A24-9-154 (SEQ ID NO: 210), [0110]
KNTC2-A24-9-150 (SEQ ID NO: 213), [0111] KNTC2-A24-10-452 (SEQ ID
NO: 214), [0112] KNTC2-A24-10-227 (SEQ ID NO: 217), [0113]
KNTC2-A24-10-273 (SEQ ID NO: 223), [0114] TTK-A02-9-462 (SEQ ID NO:
227), [0115] TTK-A02-9-547 (SEQ ID NO: 228), [0116] TTK-A02-9-719
(SEQ ID NO: 233), [0117] TTK-A02-10-462 (SEQ ID NO: 254), [0118]
URLC-A02-9-206 (SEQ ID NO: 271), [0119] URLC-A02-9-212 (SEQ ID NO:
272) and [0120] URLC-A02-10-211 (SEQ ID NO: 288)
[0121] These peptides are epitope peptides of each TAA restricted
by HLA-A24 or HLA-A2 (HLA-A02). Since these antigens are
over-expressed in most cancers and are associated with tumor cell
proliferation, they find utility as immunotherapeutic targets
against cancers. Exemplary cancers include, but are not limited to,
bladder cancer, breast cancer, cervical cancer, cholangiocellular
carcinoma, CML, colorectal cancer, endometriosis, esophageal
cancer, gastric cancer, diffused type gastric cancer, liver cancer,
NSCLC, lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer,
prostate cancer, renal carcinoma, SCLC, soft tissue tumor and
testicular tumor.
[0122] Accordingly, the present invention further provides methods
of treating or preventing a disease associated with the
over-expression of CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2,
TTK and/or URLC10, e.g. cancers in a subject, such methods
including the steps of administering to the subject an immunogenic
peptide of less than about 40 amino acids, often less than about 20
amino acids, usually less than about 15 amino acids and having the
amino acid sequence of SEQ ID NOs: 19, 22, 30, 34, 344, 358, 41,
44, 46, 48, 78, 376, 379, 80, 100, 101, 110, 111, 387, 112, 394,
114, 116, 117, 121, 395, 133, 135, 137, 426, 143, 147, 148, 149,
150, 152, 153, 154, 156, 160, 161, 162, 163, 166, 174, 178, 186,
194, 196, 202, 210, 213, 214, 217, 223, 227, 228, 233, 254, 271,
272 or 288.
[0123] Alternatively, the immunogenic peptide may have an amino
acid sequence as set forth in SEQ ID NOs: 19, 22, 30, 34, 344, 358,
41, 44, 46, 48, 78, 376, 379, 80, 100, 101, 110, 111, 387, 112,
394, 114, 116, 117, 121, 395, 133, 135, 137, 426, 143, 147, 148,
149, 150, 152, 153, 154, 156, 160, 161, 162, 163, 166, 174, 178,
186, 194, 196, 202, 210, 213, 214, 217, 223, 227, 228, 233, 254,
271, 272 or 288 in which 1, 2, or several (e.g., up to 5) amino
acids are substituted, deleted or added, provided the resulting
variant peptide retains the immunogenic activity (i.e., the ability
to induce CTLs specific to cells expressing CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers).
[0124] The number of residues to be substituted, deleted, or added
is generally 5 amino acids or less, preferably 4 amino acids or
less, more preferably 3 amino acids or less, even more preferably
one or two amino acids. The cancers contemplated include, but are
not limited to, bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, CML, colorectal cancer, endometriosis,
esophageal cancer, gastric cancer, diffused type gastric cancer,
liver cancer, NSCLC, lymphoma, osteosarcoma, ovarian cancer,
pancreatic cancer, prostate cancer, renal carcinoma, SCLC, soft
tissue tumor and testicular tumor. Furthermore the present
invention provides methods for preventing post-surgery recurrence
of these diseases mentioned above.
[0125] Variant peptides (i.e., peptides having an amino acid
sequence modified by substituting, deleting, or adding one, two or
several amino acid residues to an original amino acid sequence) are
known to retain the original biological activity (Mark D F et al.,
(1984) Proc Natl Acad Sci USA 81: 5662-6.; Zoller M J and Smith M,
(1982) Nucleic Acids Res 10:6487-500.; Dalbadie-McFarland G et al.,
(1982) Proc Natl Acad Sci USA 79: 6409-13.). In the context of the
present invention, it is preferable that the amino acid
modification results in conservation of the properties of the
original amino acid side-chain (a process known as conservative
amino acid substitution). Examples of properties of amino acid side
chains include hydrophobic amino acids (A, I, L, M, F, P, W, Y, V),
hydrophilic amino acids (R, D, N, C, E, Q, G, H, K, S, T), and side
chains having the following functional groups or characteristics in
common: an aliphatic side-chain (G, A, V, L, I, P); a hydroxyl
group containing side-chain (S, T, Y); a sulfur atom containing
side-chain (C, M); a carboxylic acid and amide containing
side-chain (D, N, E, Q); a base containing side-chain (R, K, H);
and an aromatic containing side-chain (H, F, Y, W). Note, the
parenthetic letters indicate the one-letter codes of amino
acids.
[0126] In preferred embodiments, the immunogenic peptide is a
nonapeptide (9-mer) or a decapeptide (10-mer). The present
invention further provides a method of inducing anti-tumor immunity
for a disease associated with the over-expression of CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers,
in a subject, such a method including the steps of administering an
immunogenic peptide of the present invention, namely one having the
amino acid sequence of SEQ ID NOs: 19, 22, 30, 34, 344, 358, 41,
44, 46, 48, 78, 376, 379, 80, 100, 101, 110, 111, 387, 112, 394,
114, 116, 117, 121, 395, 133, 135, 137, 426, 143, 147, 148, 149,
150, 152, 153, 154, 156, 160, 161, 162, 163, 166, 174, 178, 186,
194, 196, 202, 210, 213, 214, 217, 223, 227, 228, 233, 254, 271,
272 or 288, or a variant thereof (i.e., including 1, 2, or several
(e.g., up to 5) amino acid substitutions, deletions, or additions)
to the subject in need thereof. The cancers contemplated include,
but are not limited to, bladder cancer, breast cancer, cervical
cancer, cholangiocellular carcinoma, CML, colorectal cancer,
endometriosis, esophageal cancer, gastric cancer, diffused type
gastric cancer, liver cancer, NSCLC, lymphoma, osteosarcoma,
ovarian cancer, pancreatic cancer, prostate cancer, renal
carcinoma, SCLC, soft tissue tumor and testicular tumor.
[0127] In the context of the present invention, the subject is
preferably a mammal. Exemplary mammals include, but are not limited
to, e.g., a human, non-human primate, mouse, rat, dog, cat, horse,
or cow. In the present invention, the peptide can be administered
to a subject via an in vivo or ex vivo protocol. Furthermore, the
present invention also provides use of nonapeptide or decapeptide
selected from peptides having the amino acid sequence of SEQ ID
NOs: 19, 22, 30, 34, 344, 358, 41, 44, 46, 48, 78, 376, 379, 80,
100, 101, 110, 111, 387, 112, 394, 114, 116, 117, 121, 395, 133,
135, 137, 426, 143, 147, 148, 149, 150, 152, 153, 154, 156, 160,
161, 162, 163, 166, 174, 178, 186, 194, 196, 202, 210, 213, 214,
217, 223, 227, 228, 233, 254, 271, 272 or 288 (and variants
thereof) for manufacturing an immunogenic composition for treating
or preventing a disease associated with the over-expression of
CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10,
e.g. cancers. The cancers contemplated include, but are not limited
to, bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, CML, colorectal cancer, endometriosis,
esophageal cancer, gastric cancer, diffused type gastric cancer,
liver cancer, NSCLC, lymphoma, osteosarcoma, ovarian cancer,
pancreatic cancer, prostate cancer, renal carcinoma, SCLC, soft
tissue tumor and testicular tumor.
[0128] Homology analyses of the following peptides demonstrate that
they do not have significant homology with the peptides derived
from any known human gene products. [0129] CDH3-A24-9-513 (SEQ ID
NO: 19), [0130] CDH3-A24-9-406 (SEQ ID NO: 22), [0131]
CDH3-A24-10-807 (SEQ ID NO: 30), [0132] CDH3-A24-10-332 (SEQ ID NO:
34), [0133] CDH3-A24-10-655 (SEQ ID NO: 344), [0134]
CDH3-A24-10-470 (SEQ ID NO: 358), [0135] EphA4-A24-9-453 (SEQ ID
NO: 41), [0136] EphA4-A24-9-5 (SEQ ID NO: 44), [0137]
EphA4-A24-9-869 (SEQ ID NO: 46), [0138] EphA4-A24-9-420 (SEQ ID NO:
48), [0139] EphA4-A24-10-24 (SEQ ID NO: 78), [0140] EphA4-A02-9-501
(SEQ ID NO: 376), [0141] EphA4-A02-9-165 (SEQ ID NO: 379), [0142]
ECT2-A24-9-515 (SEQ ID NO: 80), [0143] ECT2-A24-10-40 (SEQ ID NO:
100), [0144] ECT2-A24-10-101 (SEQ ID NO: 101), [0145] HIG2-A24-9-19
(SEQ ID NO: 110), [0146] HIG2-A24-9-22 (SEQ ID NO: 111), [0147]
HIG2-A24-9-8 (SEQ ID NO: 387), [0148] HIG2-A24-10-7 (SEQ ID NO:
112), [0149] HIG2-A24-10-18 (SEQ ID NO: 394), [0150] HIG2-A02-9-8
(SEQ ID NO: 114), [0151] HIG2-A02-9-15 (SEQ ID NO: 116), [0152]
HIG2-A02-9-4 (SEQ ID NO: 117), [0153] HIG2-A02-10-8 (SEQ ID NO:
121), [0154] INHBB-A24-9-180 (SEQ ID NO: 395), [0155]
INHBB-A24-10-180 (SEQ ID NO: 133), [0156] INHBB-A24-10-305 (SEQ ID
NO: 135), [0157] INHBB-A24-10-7 (SEQ ID NO: 137), [0158]
INHBB-A24-10-212 (SEQ ID NO: 426), [0159] INHBB-A02-9-213 (SEQ ID
NO: 143), [0160] INHBB-A02-9-174 (SEQ ID NO: 147), [0161]
INHBB-A02-9-257 (SEQ ID NO: 148), [0162] INHBB-A02-9-313 (SEQ ID
NO: 149), [0163] INHBB-A02-9-139 (SEQ ID NO: 150), [0164]
INHBB-A02-9-8 (SEQ ID NO: 152), [0165] INHBB-A02-9-250 (SEQ ID NO:
153), [0166] INHBB-A02-10-179 (SEQ ID NO: 154), [0167]
INHBB-A02-10-237 (SEQ ID NO: 156), [0168] INHBB-A02-10-313 (SEQ ID
NO: 160), [0169] INHBB-A02-10-173 (SEQ ID NO: 161), [0170]
INHBB-A02-10-256 (SEQ ID NO: 162), [0171] INHBB-A02-10-162 (SEQ ID
NO: 163) [0172] INHBB-A02-10-85 (SEQ ID NO: 166). [0173]
KIF20A-A24-9-305 (SEQ ID NO: 174), [0174] KIF20A-A24-9-383 (SEQ ID
NO: 178), [0175] KIF20A-A24-10-304 (SEQ ID NO: 186), [0176]
KIF20A-A24-10-66 (SEQ ID NO: 194), [0177] KNTC2-A24-9-309 (SEQ ID
NO: 196), [0178] KNTC2-A24-9-124 (SEQ ID NO: 202), [0179]
KNTC2-A24-9-154 (SEQ ID NO: 210), [0180] KNTC2-A24-9-150 (SEQ ID
NO: 213), [0181] KNTC2-A24-10-452 (SEQ ID NO: 214), [0182]
KNTC2-A24-10-227 (SEQ ID NO: 217), [0183] KNTC2-A24-10-273 (SEQ ID
NO: 223), [0184] TTK-A02-9-462 (SEQ ID NO: 227), [0185]
TTK-A02-9-547 (SEQ ID NO: 228), [0186] TTK-A02-9-719 (SEQ ID NO:
233), [0187] TTK-A02-10-462 (SEQ ID NO: 254), [0188] URLC-A02-9-206
(SEQ ID NO: 271), [0189] URLC-A02-9-212 (SEQ ID NO: 272) and [0190]
URLC-A02-10-211 (SEQ ID NO: 288)
[0191] Accordingly, the possibility of unknown or undesirable
immune responses with immunotherapy against these molecules is
significantly reduced.
[0192] Regarding HLA antigens, the data presented here demonstrate
that the uses of A-24 type or A-2 type antigens (which are said to
be highly expressed among the Japanese) are favorable for obtaining
effective results. The uses of subtypes such as A-2402 and A-0201
are even more preferable. Typically, in the clinic, the type of HLA
antigen of the patient requiring treatment is investigated in
advance, which, in turn, enables the selection of appropriate
peptides having high levels of binding affinity to the patient
antigen, or having cytotoxic T cell (CTL) inducibility by antigen
presentation. Furthermore, in order to obtain peptides having high
binding affinity and CTL inducibility, substitution, deletion, or
addition of 1, 2, or several (e.g., up to 5) amino acids may be
performed based on the amino acid sequence of the naturally
occurring CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and
URLC10 partial peptide. Herein, the term "several" means refers to
5 or less, more preferably 3 or less. Furthermore, in addition to
peptides that are naturally displayed, since the regularity of the
sequences of peptides displayed by binding to HLA antigens is
already known (Kubo R T, et al., (1994) J. Immunol., 152, 3913-24.;
Rammensee H G, et al., (1995) Immunogenetics. 41:178-228.; Kondo A,
et al., (1995) J. Immunol. 155:4307-12.), modifications based on
such regularity can be performed on the immunogenic peptides of the
invention. For example, peptides possessing high HLA-24 binding
affinity in which the second amino acid from the N terminus
substituted with phenylalanine, tyrosine, methionine, or tryptophan
may be favorably used. Likewise, peptides whose C-terminal amino
acid is substituted with phenylalanine, leucine, isoleucine,
tryptophan, or methionine may also be used favorably. On the other
hand, peptides possessing high HLA-A2 (HLA-A02) binding affinity in
which the second amino acid from the N terminus substituted with
leucine or methionine, and peptides whose C-terminal amino acid is
substituted with valine or leucine may be used favorably. The
substitution is performed not only at the terminus amino acids but
also at the position of potential TCR recognition of peptides.
Several studies have demonstrated that amino acid substitutions in
a peptide can be equal to or better than the original, for example
CAP1, p53 .sub.(264-272), Her-2/neu .sub.(369-377) or gp100
.sub.(209-217) (Zaremba et al. Cancer Res. 57, 4570-4577, 1997, T.
K. Hoffmann et al. J Immunol. (2002) Feb. 1; 168(3):1338-47., S. O.
Dionne et al. Cancer Immunol Immunotherapy. (2003) 52: 199-206 and
S. O. Dionne et al. Cancer Immunology, Immunotherapy (2004) 53,
307-314). Furthermore, 1 to 2 amino acids may be added to the N
terminus and/or C terminus of the peptide.
[0193] However, when the peptide sequence is identical to a portion
of the amino acid sequence of an endogenous or exogenous protein
having a different function, side effects such as autoimmune
disorders or allergic symptoms against specific substances may be
induced. Therefore, it is preferable to avoid the situation wherein
the immunogenic sequence matches the amino acid sequence of a known
protein. This situation may be avoided by performing a homology
search using available databases. If homology searches confirm that
peptides in which 1, 2 or several different amino acids do not
exist in nature, then the danger that modifications of the
above-mentioned amino acid sequence that, for example, increase the
binding affinity with HLA antigens, and/or increase the CTL
inducibility can be avoided.
[0194] Although peptides having high binding affinity to the HLA
antigens as described above are expected to be highly effective as
cancer vaccines, the candidate peptides, which are selected
according to the presence of high binding affinity as an indicator,
must be examined for the actual presence of CTL inducibility. CTL
inducibility may be routinely confirmed by inducing
antigen-presenting cells carrying human MHC antigens (for example,
B-lymphocytes, macrophages, and dendritic cells), or more
specifically dendritic cells derived from human peripheral blood
mononuclear leukocytes, and, after stimulation with the peptide of
interest, mixing with CD8-positive cells and measuring the
cytotoxic activity against the target cells. As the reaction
system, transgenic animals produced to express a human HLA antigen
(for example, those described in BenMohamed L, et al., (2000) Hum.
Immunol.; 61(8):764-79 Related Articles, Books, Linkout.) may be
used. For example, the target cells can be radio-labeled with
.sup.51Cr and such, and cytotoxic activity can be calculated from
radioactivity released from the target cells. Alternatively, it can
be examined by measuring IFN-gamma produced and released by CTL in
the presence of antigen-presenting cells that carry immobilized
peptides, and visualizing the inhibition zone on the media using
anti-IFN-gamma monoclonal antibodies.
[0195] As a result of examining the CTL inducibility of peptides as
described above, it was discovered that those peptides having high
binding affinity to an HLA antigen did not necessarily have high
inducibility. However, nonapeptides or decapeptides selected from
the group of peptides having the amino acid sequences indicated by
the following peptides showed particularly high CTL inducibility.
[0196] CDH3-A24-9-513 (SEQ ID NO: 19), [0197] CDH3-A24-9-406 (SEQ
ID NO: 22), [0198] CDH3-A24-10-807 (SEQ ID NO: 30), [0199]
CDH3-A24-10-332 (SEQ ID NO: 34), [0200] CDH3-A24-10-655 (SEQ ID NO:
344), [0201] CDH3-A24-10-470 (SEQ ID NO: 358), [0202]
EphA4-A24-9-453 (SEQ ID NO: 41), [0203] EphA4-A24-9-5 (SEQ ID NO:
44), [0204] EphA4-A24-9-869 (SEQ ID NO: 46), [0205] EphA4-A24-9-420
(SEQ ID NO: 48), [0206] EphA4-A24-10-24 (SEQ ID NO: 78), [0207]
EphA4-A02-9-501 (SEQ ID NO: 376), [0208] EphA4-A02-9-165 (SEQ ID
NO: 379), [0209] ECT2-A24-9-515 (SEQ ID NO: 80), [0210]
ECT2-A24-10-40 (SEQ ID NO: 100), [0211] ECT2-A24-10-101 (SEQ ID NO:
101), [0212] HIG-A24-9-19 (SEQ ID NO: 110), [0213] HIG-A24-9-22
(SEQ ID NO: 111), [0214] HIG-A24-9-8 (SEQ ID NO: 387), [0215]
HIG-A24-10-7 (SEQ ID NO: 112), [0216] HIG-A24-10-18 (SEQ ID NO:
394), [0217] HIG-A02-9-8 (SEQ ID NO: 114), [0218] HIG-A02-9-15 (SEQ
ID NO: 116), [0219] HIG-A02-9-4 (SEQ ID NO: 117), [0220]
HIG-A02-10-8 (SEQ ID NO: 121), [0221] INHBB-A24-9-180 (SEQ ID NO:
395), [0222] INHBB-A24-10-180 (SEQ ID NO: 133), [0223]
INHBB-A24-10-305 (SEQ ID NO: 135), [0224] INHBB-A24-10-7 (SEQ ID
NO: 137), [0225] INHBB-A24-10-212 (SEQ ID NO: 426), [0226]
INHBB-A02-9-213 (SEQ ID NO: 143), [0227] INHBB-A02-9-174 (SEQ ID
NO: 147), [0228] INHBB-A02-9-257 (SEQ ID NO: 148), [0229]
INHBB-A02-9-313 (SEQ ID NO: 149), [0230] INHBB-A02-9-139 (SEQ ID
NO: 150), [0231] INHBB-A02-9-8 (SEQ ID NO: 152), [0232]
INHBB-A02-10-250 (SEQ ID NO: 153), [0233] INHBB-A02-10-179 (SEQ ID
NO: 154), [0234] INHBB-A02-10-237 (SEQ ID NO: 156), [0235]
INHBB-A02-10-313 (SEQ ID NO: 160), [0236] INHBB-A02-10-173 (SEQ ID
NO: 161), [0237] INHBB-A02-10-256 (SEQ ID NO: 162), [0238]
INHBB-A02-10-162 (SEQ ID NO: 163) [0239] INHBB-A02-10-85 (SEQ ID
NO: 166). [0240] KIF20A-A24-9-305 (SEQ ID NO: 174), [0241]
KIF20A-A24-9-383 (SEQ ID NO: 178), [0242] KIF20A-A24-10-304 (SEQ ID
NO: 186), [0243] KIF20A-A24-10-66 (SEQ ID NO: 194), [0244]
KNTC2-A24-9-309 (SEQ ID NO: 196), [0245] KNTC2-A24-9-124 (SEQ ID
NO: 202), [0246] KNTC2-A24-9-154 (SEQ ID NO: 210), [0247]
KNTC2-A24-9-150 (SEQ ID NO: 213), [0248] KNTC2-A24-10-452 (SEQ ID
NO: 214), [0249] KNTC2-A24-10-227 (SEQ ID NO: 217), [0250]
KNTC2-A24-10-273 (SEQ ID NO: 223), [0251] TTK-A02-9-462 (SEQ ID NO:
227), [0252] TTK-A02-9-547 (SEQ ID NO: 228), [0253] TTK-A02-9-719
(SEQ ID NO: 233), [0254] TTK-A02-10-462 (SEQ ID NO: 254), [0255]
URLC-A02-9-206 (SEQ ID NO: 271), [0256] URLC-A02-9-212 (SEQ ID NO:
272) and [0257] URLC-A02-10-211 (SEQ ID NO: 288)
[0258] As noted above, the present invention provides peptides
having cytotoxic T cell inducibility, namely those having the amino
acid sequence of SEQ ID NOs: 19, 22, 30, 34, 344, 358, 41, 44, 46,
48, 78, 376, 379, 80, 100, 101, 110, 111, 387, 112, 394, 114, 116,
117, 121, 395, 133, 135, 137, 426, 143, 147, 148, 149, 150, 152,
153, 154, 156, 160, 161, 162, 163, 166, 174, 178, 186, 194, 196,
202, 210, 213, 214, 217, 223, 227, 228, 233, 254, 271, 272 or 288
or a variant thereof (i.e., those in which 1, 2, or several amino
acids are substituted, deleted, or added).
[0259] It is preferable that the amino acid sequences composed of 9
or 10 amino acids indicated in SEQ ID NOs: 19, 22, 30, 34, 344,
358, 41, 44, 46, 48, 78, 376, 379, 80, 100, 101, 110, 111, 387,
112, 394, 114, 116, 117, 121, 395, 133, 135, 137, 426, 143, 147,
148, 149, 150, 152, 153, 154, 156, 160, 161, 162, 163, 166, 174,
178, 186, 194, 196, 202, 210, 213, 214, 217, 223, 227, 228, 233,
254, 271, 272 or 288 or a variant thereof do not match an amino
acid sequence associated with another endogenous protein.
[0260] In particular, amino acid substitution to leucine or
methionine at the second amino acid from the N terminus, amino acid
substitution to valine or leucine at the C-terminal amino acid, and
amino acid addition of 1 to 2 amino acids at the N terminus and/or
C terminus are examples of preferred variants.
[0261] One of skill in the art will recognize that in addition to
amino acid substitutions and additions, immunologically active
fragments of the peptides may also be used in the methods of the
invention. Methods for determining active fragments are well known
in the art. CTL clones obtained by stimulation by these modified
peptides can recognize the original peptides and cause damage for
cells expressing the original peptides.
[0262] Peptides of the present invention can be prepared using well
known techniques. For example, the peptides can be prepared
synthetically, using either recombinant DNA technology or chemical
synthesis. Peptides of the present invention may be synthesized
individually or as longer polypeptides composed of two or more
peptides. The peptides of the present invention are preferably
isolated, i.e., substantially free of other naturally occurring
host cell proteins and fragments thereof.
[0263] The peptides of the present invention may contain
modifications, such as glycosylation, side chain oxidation, or
phosphorylation; so long as the modifications do not destroy the
biological activity of the peptides as described herein, namely the
ability to binding to an HLA antigen and induce CTL. Other
modifications include incorporation of D-amino acids or other amino
acid mimetics that can be used, for example, to increase the serum
half life of the peptides.
[0264] Moreover, this invention may contain a method of screening
for a peptide which 1, 2, or several amino acids are substituted,
wherein said peptide comprises an amino acid sequence selected from
the group consisting of SEQ ID NO: 19, 22, 30, 34, 344, 358, 41,
44, 46, 48, 78, 80, 100, 101, 110, 111, 387, 112, 394, 395, 133,
135, 137, 426, 174, 178, 186, 194, 196, 202, 210, 213, 214, 217 or
223, said method comprising the steps of: [0265] (a) conforming no
significant sequence homology to the entire sequence of 1, 2 or
several amino acids substitute; [0266] (b) measuring the CTL
inducibility of the candidate substitute peptide; and [0267] (c)
selecting the peptide which CTL inducibility is same to or higher
than the original peptide. Alternatively, this invention may
contain a method of screening for a peptide which 1, 2, or several
amino acids are substituted, wherein said peptide comprises an
amino acid sequence selected from the group consisting of SEQ ID
NO: 376, 379, 114, 116, 117, 121, 143, 147, 148, 149, 150, 152,
153, 154, 156, 160, 161, 162, 163, 166, 227, 228, 233, 254, 271,
272 or 288, said method comprising the steps of: [0268] (a)
conforming no significant sequence homology to the entire sequence
of 1, 2 or several amino acids substitute; [0269] (b) measuring the
CTL inducibility of the candidate substitute peptide; and [0270]
(c) selecting the peptide which CTL inducibility is same to or
higher than the original peptide.
[0271] For example, in preferred embodiments, the present invention
provides a method of identifying for a peptide having an ability to
induce CTL against cells expressing at least one tumor-associated
antigen, wherein the tumor-associated antigen is antigen selected
from the group consisting of CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and URLC10, said method comprising the steps of:
[0272] (i) providing or generating at least one candidate sequence
which consists of an amino acid sequence modified by substituting,
deleting, or adding one, two or several amino acid residues to an
original amino acid sequence, wherein the original amino acid
sequence is selected from the group consisting of SEQ ID NO: 19,
22, 30, 34, 344, 358, 41, 44, 46, 48, 78, 80, 100, 101, 110, 111,
387, 112, 394, 395, 133, 135, 137, 426, 174, 178, 186, 194, 196,
202, 210, 213, 214, 217 or 223; [0273] (ii) selecting the candidate
sequence that does not have substantial significant homology with
the peptides derived from any known human gene products other than
said tumor-associated antigens; [0274] (iii) contacting a peptide
consisting of the candidate sequence selected in step (ii) with
antigen presenting cells; [0275] (iv) contacting the antigen
presenting cells of step iii with T-cells to evaluate the ability
of the peptide to stimulate the T-cells; and [0276] (v) identifying
the peptide of which CTL inducibility is same to or higher than a
peptide consisting of the original amino acid sequence.
Alternatively, in preferred embodiments, the present invention
provides a method of identifying for a peptide having an ability to
induce CTL against cells expressing at least one tumor-associated
antigen, wherein the tumor-associated antigen is antigen selected
from the group consisting of CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and URLC10, said method comprising the steps of:
[0277] (i) providing or generating at least one candidate sequence
which consists of an amino acid sequence modified by substituting,
deleting, or adding one, two or several amino acid residues to an
original amino acid sequence, wherein the original amino acid
sequence is selected from the group consisting of SEQ ID NO: 376,
379, 114, 116, 117, 121, 143, 147, 148, 149, 150, 152, 153, 154,
156, 160, 161, 162, 163, 166, 227, 228, 233, 254, 271, 272 or 288;
[0278] (ii) selecting the candidate sequence that does not have
substantial significant homology with the peptides derived from any
known human gene products other than said tumor-associated
antigens; [0279] (iii) contacting a peptide consisting of the
candidate sequence selected in step (ii) with antigen presenting
cells; [0280] (iv) contacting the antigen presenting cells of step
(iii) with T-cells to evaluate the ability of the peptide to
stimulate the T-cells; and [0281] (v) identifying the peptide of
which CTL inducibility is same to or higher than a peptide
consisting of the original amino acid sequence.
[0282] Preferably, the amino acid is substituted for a different
amino acid in which the properties of the amino acid side-chain are
conserved (a process known as conservative amino acid
substitution). Examples of properties of amino acid side chains are
hydrophobic amino acids (A, I, L, M, F, P, W, Y, V), hydrophilic
amino acids (R, D, N, C, E, Q, G, H, K, S, T), and side chains
having the following functional groups or characteristics in
common: an aliphatic side-chain (G, A, V, L, I, P); a hydroxyl
group containing side-chain (S, T, Y); a sulfur atom containing
side-chain (C, M); a carboxylic acid and amide containing
side-chain (D, N, E, Q); a base containing side-chain (R, K, H);
and an aromatic containing side-chain (H, F, Y, W). Note, the
parenthetic letters indicate the one-letter codes of amino acids.
In the present invention, substantial significant homology is, for
example, more than 90%, preferably 95%, more preferably 99% or 100%
identity with a known human gene product to be compared.
[0283] The peptides of this invention can be prepared as a
combination, which includes two or more of peptides of the
invention, for use as a vaccine for a disease associated with the
over-expression of CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2,
TTK and/or URLC10, e.g. cancers, such a vaccine inducing CTL in
vivo. The cancers contemplated include, but are not limited to,
bladder cancer, breast cancer, cervical cancer, cholangiocellular
carcinoma, CML, colorectal cancer, endometriosis, esophageal
cancer, gastric cancer, diffused type gastric cancer, liver cancer,
NSCLC, lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer,
prostate cancer, renal carcinoma, SCLC, soft tissue tumor and
testicular tumor. The peptides may be in a cocktail or may be
conjugated to each other using standard techniques. For example,
the peptides can be expressed as a single polypeptide sequence. The
peptides in the combination may be the same or different.
[0284] By administering the peptides of this invention, the
peptides are presented at a high density on the HLA antigens of
antigen-presenting cells, which, in turn, induces CTLs that
specifically react toward the complex formed between the displayed
peptide and the HLA antigen. Alternatively, antigen-presenting
cells having immobilized the peptides of this invention on their
cell surface, obtained by removing dendritic cells from the
subjects, may be stimulated by the peptides of this invention.
Re-administration of these cells to the respective subjects induces
CTL, and, as a result, aggressiveness towards the target cells can
be increased.
[0285] More specifically, the present invention provides drugs for
treating and/or preventing proliferation, metastasis, and such of a
disease associated with the over-expression of CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers, which
include one or more of peptides of the present invention, or a
polynucleotide encoding the peptides. The peptides or
polynucleotides of the present invention find particular utility in
the treatment of a disease associating CDH3, EPHA4, ECT2, HIG2,
INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers. The cancers
contemplated include, but are not limited to, bladder cancer,
breast cancer, cervical cancer, cholangiocellular carcinoma, CML,
colorectal cancer, endometriosis, esophageal cancer, gastric
cancer, diffused type gastric cancer, liver cancer, NSCLC,
lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate
cancer, renal carcinoma, SCLC, soft tissue tumor and testicular
tumor.
[0286] The peptides of this invention can be administered to a
subject directly, as a pharmaceutical composition that has been
formulated by conventional formulation methods. In such cases, in
addition to the peptides of this invention, carriers, excipients,
and such that are ordinarily used for drugs can be included as
appropriate, without particular limitations. The immunogenic
compositions of this invention may be used for treatment and
prevention of a disease associated with the over-expression of
CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10,
e.g. cancers. The cancers contemplated include, but are not limited
to, bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, CML, colorectal cancer, endometriosis,
esophageal cancer, gastric cancer, diffused type gastric cancer,
liver cancer, NSCLC, lymphoma, osteosarcoma, ovarian cancer,
pancreatic cancer, prostate cancer, renal carcinoma, SCLC, soft
tissue tumor and testicular tumor.
[0287] The immunogenic compositions for treatment and/or prevention
of a disease associated with the over-expression of CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers,
which include as the active ingredient one or more peptides of the
present invention, can further include an adjuvant so that cellular
immunity will be established effectively. Alternatively, they may
be administered with other active ingredients, such as anti-cancer
agents.
[0288] The cancers contemplated include, but are not limited to,
bladder cancer, breast cancer, cervical cancer, cholangiocellular
carcinoma, CML, colorectal cancer, endometriosis, esophageal
cancer, gastric cancer, diffused type gastric cancer, liver cancer,
NSCLC, lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer,
prostate cancer, renal carcinoma, SCLC, soft tissue tumor and
testicular tumor. Suitable formulations include granules. Suitable
adjuvants are described in the literature (Johnson AG. (1994) Clin.
Microbiol. Rev., 7:277-89.).
[0289] Exemplary adjuvants include, but are not limited to,
aluminum phosphate, aluminum hydroxide, and alum. Furthermore,
liposome formulations, granular formulations in which the drug is
bound to few-mc m diameter beads, and formulations in which a lipid
is bound to the peptide may be conveniently used. The method of
administration may be oral, intradermal, subcutaneous, intravenous
injection, or such, and may include systemic administration or
local administration to the vicinity of the targeted tumor.
[0290] The dose of the peptide(s) of this invention can be adjusted
appropriately according to the disease to be treated, age of the
patient, weight, method of administration, and such. Though the
dosage is ordinarily 0.001 mg to 1000 mg, preferably 0.01 mg to 100
mg, more preferably 0.1 mg to 10 mg, preferably administered once
in a few days to few months, one skilled in the art can readily
select the appropriate dose and method of administration, as, the
selection and optimization of these parameters is well within
routine skill.
[0291] The present invention further provides intracellular
vesicles called exosomes, which present complexes formed between
the peptides of this invention and HLA antigens on their surface.
Exosomes can be prepared, for example, by using the methods
described in detail in Published Japanese Translation of
International Publication Nos. Hei 11-510507 and 2000-512161, and
are preferably prepared using antigen-presenting cells obtained
from subjects who are targets of treatment and/or prevention. The
exosomes of this invention can be inoculated as cancer vaccines,
similarly to the peptides of this invention.
[0292] The type of HLA antigens used must match that of the subject
requiring treatment and/or prevention. For example, in the Japanese
population, HLA-A24 or HLA-A2 (HLA-A02), particularly HLA-A2402 or
HLA-A0201, is often appropriate.
[0293] In some embodiments, the vaccine compositions of the present
invention include a component which primes cytotoxic T lymphocytes.
Lipids have been identified as agents capable of priming CTL in
vivo against viral antigens. For example, palmitic acid residues
can be attached to the epsilon- and alpha-amino groups of a lysine
residue and then linked to an immunogenic peptide of the invention.
The lipidated peptide can then be administered either directly, in
a micelle or particle, incorporated into a liposome, or emulsified
in an adjuvant. As another example of a lipid priming of CTL
responses, E. coli lipoproteins, such as
tripalmitoyl-S-glycerylcysteinlyseryl-serine (P3CSS), can be used
to prime CTL when covalently attached to an appropriate peptide
(see, e.g., Deres K, et al., (1989) Nature 342:561-4.).
[0294] The immunogenic compositions of the present invention may
also include nucleic acids encoding one or more of the immunogenic
peptides disclosed here. See, e.g., Wolff J A et al., (1990)
Science 247:1465-8; U.S. Pat. Nos. 5,580,859; 5,589,466; 5,804,566;
5,739,118; 5,736,524; 5,679,647; and WO 98/04720. Examples of
DNA-based delivery technologies include "naked DNA", facilitated
(bupivicaine, polymers, peptide-mediated) delivery, cationic lipid
complexes, and particle-mediated ("gene gun") or pressure-mediated
delivery (see, e.g., U.S. Pat. No. 5,922,687).
[0295] The immunogenic peptides of the invention can also be
expressed by viral or bacterial vectors. Examples of suitable
expression vectors include attenuated viral hosts, such as vaccinia
or fowlpox. This approach involves the use of vaccinia virus, e.g.,
as a vector to express nucleotide sequences that encode the
peptide. Upon introduction into a host, the recombinant vaccinia
virus expresses the immunogenic peptide, and thereby elicits an
immune response. Vaccinia vectors and methods useful in
immunization protocols are described in, e.g., U.S. Pat. No.
4,722,848. Another suitable vector is BCG (Bacille Calmette
Guerin). BCG vectors are described in Stover C K, et al., (1991)
Nature 351:456-60. A wide variety of other vectors useful for
therapeutic administration or immunization e.g., adeno and
adeno-associated virus vectors, retroviral vectors, Salmonella
typhi vectors, detoxified anthrax toxin vectors, and the like, are
known in the art. See, e.g., Shata M T, et al., (2000) Mol. Med.
Today 6:66-71; Shedlock D J and Weiner D B., et al., (2000) J.
Leukoc. Biol. 68:793-806; and Hipp J D, et al., (2000) In Vivo
14:571-85.
[0296] The present invention also provides methods of inducing
antigen-presenting cells using one or more peptides of this
invention. The antigen-presenting cells can be induced by inducing
dendritic cells from the peripheral blood monocytes and then
contacting (stimulating) them with one or more peptides of this
invention in vitro, ex vivo or in vivo. When peptides of the
present invention are administered to the subjects,
antigen-presenting cells that have the peptides of this invention
immobilized to them are induced in the body of the subject.
Alternatively, after immobilizing the peptides of this invention to
the antigen-presenting cells, the cells can be administered to the
subject as a vaccine. For example, the ex vivo administration may
include the steps of: [0297] a: collecting antigen-presenting cells
from a subject, and [0298] b: contacting the antigen-presenting
cells of step a with a peptide of the present invention.
[0299] Alternatively, according to the present invention, use of
the peptides of this invention for manufacturing a pharmaceutical
composition inducing antigen-presenting cells is provided. Further,
the present invention also provides the peptide of the present
invention for inducing antigen-presenting cells. The
antigen-presenting cells obtained by step b can be administered to
the subject as a vaccine.
[0300] This invention also provides a method for inducing
antigen-presenting cells having a high level of cytotoxic T cell
inducibility, in which the method includes the step of transferring
genes composed of polynucleotide(s) encoding one or more peptides
of this invention to antigen-presenting cells in vitro. The
introduced genes may be in the form of DNAs or RNAs. For the method
of introduction, without particular limitations, various methods
conventionally performed in this field, such as lipofection,
electroporation, and calcium phosphate method may be suitably used.
More specifically, transfection may be performed as described in
Reeves M E, et al., (1996) Cancer Res., 56:5672-7.; Butterfield L
H, et al., (1998) J. Immunol., 161:5607-13.; Boczkowski D, et al.,
(1996) J. Exp. Med., 184:465-72.; Published Japanese Translation of
International Publication No. 2000-509281. By transferring the gene
into antigen-presenting cells, the gene undergoes transcription,
translation, and such in the cell, and then the obtained protein is
processed by MHC Class I or Class II, and proceeds through a
presentation pathway to present partial peptides.
[0301] The present invention further provides methods for inducing
CTL using one or more peptides of this invention. When the peptides
of this invention are administered to a subject, CTL are induced in
the body of the subject, and the strength of the immune system
targeting the cells expressing CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and/or URLC10, e.g. cancer cells in the tumor
tissues is thereby enhanced.
[0302] The cancers contemplated include, but are not limited to
bladder cancer, breast cancer, cervical cancer, cholangiocellular
carcinoma, CML, colorectal cancer, endometriosis, esophageal
cancer, gastric cancer, diffused type gastric cancer, liver cancer,
NSCLC, lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer,
prostate cancer, renal carcinoma, SCLC, soft tissue tumor and
testicular tumor. Alternatively, the peptides of the present
invention may be used in the context of an ex vivo therapeutic
method, in which subject-derived antigen-presenting cells and
CD8-positive cells or peripheral blood mononuclear leukocytes are
contacted (stimulated) with one or more peptides of this invention
in vitro, and, after inducing CTL, the cells are returned to the
subject. For example, the method may include the steps of: [0303]
a: collecting antigen-presenting cells from a subject, [0304] b:
contacting the antigen-presenting cells of step a with a peptide of
the present invention, [0305] c: mixing the antigen-presenting
cells of step b with CD.sup.8+ T cells and co-culturing so as to
induce cytotoxic T-cells, and [0306] d: collecting CD.sup.8+ T
cells from the co-culture of step c.
[0307] Alternatively, according to the present invention, use of
the peptides of this invention for manufacturing a pharmaceutical
composition inducing CTLs is provided. Further, the present
invention also provides the peptide of the present invention for
inducing CTLs. The CD.sup.8+ T cells having cytotoxic activity
obtained by step d can be administered to the subject as a
vaccine.
[0308] The present invention further provides isolated cytotoxic T
cells induced using the peptides of this invention. The cytotoxic T
cells, induced by stimulation with an antigen-presenting cell
presenting one or more peptides of this invention, are preferably
derived from subjects who are the target of treatment and/or
prevention, and can be administered alone or in combination with
other drugs, including one or more peptides of this invention or
exosomes having anti-tumor activity. The obtained cytotoxic T cells
act specifically against target cells presenting the peptides of
this invention, or preferably the same peptide(s) used for
induction. The target cells may be cells that express CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10 endogenously,
or cells that are transfected with CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and/or URLC10 genes. Cells that present the
peptides of this invention on the cell surface, due to stimulation
with these peptides, can also become targets of attack.
[0309] The present invention also provides antigen-presenting cells
presenting complexes formed between HLA antigens and one or more
peptides of this invention. The antigen-presenting cells, obtained
through contact with the peptides of this invention or the
nucleotides encoding such peptides, are preferably derived from
subjects who are the target of treatment and/or prevention, and can
be administered as vaccines, alone or in combination with other
drugs, including the peptides, exosomes, or cytotoxic T cells of
the present invention.
[0310] The present invention also provides a composition composed
of nucleic acids encoding polypeptides that are capable of forming
a subunit of a T cell receptor (TCR), and methods of using the
same. The TCR subunits have the ability to form TCRs that confer
specificity to T cells for tumor cells presenting CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK or URLC10. By using the known
method in the art, the nucleic acids of alpha- and beta-chain as
the TCR subunits of the CTL induced with one or more peptides of
this invention may be identified (WO2007/032255 and Morgan et al.,
J Immunol, 171, 3288 (2003)). The derivative TCRs preferably bind
target cells displaying the CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A,
KNTC2, TTK or URLC10 peptide with high avidity, and optionally
mediate efficient killing of target cells presenting the CDH3,
EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK or URLC10 peptide in
vivo and in vitro.
[0311] The nucleic acids encoding the TCR subunits can be
incorporated into suitable vectors e.g. retroviral vectors. These
vectors are well known in the art. The nucleic acids or the vectors
containing them usefully can be transferred into a T cell, which T
cell is preferably from a patient. Advantageously, the invention
provides an off-the-shelf composition allowing rapid modification
of a patient's own T cells (or those of another mammal) to rapidly
and easily produce modified T cells having excellent cancer cell
killing properties.
[0312] Also, the present invention provides CTLs which are prepared
by transduction with the nucleic acids encoding the TCR subunits
polypeptides binding with CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A,
KNTC2, TTK or URLC10 peptide e.g. SEQ ID NOs: 19, 22, 30, 34, 344,
358, 41, 44, 46, 48, 78, 376, 379, 80, 100, 101, 110, 111, 387,
112, 394, 114, 116, 117, 121, 395, 133, 135, 137, 426, 143, 147,
148, 149, 150, 152, 153, 154, 156, 160, 161, 162, 163, 166, 174,
178, 186, 194, 196, 202, 210, 213, 214, 217, 223, 227, 228, 233,
254, 271, 272 or 288 in the context of HLA-A24 or HLA-A2 (HLA-A02).
The transduced CTLs are capable of homing to cancer cells in vivo,
and expanded by well known culturing method in vitro (e.g.,
Kawakami et al., J Immunol., 142, 3452-3461 (1989)). The T cells of
the invention can be used to form an immunogenic composition useful
in treating or preventing cancer in a patient in need of therapy or
protection (WO2006/031221).
[0313] In the context of the present invention, the term "vaccine"
(also referred to as an immunogenic composition) refers to a
substance that induces anti-tumor immunity or suppresses cancers
upon inoculation into animals. According to the present invention,
polypeptides having the amino acid sequence of SEQ ID NO: 19, 22,
30, 34, 344, 358, 41, 44, 46, 48, 78, 80, 100, 101, 110, 111, 387,
112, 394, 395, 133, 135, 137, 426, 174, 178, 186, 194, 196, 202,
210, 213, 214, 217 or 223 were suggested to be HLA-A24 restricted
epitope peptides and those of SEQ ID NO: 376, 379, 114, 116, 117,
121, 143, 147, 148, 149, 150, 152, 153, 154, 156, 160, 161, 162,
163, 166, 227, 228, 233, 254, 271, 272 or 288 were suggested to be
HLA-A2 (HLA-A02) restricted epitope peptides that may induce potent
and specific immune response against cells expressing CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancer
cells expressing CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK
and/or URLC10. The cancers contemplated include, but are not
limited to bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, CML, colorectal cancer, endometriosis,
esophageal cancer, gastric cancer, diffused type gastric cancer,
liver cancer, NSCLC, lymphoma, osteosarcoma, ovarian cancer,
pancreatic cancer, prostate cancer, renal carcinoma, SCLC, soft
tissue tumor and testicular tumor.
[0314] Thus, the present invention also encompasses a method of
inducing anti-tumor immunity using polypeptides having the amino
acid sequence of SEQ ID NO: 19, 22, 30, 34, 344, 358, 41, 44, 46,
48, 78, 376, 379, 80, 100, 101, 110, 111, 387, 112, 394, 114, 116,
117, 121, 395, 133, 135, 137, 426, 143, 147, 148, 149, 150, 152,
153, 154, 156, 160, 161, 162, 163, 166, 174, 178, 186, 194, 196,
202, 210, 213, 214, 217, 223, 227, 228, 233, 254, 271, 272 or 288
or a variant thereof (i.e., including 1, 2, or several (e.g., up to
5) amino acid substitutions, deletions, or additions). In general,
anti-tumor immunity includes immune responses such as follows:
[0315] an induction of cytotoxic lymphocytes against tumors
containing cells expressing CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A,
KNTC2, TTK and/or URLC10, [0316] an induction of antibodies that
recognize tumors containing cells expressing CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, and [0317] an
induction of anti-tumor cytokine production.
[0318] Therefore, when a certain peptide induces any one of these
immune responses upon inoculation into an animal, the peptide is
decided to have anti-tumor immunity inducing effect. The induction
of the anti-tumor immunity by a peptide can be detected by
observing in vivo or in vitro the response of the immune system in
the host against the peptide.
[0319] For example, a method for detecting the induction of
cytotoxic T lymphocytes is well known. A foreign substance that
enters the living body is presented to T cells and B cells by the
action of antigen-presenting cells (APCs). T cells that respond to
the antigen presented by APC in antigen specific manner
differentiate into cytotoxic T cells (also referred to as cytotoxic
T lymphocytes or CTLs) due to stimulation by the antigen, and then
proliferate; this process is referred to herein as "activation" of
T cells. Therefore, CTL induction by a certain peptide can be
evaluated by presenting the peptide to a T cell by APC, and
detecting the induction of CTL. Furthermore, APCs have the effect
of activating CD4+ T cells, CD8+ T cells, macrophages, eosinophils
and NK cells. Since CD4+ T cells are also important in anti-tumor
immunity, the anti-tumor immunity inducing action of the peptide
can be evaluated using the activation effect of these cells as
indicators.
[0320] A method for evaluating the inducing action of CTL using
dendritic cells (DCs) as APC is well known in the art. DC is a
representative APC having the strongest CTL inducing action among
APCs. In this method, the test polypeptide is initially contacted
with DC and then this DC is contacted with T cells. Detection of T
cells having cytotoxic effects against the cells of interest after
the contact with DC shows that the test polypeptide has an activity
of inducing the cytotoxic T cells. Activity of CTL against tumors
can be detected, for example, using the lysis of .sup.51Cr-labeled
tumor cells as the indicator. Alternatively, it is well known to
evaluate the degree of tumor cell damage using 3H-thymidine uptake
activity or LDH (lactose dehydrogenase)-release as the indicator.
Furthermore, it can be also examined by measuring IFN-gamma
produced and released by CTL in the presence of antigen-presenting
cells that carry immobilized peptides by visualizing using
anti-IFN-gamma antibodies, such as an ELISPOT assay.
[0321] Apart from DC, peripheral blood mononuclear cells (PBMCs)
may also be used as the APC. The induction of CTL is reported to be
enhanced by culturing PBMC in the presence of GM-CSF and IL-4.
Similarly, CTL has been shown to be induced by culturing PBMC in
the presence of keyhole limpet hemocyanin (KLH) and IL-7.
[0322] The test polypeptides confirmed to possess CTL inducing
activity by these methods are polypeptides having DC activation
effect and subsequent CTL inducing activity. Therefore,
polypeptides that induce CTL against cells expressed CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10 are useful as
vaccines against diseases associating CDH3, EPHA4, ECT2, HIG2,
INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers. Furthermore,
APC that have acquired the ability to induce CTL against a disease
associated with the over-expression of CDH3, EPHA4, ECT2, HIG2,
INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers, by
contacting with the polypeptides are useful as vaccines against the
disease. Furthermore, CTL that have acquired cytotoxicity due to
presentation of the polypeptide antigens by APC can be also used as
vaccines against a disease associating CDH3, EPHA4, ECT2, HIG2,
INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers. Such
therapeutic methods for a disease associating CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers, using
anti-tumor immunity due to APC and CTL, are referred to as cellular
immunotherapy. The cancers contemplated include, but are not
limited to, bladder cancer, breast cancer, cervical cancer,
cholangiocellular carcinoma, CML, colorectal cancer, endometriosis,
esophageal cancer, gastric cancer, diffused type gastric cancer,
liver cancer, NSCLC, lymphoma, osteosarcoma, ovarian cancer,
pancreatic cancer, prostate cancer, renal carcinoma, SCLC, soft
tissue tumor and testicular tumor.
[0323] Generally, when using a polypeptide for cellular
immunotherapy, efficiency of the CTL-induction can be increased by
combining a plurality of polypeptides having different structures
and contacting them with DC. Therefore, when stimulating DC with
protein fragments, it is advantageous to use a mixture of multiple
types of fragments.
[0324] The induction of anti-tumor immunity by a polypeptide can be
further confirmed by observing the induction of antibody production
against tumors. For example, when antibodies against a polypeptide
are induced in a laboratory animal immunized with the polypeptide,
and when growth, proliferation and/or metastasis of tumor cells is
suppressed by those antibodies, the polypeptide is determined to
induce anti-tumor immunity.
[0325] Anti-tumor immunity can be induced by administering a
vaccine of this invention, and the induction of anti-tumor immunity
enables treatment and prevention of a disease associated with the
over-expression of CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2,
TTK and/or URLC10, e.g. cancers. Therapy against or prevention of
the onset of a disease associated with the over-expression of CDH3,
EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g.
cancers, may include inhibition of the growth of cells expressing
CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10,
e.g. cancer cells, involution of these cells and suppression of
occurrence of these cells, e.g. cancer cells. Decrease in mortality
of individuals having a disease associating CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers,
decrease of the disease markers in the blood, alleviation of
detectable symptoms accompanying the disease and such are also
included in the therapy or prevention of the disease, e.g. cancers.
Such therapeutic and preventive effects are preferably
statistically significant, for example, observed at a significance
level of 5% or less, wherein the therapeutic or preventive effect
of a vaccine against a disease associating CDH3, EPHA4, ECT2, HIG2,
INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers, is compared
to a control without vaccine administration. For example, Student's
t-test, the Mann-Whitney U-test or ANOVA may be used for
determining statistical significance.
[0326] In that the present invention provides a method for
treating, or preventing a disease associated with the
over-expression of CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2,
TTK and/or URLC10, e.g. cancers, the therapeutic compounds or
compositions may be administered prophylactically or
therapeutically to subjects suffering from or at risk of (or
susceptible to) developing the disease. Such subjects may be
identified using standard clinical methods. In the context of the
present invention, prophylactic administration occurs prior to the
manifestation of overt clinical symptoms of disease, such that a
disease or disorder is prevented or alternatively delayed in its
progression. In the context of the field of medicine, the term
"prevent" encompasses any activity which reduces the burden of
mortality or morbidity from disease. Prevention can occur at
primary, secondary and tertiary prevention levels. While primary
prevention avoids the development of a disease, secondary and
tertiary levels of prevention encompass activities aimed at
preventing the progression of a disease and the emergence of
symptoms as well as reducing the negative impact of an already
established disease by restoring function and reducing
disease-related complications.
[0327] In the context of cancer treatment, the term "efficacious"
refers to a treatment that leads to a decrease in size, prevalence
or metastatic potential of cancer in a subject. When a treatment is
applied prophylactically, "efficacious" means that the treatment
retards or prevents occurrence of cancer or alleviates a clinical
symptom of cancer. The assessment of cancer can be made using
standard clinical protocols. Furthermore, the efficaciousness of a
treatment may be determined in association with any known method
for diagnosing or treating cancer. For example, cancer can be
diagnosed histopathologically or by identifying symptomatic
anomalies.
[0328] The above-mentioned peptide, having immunological activity,
or a polynucleotide or vector encoding such a peptide, may be
combined with an adjuvant. An adjuvant refers to a compound that
enhances the immune response against the peptide when administered
together (or successively) with the peptide having immunological
activity. Examples of suitable adjuvants include cholera toxin,
salmonella toxin, alum and such, but are not limited thereto.
Furthermore, a vaccine of this invention may be combined
appropriately with a pharmaceutically acceptable carrier. Examples
of such carriers are sterilized water, physiological saline,
phosphate buffer, culture fluid and such. Furthermore, the vaccine
may contain as necessary, stabilizers, suspensions, preservatives,
surfactants and such. The vaccine is administered systemically or
locally. Vaccine administration may be performed by single
administration or boosted by multiple administrations.
[0329] When using APC or CTL as the vaccine of this invention, a
disease associated with the over-expression of CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or URLC10, e.g. cancers, can be
treated or prevented, for example, by the ex vivo method. More
specifically, PBMCs of the subject receiving treatment or
prevention are collected, contacted ex vivo with a peptide of the
present invention. Following the induction of APC or CTL, the cells
may be administered to the subject. APC can be also induced by
introducing a vector encoding the peptide into PBMCs ex vivo. APC
or CTL induced in vitro can be cloned prior to administration. By
cloning and growing cells having high activity of damaging target
cells, cellular immunotherapy can be performed more effectively.
Furthermore, APC and CTL isolated in this manner may be used for
cellular immunotherapy not only against individuals from whom the
cells are derived, but also against similar types of diseases in
other individuals.
[0330] Aspects of the present invention are described in the
following examples, which are presented only to illustrate the
present invention and to assist one of ordinary skill in making and
using the same. The examples are not intended in any way to
otherwise limit the scope of the invention.
[0331] Although methods and materials similar or equivalent to
those described herein can be used in the practice or testing of
the present invention, suitable methods and materials are described
below.
EXAMPLES
[0332] Hereinafter, the present invention is exemplified, but not
restricted, by the following Examples. However, materials, methods
and such described herein only illustrate aspects of the invention
and in no way are intended to limit the scope of the present
invention. As such, materials, methods and such similar or
equivalent to those described therein may be used in the practice
or testing of the present invention.
Example 1
Materials and Methods
Cell Lines
[0333] A24-LCL cells (HLA-A24), human B-lymphoblastoid cell line,
was established by transforming with Epstain-barr virus. T2 cell,
COS7, A498, Caki-2 and HEK 293 were purchased from ATCC. Caki-1 and
MIAPaca-2 were purchased from JCRB. PK-45P, PK-59, TE-5 and TE-6
were purchased from TKG. 293 T was purchased from GenHunter.
Candidate Selection of Peptide Derived from CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and URLC10
[0334] 9-mer and 10-mer peptides derived from CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK or URLC10 that bind to HLA-A*2402
or HLA-A*0201 molecule were predicted using the binding prediction
software "BIMAS"
(http://bimas.dcrt.nih.gov/cgi-bin/molbio/ken_parker_comboform)
(Parker K C, et al., (1994) J Immunol.; 152(1):163-75.; Kuzushima
K, et al., (2001) Blood.; 98(6):1872-81.). These peptides were
synthesized by Sigma (Sapporo, Japan) according to the standard
solid phase synthesis method and purified by reversed phase HPLC.
The purity (>90%) and the identity of the peptides were
determined by analytical HPLC and mass spectrometry analysis,
respectively. Peptides were dissolved in dimethylsulfoxide (DMSO)
at 20 mg/ml and stored at -80 degrees C.
In Vitro CTL Induction
[0335] Monocyte-derived dendritic cells (DCs) were used as
antigen-presenting cells (APCs) to induce CTL responses against
peptides presented on HLA. DCs were generated in vitro as described
elsewhere (Nukaya I et al., (1999) Int. J. Cancer 80, 92-7., Tsai V
et al., (1997) J. Immunol 158:1796-802.). Briefly, peripheral blood
mononuclear cells (PBMCs) isolated from a normal volunteer
(HLA-A*2402 and/or HLA-A*0201) by Ficoll-Paque (Pharmacia) solution
were separated by adherence to a plastic tissue culture flask
(Becton Dickinson) so as to enrich them for the monocyte fraction.
The monocyte-enriched population was cultured in the presence of
1000 U/ml of GM-CSF (Genzyme) and 1000 U/ml of IL-4 (Genzyme) in
AIM-V(Invitrogen) containing 2% heat-inactivated autologous serum
(AS). After 7 days in the culture, the cytokine-generated DCs were
pulsed with 20 micro g/ml of the synthesized peptides in the
presence of 3 micro g/ml of beta 2-microglobulin for 4 hrs at 20
degrees C. in AIM-V. These peptide-pulsed DCs were then inactivated
by MMC (30 micro g/ml for 30 mins) and mixed at a 1:20 ratio with
autologous CD8.sup.+ T cells, obtained by positive selection with
Dynabeads M-450 CD8 (Dynal) and DETACHa BEAD.TM. (Dynal). These
cultures were set up in 48-well plates (Corning); each well
contained 1.5.times.10.sup.4 peptide-pulsed DCs, 3.times.10.sup.5
CD8.sup.+ T cells and 10 ng/ml of IL-7 (Genzyme) in 0.5 ml of
AIM-V/2% AS. Three days later, these cultures were supplemented
with IL-2 (CHIRON) to a final concentration of 20 IU/ml. On day 7
and 14, the T cells were further restimulated with the autologous
peptide-pulsed DCs. The DCs were prepared each time by the same way
described above. CTL was tested against peptide-pulsed A24-LCL
cells or T2 cells after the 3rd round of peptide stimulation on day
21.
CTL Expansion Procedure
[0336] CTLs were expanded in culture using the method similar to
that described by Riddell S R, et al., (Walter E A et al., (1995) N
Engl J Med 333:1038-44.; Riddel S R, et al., (1996) Nature Med.
2:216-23.). A total 5.times.10.sup.4 of CTLs were resuspended in 25
ml of AIM-V/5% AS with 2 kinds of human B-lymphoblastoid cell
lines, inactivated by MMC, in the presence of 40 ng/ml of anti-CD3
monoclonal antibody (Pharmingen). One day after initiating the
cultures, 120 IU/ml of IL-2 were added to the cultures. The
cultures were fed with fresh AIM-V/5% AS containing 30 IU/ml of
IL-2 on days 5, 8 and 11.
Establishment of CTL Clones
[0337] The dilutions were made to have 0.3, 1, and 3 CTLs/well in
96 round-bottomed micro titer plate (Nalge Nunc International).
CTLs were cultured with 7.times.10.sup.4 cells/well of 2 kinds of
human B-lymphoblastoid cell lines, 30 ng/ml of anti-CD3 antibody,
and 125 U/ml of IL-2 in total of 150 micro 1/well of AIM-V
containing 5% AS. 50 micro 1 /well of IL-2 was added to the medium
10 days later so that IL-2 became 125 U/ml in the final
concentration. CTL activity of CTLs was tested on the 14th day, and
CTL clones were expanded using the same method above.
Specific CTL Activity
[0338] To examine the specific CTL activity, IFN-gamma ELISPOT
assay and IFN-gamma ELISA assay were performed. Briefly,
peptide-pulsed A24-LCL or T2 cell (1.times.10.sup.4/well) was
prepared as stimulator cells. Cultured Cells in 48 wells or CTL
clones after limiting dilution were used as responder cells.
IFN-gamma ELISPOT assay and ELISA assay were performed under
manufacture procedure.
Establishment of the Cells Forcibly Expressing Either or Both of
the Target Gene and HLA-A02 or HLA-A24
[0339] The cDNA encoding an open reading frame of taget genes or
HLA-A02 or HLA-A24 was amplified by PCR. The PCR-amplified product
was cloned into pcDNA3.1 myc-His vector (Invitrogen). The plasmids
were transfected into the taget cells, HLA-A02 and HLA-A24-null
normal human cell line COS7 or 293T using lipofectamine
(Invitrogen) according to the manufacturer's recommended
procedures. Alternatively, the plasmid contained the target genes
were transfected into A24-LCL by electroporation using GenePulserII
(Biorad). Briefly, 2.5.times.10.sup.6 A24-LCL cells were pulsed
with 10 mcg prasmid at 140V and 1000 micro F. After 2 days from
transfection, the transfected cells were treated with Cell
dissociation solution and used as the target cells for CTL activity
assay.
Cytotoxicity Assay
[0340] Cytotoxic activity was evaluated by a four-hour .sup.51Cr
release assay. The target cells were pulsed with a 20 micro g/mL
concentration of peptide overnight. The target cells were labeled
with 100 micro Ci of Na.sub.2.sup.51CrO.sub.4 at 37 degrees C. for
one hour, and then washed three times with RPMI1640. The target
cells (1.times.10.sup.4/100 micro L) and 100 micro L of effector
cells at various numbers with a total volume of 200 micro L were
placed into a round-bottomed 96-well microtiter plate (Corning),
and cultured at 37 degrees C. in a CO.sub.2 incubator for four
hours. After culturing, 100 micro L of the supernatant was
collected from each well, and measured the radioactivity using a
gamma counter. Spontaneous release was the radioactivity from the
target cells with medium in the absence of effector cells, and
maximum release was the radioactivity from the target cells with 1
M HCl.
[0341] The Percentage of specific cytotoxicity was determined by
calculating as following formula:
% Specific lysis=[(experimental release-spontaneous
release)/(maximum release-spontaneous release)].times.100.
Results
Enhanced CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and
URLC10 Expression in Cancers
[0342] The global gene expression profile data obtained from
various cancers using cDNA-microarray revealed that the expression
of the following genes was elevated. [0343] CDH3 (GenBank Accession
No. NM_001793; SEQ ID Nos.1, 2), [0344] EPHA4 (GenBank Accession
No. L36645; SEQ ID Nos.3, 4), [0345] ECT2 (GenBank Accession No.
AY376439; SEQ ID Nos.5, 6), [0346] HIG2 (GenBank Accession No.
NM_013332; SEQ ID Nos.7, 8), [0347] INHBB (GenBank Accession No.
NM_002193; SEQ ID Nos.9, 435, 10, 436), [0348] KIF20A (GenBank
Accession No. NM_005733; SEQ ID Nos.11, 12), [0349] KNTC2 (GenBank
Accession No. AF017790; SEQ ID Nos.13, 14), [0350] TTK (GenBank
Accession No. NM_003318; SEQ ID Nos.15, 16) and [0351] URLC10
(GenBank Accession No. NM_017527; SEQ ID Nos.17, 18) [0352] CDH3
expression was validly elevated in the following cancers in
comparison with corresponding normal tissue. [0353] 26 out of 34
bladder cancer, [0354] 17 out of 19 cervical cancer, [0355] 19 out
of 19 cholangiocellular carcinoma, [0356] 30 out of 34 colorectal
cancer, [0357] 20 out of 21 endometriosis, [0358] 13 out of 20
gastric cancer, [0359] 7 out of 8 diffuse-type gastric cancer,
[0360] 36 out of 37 NSCLC, [0361] 16 out of 16 pancreatic cancer,
[0362] 21 out of 21 soft tissue tumor and [0363] 10 out of 10
testicular tumor
[0364] EPHA4 expression was validly elevated in the following
cancers in comparison with corresponding normal tissue. [0365] 14
out of 34 bladder cancer, [0366] 8 out of 14 cervical cancer,
[0367] 10 out of 25 cholangiocellular carcinoma, [0368] 5 out of 15
endometriosis, [0369] 5 out of 8 diffuse-type gastric cancer,
[0370] 5 out of 5 ovarian cancer, [0371] 14 out of 14 pancreatic
cancer, 20 out of 51 prostate cancer and 14 out of 23 soft tissue
tumor
[0372] ECT2 expression was validly elevated in the following
cancers in comparing with corresponding normal tissue. [0373] 17
out of 19 bladder cancer, [0374] 5 out of 12 breast cancer, [0375]
14 out of 14 cervical cancer, [0376] 13 out of 13 cholangiocellular
carcinoma, [0377] 5 out of 5 CML, [0378] 7 out of 8 colorectal
cancer, [0379] 12 out of 16 esophageal cancer, [0380] 6 out of 16
NSCLC, [0381] 8 out of 10 lymphoma, [0382] 1 out of 1 pancreatic
cancer, [0383] 10 out of 13 prostate cancer, [0384] 3 out of 6
renal carcinoma and [0385] 12 out of 13 SCLC cancer [0386] HIG2
expression was validly elevated in 19 out of 20 renal cancer and 7
out of 9 soft tissue tumor in comparing with corresponding normal
tissue. [0387] INHBB expression was validly elevated in the
following cancers in comparing with corresponding normal tissue.
[0388] 10 out of 21 cholangiocellular carcinoma, [0389] 12 out of
12 esophageal cancer, [0390] 10 out of 13 NSCLC, [0391] 22 out of
24 renal carcinoma, [0392] 8 out of 14 SCLC cancer and [0393] 45
out of 49 soft tissue tumor
[0394] KIF20A expression was validly elevated in the following
cancers in comparing with corresponding normal tissue. [0395] 31
out of 31 bladder cancer, [0396] 38 out of 61 breast cancer, [0397]
10 out of 11 cholangiocellular carcinoma, [0398] 7 out of 19
esophageal cancer, [0399] 21 out of 22 NSCLC, [0400] 6 out of 6
ovarian cancer, [0401] 17 out of 36 prostate cancer, [0402] 6 out
of 11 renal carcinoma and [0403] 15 out of 15 SCLC
[0404] KNTC2 expression was validly elevated in the following
cancers in comparing with corresponding normal tissue. [0405] 30
out of 32 bladder cancer, [0406] 47 out of 56 breast cancer, [0407]
10 out of 10 cervical cancer, [0408] 16 out of 22 cholangiocellular
carcinoma, [0409] 17 out of 37 CML, [0410] 3 out of 10 colorectal
cancer, [0411] 11 out of 46 esophagus cancer, [0412] 15 out of 19
NSCLC, [0413] 7 out of 8 lymphoma, [0414] 20 out of 24
osteosarcoma, [0415] 3 out of 5 ovarian cancer, [0416] 2 out of 2
pancreatic cancer, [0417] 15 out of 37 prostate cancer, [0418] 14
out of 19 renal carcinoma, [0419] 15 out of 15 SCLC and [0420] 40
out of 59 soft tissue tumor
[0421] TTK expression was validly elevated in the following cancers
in comparing with corresponding normal tissue. [0422] 27 out of 27
bladder cancer, [0423] 25 out of 30 breast cancer, [0424] 15 out of
16 cervical cancer, [0425] 10 out of 10 cholangiocellular
carcinoma, [0426] 5 out of 7 CML, [0427] 6 out of 10 colorectal
cancer, [0428] 24 out of 44 esophageal cancer, [0429] 8 out of 15
liver cancer, [0430] 12 out of 12 NSCLC, [0431] 6 out of 6
lymphoma, [0432] 13 out of 16 osteoblastoma, [0433] 12 out of 17
prostate cancer, [0434] 15 out of 15 SCLC and [0435] 16 out of 33
soft tissue tumor
[0436] URLC10 expression was validly elevated in the following
cancers in comparing with corresponding normal tissue [0437] 29 out
of 29 bladder cancer, [0438] 15 out of 16 cervical cancer, [0439] 7
out of 7 cholangiocellular carcinoma, [0440] 7 out of 19 esophageal
cancer, [0441] 3 out of 3 gastric cancer, 24 out of 27 NSCLC,
[0442] 15 out of 19 osteosarcoma, [0443] 4 out of 5 pancreatic
cancer, [0444] 33 out of 43 soft tissue tumor.
TABLE-US-00001 [0444] TABLE 1 Ratio of cases observed up-regulation
of CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK or URLC10 in
cancerous tissue as compared to normal corresponding tissue CDH3
EPHA4 ECT2 HIG2 INHBB Bladder cancer 26/34 14/34 17/19 -- -- Breast
cancer -- -- 5/12 -- -- Cervical cancer 17/19 8/14 14/14 -- --
Cholangiocellularcarcinoma 19/19 10/25 13/13 -- 10/21 CML -- -- 5/5
-- -- Colectal cancer 30/34 -- 7/8 -- -- Endometriosis 20/21 5/15
-- -- -- Esophageal cancer -- -- 12/16 -- 12/12 Gastric camcer
13/20 -- -- -- -- Diffuse-type Gastric cancer 7/8 5/8 -- -- --
Liver cancer -- -- -- -- -- non-small cell lung cancer 36/37 --
6/16 -- 10/13 Lymphoma -- -- 8/10 -- -- Osteosarcoma -- -- -- -- --
Ovarian cancer -- 5/5 -- -- -- Pancreatic cancer 16/16 14/14 1/1 --
-- Prostate cancer -- 20/51 10/13 -- -- Renal carcinoma -- -- 3/6
19/20 22/24 Small cell lung cancer -- -- 12/13 -- 8/14 Soft tissue
tumor 21/21 14/23 -- 7/9 45/49 Testicular tumor 10/10 -- -- -- --
KIF20A KNTC2 TTK URLC10 Bladder cancer 31/31 30/32 27/27 29/29
Breast cancer 38/61 47/56 25/30 -- Cervical cancer -- 10/10 15/16
15/16 Cholangiocellularcarcinoma 10/11 16/22 10/10 7/7 CML -- 17/37
5/7 -- Colectal cancer -- 3/10 6/10 -- Endometriosis -- -- -- --
Esophageal cancer 7/19 11/46 24/44 7/19 Gastric camcer -- -- -- 3/3
Diffuse-type Gastric cancer -- -- -- -- Liver cancer -- -- 8/15 --
non-small cell lung cancer 21/22 15/19 12/12 24/27 Lymphoma -- 7/8
6/6 -- Osteosarcoma -- 20/24 13/16 15/19 Ovarian cancer -- 3/5 --
-- Pancreatic cancer 6/6 2/2 -- 4/5 Prostate cancer 17/36 15/37
12/17 -- Renal carcinoma 6/11 14/19 -- -- Small cell lung cancer
15/15 15/15 15/15 -- Soft tissue tumor -- 40/59 16/33 33/43
Testicular tumor -- -- -- --
Prediction of HLA-A24 or HLA-A2 (HLA-A02) Binding Peptides Derived
from CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK or
URLC10
[0445] Table 2 sets forth the HLA-A*2402 binding peptides for CDH3
in order of binding affinity. Table 2A sets forth 9-mer peptides
derived from CDH3 and Table 2B sets forth 10-mer peptides derived
from CDH3.
[0446] Table 3 sets forth the HLA-A*2402 and HLA-A*0201 binding
peptides for EPHA4 in order of binding affinity. Table 3A sets
forth the HLA-A*2402 binding 9-mer peptides derived from EPHA4,
Table 3B shows the HLA-A*2402 binding 10-mer peptides derived from
EPHA4 and Table 3C sets forth the HLA-A*0201 binding 9-mer peptides
derived from EPHA4.
[0447] Table 4 sets forth the HLA-A*2402 binding peptides for ECT2
in order of binding affinity. Table 4A sets forth 9-mer peptides
derived from ECT2 and Table 4B shows 10-mer peptides derived from
ECT2.
[0448] Table 5 sets forth the HLA-A*2402 and HLA-A*0201 binding
peptides for HIG2, Table 5A sets forth the HLA-A*2402 binding 9-mer
peptides derived from HIG2, Table 5B sets forth the HLA-A*2402
binding 10-mer peptides derived from HIG2, Table 5C sets forth the
HLA-A*0201 binding 9-mer peptides derived from HIG2, and Table 5D
sets forth HLA-A*0201 binding 10-mer peptides derived from
HIG2.
[0449] Table 6 sets forth the HLA-A*2402 and HLA-A*0201 binding
peptides for INHBB, Table 6A shows the HLA-A*2402 binding 9-mer
peptides derived from INHBB, Table 6B sets forth the HLA-A*2402
binding 10-mer peptides derived from INHBB, Table 6C sets forth the
HLA-A*0201 binding 9-mer peptides derived from INHBB, and Table 6D
sets forth HLA-A*0201 binding 10-mer peptides derived from
INHBB.
[0450] Table 7 sets forth the HLA-A*2402 binding peptides for
KIF20A in order of binding affinity. Table 7A sets forth 9-mer
peptides derived from KIF20A and Table 7B sets forth 10-mer
peptides derived from KIF20A.
[0451] Table 8 sets forth the HLA-A*2402 binding peptides for KNTC2
in order of binding affinity. Table 8A sets forth 9-mer peptides
derived from KNTC2 and Table 8B sets forth 10-mer peptides derived
from KNTC2.
[0452] Table 9 sets forth the HLA-A*0201 binding peptides for TTK
in order of binding affinity. Table 9A sets forth 9-mer peptides
derived from TTK and Table 9B sets forth 10-mer peptides derived
from TTK.
[0453] Table 10 sets forth the HLA-A*0201 binding peptides for
URLC10 in order of binding affinity. Table 10A sets forth 9-mer
peptides derived from URLC10 and Table 10B sets forth 10-mer
peptides derived from URLC10.
[0454] Explanation and definition about the terms in tables
Start position indicates the number of amino acid from N-terminal.
Binding score is derived from "BIMAS" described in Materials and
Methods. Positive donor number indicates the number of donors whose
CD8+-T-cells can be induced to the specific CTL by the ex vivo
stimulation with antigen-presenting cells. This is shown as
(positive donor number/whole donor number). Positive well number
indicates the number of wells where specific IFN-gamma production
can be detected by IFN-gamma ELISPOT assay. 4 to 8 wells can be
prepared from one donor. This is shown as (positive wells
number/the number of whole wells tested by IFN-gamma ELISPOT
assay). Positive CTL line indicates the number of CTL line
established from positive wells. The generation of CTL line is
determined by ELISA. This is shown as (established CTL line
number/the number of positive wells tested by IFN-gamma ELISPOT
assay). No positive donor is not defined by no detectable positive
wells, but by no established CTL line. The peptides showed by bold
character in tables possesses the stimulation activity of the T
cells. No data at positive donor number, positive well number and
positive CTL line indicating "-" means that the peptides can't be
synthesized for any reason.
TABLE-US-00002 TABLE 2A HLA-A*2402 binding 9-mer peptides derived
from CDH3 Positive Positive Strat Amino acid Binding donor well
Positive SEQ ID position sequence Score number number CTL line NO.
513 IYEVMVLAM 37.5 1/3 19 667 LFLLLVLLL 36 -- -- -- 20 30 VFREAEVTL
24 0/3 1/22 0/1 21 406 LYVEVTNEA 16.632 1/3 22 332 KYEAHVPEN 16.5
0/3 1/22 0/1 23 180 KYELFGHAV 15 0/3 1/22 0/1 24 85 RSLKERNPL 14.4
0/3 1/22 0/1 25 5 RGPLASLLL 12 0/3 2/22 0/2 26 652 KGGFILPVL 11.2
0/3 0/22 -- 27 248 TYNGVVAYS 10.5 0/3 2/22 0/2 28 65 LFSTDNDDF 10
0/3 0/22 -- 29 94 KIFPSKRIL 9.6 0/1 0/8 -- 306 221 RGSVLEGVL 9.6
0/1 0/8 -- 307 668 FLLLVLLLL 8.4 -- -- -- 308 754 IGNFIIENL 8.4 --
-- -- 309 311 TAVAVVEIL 8.4 0/1 0/8 -- 310 557 NQSPVRQVL 8.064 0/1
0/8 -- 311 611 KQDTYDVHL 8 0/1 0/8 -- 312 781 DYEGSGSDA 7.5 0/1 0/8
-- 313 165 GWLLLNKPL 7.2 0/1 0/8 -- 314 656 ILPVLGAVL 7.2 0/1 0/8
-- 315 770 TAPPYDTLL 7.2 0/1 0/8 -- 316 602 VVLSLKKFL 7.2 0/1 0/8
-- 317 665 ALLFLLLVL 7.2 -- -- -- 318 410 VTNEAPFVL 7.2 0/1 0/8 --
319 662 AVLALLFLL 7.2 -- -- -- 320 613 DTYDVHLSL 6.72 0/1 0/8 --
321 6 GPLASLLLL 6 0/1 0/8 -- 322 564 VLNITDKDL 6 0/1 0/8 -- 323 159
AVEKETGWL 6 0/1 0/8 -- 324 511 NNIYEVMVL 6 0/1 0/8 -- 325 11
LLLLQVCWL 6 -- -- -- 326 57 GCPGQEPAL 6 0/1 0/8 -- 327 293
EYTLTIQAT 6 0/1 0/8 -- 328 79 ETVQERRSL 6 0/1 0/8 -- 329 475
SYRILRDPA 6 0/1 0/8 -- 330 493 GQVTAVGTL 6 0/1 0/8 -- 331 661
GAVLALLFL 6 0/1 0/8 -- 332 388 GILTTRKGL 6 0/1 0/8 -- 333 382
HPESNQGIL 6 0/1 0/8 -- 334 663 VLALLFLLL 5.76 -- -- -- 335 598
EGDTVVLSL 5.6 0/1 0/8 -- 336 278 TISVISSGL 5.6 0/1 2/8 0/2 337 659
VLGAVLALL 5.6 0/1 0/8 -- 338 811 EWGSRFKKL 5.28 0/1 0/8 -- 339 445
KVVEVQEGI 5.04 0/1 0/8 -- 340 614 TYDVHLSLS 5 0/1 0/8 -- 341 142
FYSITGPGA 5 0/1 0/8 -- 342 246 IYTYNGVVA 5 0/1 0/8 -- 343
TABLE-US-00003 TABLE 2B HLA-A*2402 binding 10-mer peptides derived
from CDH3 positive positive strat Binding donor well positive SEQ
ID position sequence Score number number CTL line NO 807 DYLNeWGSRF
150 1/3 30 248 TYNGvVAYSI 105 0/3 4/22 0/4 31 667 LFLLlVLLLL 42 --
-- -- 32 397 DFEAkNQHTL 30 0/3 2/22 0/2 33 332 KYEAhVPENA 21 1/3 34
180 KYELFGHAVS 15 0/3 2/22 0/2 35 510 RNNIYEVMVL 12 0/3 4/22 0/4 36
5 RGPLASLLLL 12 0/3 1/22 0/1 37 477 RILRDPAGWL 12 0/3 1/22 0/1 38
556 CNQSPVRQVL 10.08 0/3 2/22 0/2 39 655 FILPvLGAVL 8.64 1/3 344
662 AVLAlLFLLL 8.64 -- -- -- 345 277 GTISvISSGL 8.4 0/3 0/20 -- 346
781 DYEGsGSDAA 7.5 0/3 0/20 -- 347 601 TVVLsLKKFL 7.2 0/3 3/20 0/3
348 158 FAVEkETGWL 7.2 0/3 0/20 -- 349 665 ALLFILLVLL 7.2 -- -- --
350 259 SQEPkDPHDL 7.2 0/3 0/20 -- 351 664 LALLfLLLVL 7.2 -- -- --
352 42 GAEQePGQAL 7.2 0/3 1/20 0/1 353 661 GAVLaLLFLL 7.2 -- -- --
354 595 VNEEgDTVVL 7.2 0/2 0/12 -- 355 340 NAVGhEVQRL 7.2 0/2 0/12
-- 356 411 TNEApFVLKL 6.6 0/2 0/12 -- 357 470 ENQKiSYRIL 6 1/2 358
10 SLLLlQVCWL 6 0/2 1/12 0/1 359 721 GLEArPEVVL 6 0/2 2/12 0/2 360
345 EVQRlTVTDL 6 0/2 4/12 0/4 361 2 GLPRgPLASL 6 0/2 3/12 0/3 362
657 LPVLgAVLAL 6 -- -- -- 363 563 QVLNiTDKDL 6 0/2 1/12 0/1 364 159
AVEKeTGWLL 6 0/2 2/12 0/2 365 492 SGQVtAVGTL 6 0/2 -- -- 366 387
QGILtTRKGL 6 0/2 -- -- 367 525 SPPTtGTGTL 6 0/2 2/12 0/2 368 358
NSPAwRATYL 6 0/2 2/12 0/2 369 122 GPFPqRLNQL 5.76 0/2 3/12 0/3 370
753 EIGNfIIENL 5.6 0/2 1/12 0/1 371 310 TTAVaVVEIL 5.6 -- -- -- 372
246 IYTYnGVVAY 5 0/2 2/12 0/2 373 805 DYDYlNEWGS 5 0/2 0/12 --
374
TABLE-US-00004 TABLE 3A HLA-A*2402 binding 9-mer peptides derived
from EPHA4 positive positive strat Binding donor well positive SEQ
ID position sequence Score number number CTL line NO 97 VYIEIKFTL
504 0/2 1/16 0/1 40 453 RYSVALAWL 400 2/3 41 25 VYPANEVTL 300 0/3
0/22 -- 42 384 HYTPQQNGL 288 0/3 1/22 0/1 43 5 FYFALFSCL 288 1/2 44
519 GYGDFSEPL 240 0/3 3/22 0/3 45 869 KFGQIVNML 67.2 1/3 46 777
AYTTRGGKI 55 0/3 1/22 0/1 47 420 KYNPNPDQS 18 1/3 48 749 RNILVNSNL
16.8 0/3 1/22 0/1 49 734 KYLSDMSYV 15 0/3 0/22 -- 50 879 KLIRNPNSL
14.4 0/3 0/22 -- 51 926 RYKDNFTAA 14.4 0/3 0/22 -- 52 834 KAIEEGYRL
14.4 0/3 0/22 -- 53 574 KYSKAKQEA 13.2 0/3 0/22 -- 54 184 AFQDVGACI
12.6 0/3 1/22 0/1 55 252 WLVPIGNCL 12.096 0/3 0/22 -- 56 326
RPPSAPLNL 12 0/3 0/22 -- 57 203 KCPLTVRNL 12 0/3 0/22 -- 58 360
SYNVVCKKC 11.55 0/3 0/22 -- 59
TABLE-US-00005 TABLE 3B HLA-A*2402 binding 10-mer peptides derived
from EPHA4 positive positive strat Binding donor well positive SEQ
ID position sequence Score number number CTL line NO 25 VYPANEVTLL
300 0/3 0/22 -- 60 244 MYCGADGEWL 200 0/3 1/22 0/1 61 657
GYTDKQRRDF 120 0/3 1/22 0/1 62 5 FYFAlFSCLF 100 -- -- -- 63 102
KFTLRDCNSL 48 0/3 1/22 0/1 64 818 SYGERPYWDM 30 0/3 2/22 0/2 65 4
IFYFALFSCL 28.8 -- -- -- 66 808 SYGIVMWEVM 25 -- -- -- 67 630
EFGEVCSGRL 24 0/3 0/22 -- 68 420 KYNPNPDQSV 21.6 0/3 0/22 -- 69 930
NFTAAGYTTL 20 0/2 0/16 -- 70 675 QFDHPNIIHL 20 0/3 0/22 -- 71 708
AFLRKNDGRF 15 0/3 0/22 -- 72 579 KQEADEEKHL 12 0/3 1/22 0/1 73 727
RGIGSGMKYL 12 0/3 0/22 -- 74 96 RVYIEIKFTL 11.2 0/2 1/16 0/1 75 507
SYVFHVRART 10.5 0/3 1/22 0/1 76 251 EWLVPIGNCL 10.08 0/3 0/22 -- 77
24 RVYPANEVTL 9.6 1/3 78 699 EYMENGSLDA 9 0/3 0/22 -- 79
TABLE-US-00006 TABLE 3C HLA-A*0201 binding 9-mer peptides derived
from EPHA4 positive positive strat Binding donor well positive SEQ
ID position sequence Score number number CTL line NO 8 ALFSCLFGI
514.942 -- -- -- 375 501 GLNPLTSYV 382.536 1/1 376 12 CLFGICDAV
126.098 0/1 1/5 0/1 377 977 QMHGRMVPV 115.534 0/1 1/5 0/1 378 165
KLNTEIRDV 111.979 1/1 379 252 WLVPIGNCL 98.267 0/1 1/5 0/1 380 879
KLIRNPNSL 74.768 0/1 1/5 0/1 381 559 VVILIAAFV 56.902 -- -- -- 382
812 VMWEVMSYG 39.386 0/1 0/5 -- 383 728 GIGSGMKYL 37.157 0/1 0/5 --
384 750 NILVNSNLV 35.385 0/1 1/5 0/1 385 937 TTLEAVVHV 33.705 0/1
1/5 0/1 386
TABLE-US-00007 TABLE 4A HLA-A*2402 binding 9-mer peptides derived
from ECT2 positive positive strat Binding donor well positive SEQ
ID position sequence Score number number CTL line NO 515 TYPPFVNFF
216 1/1 80 140 LYCTSMMNL 200 0/1 0/8 -- 81 298 LYVVKQEWF 150 0/1
0/8 -- 82 435 NYVNILATI 105 0/1 0/8 -- 83 773 IYTADPESF 100 0/1 0/8
-- 84 110 LYKADCRVI 50 0/1 0/8 -- 85 739 SFQMTSDEL 33 0/1 0/8 -- 86
504 IFLKYSKDL 30 0/1 0/8 -- 87 867 FFERRSHTL 30 0/1 0/8 -- 88 178
DFNSKVTHL 30 0/1 0/8 -- 89 61 KQEELIKAL 17.28 0/1 0/8 -- 90 657
RGEQVTLFL 16.8 0/1 2/8 0/2 91 568 RLPSVALLL 16.8 0/1 0/8 -- 92 550
KPECGRQSL 14.4 0/1 0/8 -- 93 470 IFGSIPDIF 14 0/1 0/8 -- 94 116
RVIGPPVVL 12 0/1 0/8 -- 95 507 KYSKDLVKT 11 0/1 0/8 -- 96 223
DFYAAVDDF 10 0/1 0/8 -- 97
TABLE-US-00008 TABLE 4B HLA-A*2402 binding 10-mer peptides derived
from ECT2 positive positive strat Binding donor well positive SEQ
ID position sequence Score number number CTL line NO 322 LYEKaNTPEL
330 0/1 0/8 -- 98 435 NYVNiLATII 90 0/1 0/8 -- 99 40 SYVEeEMPQI 90
1/1 100 101 DFQDsVFNDL 72.576 1/1 101 866 SFFErRSHTL 24 0/1 0/8 --
102 811 SFSKtPKRAL 20 0/1 1/8 0/1 103 268 KYLPlGDERC 18 0/1 0/8 --
104 84 EFEGlDSPEF 16.5 0/1 1/8 0/1 105 236 KVPPfQDCIL 14.4 0/1 0/8
-- 106 728 RPPTeQANVL 14.4 0/1 0/8 -- 107 507 KYSKdLVKTY 12 0/1 0/8
-- 108 281 VVEEnIVKDL 10.08 0/1 0/8 -- 109
TABLE-US-00009 TABLE 5A HLA-A*2402 binding 9-mer peptides derived
from HIG2 positive positive strat Binding donor well positive SEQ
ID position sequence Score number number CTL line NO 19 IFVRVMESL
42 1/3 110 22 RVMESLEGL 14.4 1/3 111 8 YLLGVVLTL 8.4 1/3 387 7
LYLLGVVLT 7.5 0/2 3/15 0/3 388 23 VMESLEGLL 7.2 0/2 0/16 -- 389 9
LLGVVLTLL 5.6 -- -- -- 390
TABLE-US-00010 TABLE 5B Table 5B HLA-A*2402 binding 10-mer peptides
derived from HIG2 posi- posi- strat tive tive SEQ posi- Binding
donor well positive ID tion sequence Score number number CTL line
NO 7 LYLLGVVLTL 420 1/3 112 22 RVMESLEGLL 17.28 0/3 4/24 0/4 113 8
YLLGVVLTLL 8.4 -- -- -- 391 5 LNLYLLGVVL 7.2 0/2 0/12 -- 392 46
LANTEPTKGL 6 0/2 0/14 -- 393 18 SIFVRVMESL 5.6 1/2 394
TABLE-US-00011 TABLE 5C HLA-A*0201 binding 9-mer peptides derived
from HIG2 posi- strat positive tive SEQ posi- Binding donor well
positive ID tion sequence Score number number CTL line NO 8
YLLGVVLTL 836.253 1/1 114 13 VLTLLSIFV 650.311 0/1 0/12 -- 115 15
TLLSIFVRV 488.951 1/1 116 4 VLNLYLLGV 271.948 1/1 117 9 LLGVVLTLL
83.527 0/1 0/12 -- 118 22 RVMESLEGL 31.957 0/1 0/12 -- 119 6
NLYLLGVVL 28.027 0/1 0/12 -- 120
TABLE-US-00012 TABLE 5D HLA-A*0201 binding 10-mer peptides derived
from HIG2 posi- posi- strat tive tive SEQ posi- Binding donor well
positive ID tion sequence Score number number CTL line NO 8
YLLGvVLTLL 836.253 1/1 121 12 VVLTlLSIFV 210.538 -- -- -- 122 29
GLLEsPSPGT 113.047 0/1 0/12 -- 123 6 NLYLlGVVLT 54.847 -- -- -- 124
4 VLNLyLLGVV 14.495 0/1 0/12 -- 125 15 TLLSiFVRVM 13.174 0/1 0/12
-- 126 18 SIFVrVMESL 12.248 0/1 0/12 -- 127 14 LTLLsIFVRV 11.545 --
-- -- 128
TABLE-US-00013 TABLE 6A HLA-A*2402 binding 9-mer peptides derived
from INHBB posi- posi- tive tive SEQ strat Binding donor well
positive ID position sequence Score number number CTL line NO 383
LYFDDEYNI 60 0/3 0/20 -- 129 238 LFERGERRL 30 0/3 1/19 0/1 130 7
RALGAACLL 12 0/3 0/21 -- 131 388 EYNIVKRDV 10.5 0/3 0/18 -- 132 180
LYLKLLPYV 9 1/2 395 163 ISNEGNQNL 8.64 0/1 0/8 -- 396 223 RSGWHTFPL
8 0/1 0/6 -- 397 176 ASLWLYLKL 7.92 0/1 0/7 -- 398 338 AYLAGVPGS
7.5 0/1 1/7 0/1 399 213 NMVEKRVDL 7.2 0/1 0/8 -- 400 102 AMVTALRKL
6.6 0/1 0/8 -- 401 250 VQCDSCQEL 6.336 0/1 0/8 -- 402 369 NSCCIPTKL
6.16 0/1 0/8 -- 403 330 NYCEGSCPA 6 0/1 0/7 -- 404 172 FVVQASLWL 6
0/1 0/8 -- 405 355 VNQYRMRGL 6 0/1 0/8 -- 406 307 QFFIDFRLI 6 0/1
0/7 -- 407 14 LLLLAAGWL 6 -- -- -- 408 306 QQFFIDFRL 5.6 0/1 0/6 --
409 170 NLFVVQASL 5.6 0/1 0/7 -- 410 327 YYGNYCEGS 5 0/1 1/8 0/1
411
TABLE-US-00014 TABLE 6B HLA-A*2402 binding 10-mer peptides derived
from INHBB posi- posi- strat tive tive SEQ posi- Binding donor well
positive ID tion sequence Score number number CTL line NO 180
LYLKLLPYVL 360 1/3 133 171 LFVVQASLWL 30 -- -- -- 134 305
RQQFFIDFRL 16.8 1/3 135 73 DFLEAVKRHI 12.6 0/3 4/20 0/4 136 7
RALGAACLLL 12 1/3 137 273 RPFVVVQARL 11.2 0/3 1/20 0/1 138 338
AYLAGVPGSA 10 0/3 2/20 0/2 139 169 QNLFvVQASL 8.4 0/1 1/6 0/1 412
249 DVQCdSCQEL 7.92 0/1 4/6 0/4 413 383 LYFDdEYNIV 7.2 0/1 0/6 --
415 229 FPLTeAIQAL 7.2 0/1 1/6 0/1 416 299 RTNLcCRQQF 7.2 0/1 5/6
0/5 417 101 AAMVtALRKL 6.6 0/1 2/6 0/2 418 368 VNSCcIPTKL 6.16 0/1
2/6 0/2 419 13 CLLLlAAGWL 6 -- -- -- 420 354 VVNQyRMRGL 6 0/1 0/6
-- 421 150 DGLAsSRVRL 6 0/1 2/6 0/2 422 293 GLECdGRTNL 6 0/1 0/6
423 330 NYCEgSCPAY 6 0/1 1/6 0/1 424 176 ASLWlYLKLL 6 0/1 1/6 0/1
425 212 WNMVeKRVDL 6 1/1 426 74 FLEAvKRHIL 6 0/1 2/6 0/2 427 331
YCEGsCPAYL 6 0/1 1/6 0/1 428 77 AVKRhILSRL 5.6 0/1 1/6 0/1 429 175
QASLwLYLKL 5.28 0/1 2/6 0/2 430 326 GYYGnYCEGS 5 0/1 1/6 0/1 431
159 LYFFiSNEGN 5 0/1 4/6 0/4 432 327 YYGNyCEGSC 5 0/1 1/6 0/1
433
TABLE-US-00015 TABLE 6C HLA-A*0201 binding 9-mer peptides derived
from INHBB posi- posi- tive tive SEQ strat Binding donor well
positive ID position sequence Score number number CTL line NO 177
SLWLYLKLL 407.808 0/1 0/8 140 14 LLLLAAGWL 96.074 -- -- -- 141 170
NLFVVQASL 79.041 0/1 0/8 142 213 NMVEKRVDL 63.256 0/1 0/8 143 172
FVVQASLWL 47.291 0/1 0/8 144 306 QQFFIDFRL 46.48 0/1 0/8 145 281
RLGDSRHRI 42.774 0/1 0/8 146 174 VQASLWLYL 34.427 0/1 0/8 147 257
ELAVVPVFV 28.69 0/1 1/8 0/1 148 313 RLIGWNDWI 28.116 0/1 1/8 0/1
149 139 RVSEIISFA 22.546 0/1 3/8 0/3 150 151 GLASSRVRL 21.362 0/1
0/8 151 8 ALGAACLLL 21.362 0/1 1/8 0/1 152 250 VQCDSCQEL 15.096 0/1
1/8 0/1 153
TABLE-US-00016 TABLE 6D Table 6D HLA-A*0201 binding 10-mer peptides
derived from INHBB posi- posi- strat tive tive SEQ posi- Binding
donor well positive ID tion sequence Score number number CTL line
NO 179 WLYLKLLPYV 12951.1 0/1 1/8 0/1 154 301 NLCCRQQFFI 332.806
0/1 0/8 155 237 ALFERGERRL 64.814 0/1 0/8 156 382 MLYFDDEYNI 56.754
0/1 0/8 157 13 CLLLLAAGWL 56.514 -- -- -- 158 8 ALGAACLLLL 49.134
-- -- -- 159 313 RLIGWNDWII 32.081 0/1 0/8 160 173 VVQASLWLYL
29.711 0/1 2/8 0/2 161 256 QELAVVPVFV 27.521 0/1 0/8 162 162
FISNEGNQNL 13.512 0/1 1/8 0/1 163 305 RQQFFIDFRL 12.562 0/1 0/8 164
362 GLNPGTVNSC 11.426 0/1 0/7 165 85 RLQMRGRPNI 10.433 0/1 1/8 0/1
166 69 RVDGDFLEAV 10.425 0/1 0/8 167
TABLE-US-00017 TABLE 7A HLA-A*2402 binding 9-mer peptides derived
from KIF20A posi- posi- tive tive SEQ strat Binding donor well
positive ID position sequence Score number number CTL line NO 308
IYNELLYDL 432 0/2 0/14 -- 168 621 MYEEKLNIL 432 0/2 0/14 -- 169 67
VYLRVRPLL 420 0/2 0/14 -- 170 499 KFSAIASQL 56 0/2 0/14 -- 171 304
SFFEIYNEL 44.352 0/2 0/14 -- 172 187 IFNSLQGQL 36 0/2 0/14 -- 173
305 FFEIYNELL 30 1/2 174 23 MFESTAADL 30 0/2 0/14 -- 175 256
SFDSGIAGL 20 0/2 0/14 -- 176 298 RFSIWISFF 20 -- -- -- 177 383
IFSIRILHL 20 1/2 178 647 KIEELEALL 17.28 0/2 0/14 -- 179 625
KLNILKESL 14.4 0/2 0/14 -- 180 695 KLQQCKAEL 13.2 0/2 0/14 -- 181
726 FTIDVDKKL 11.088 0/2 0/14 -- 182 688 QLQEVKAKL 11.088 0/2 0/14
-- 183
TABLE-US-00018 TABLE 7B HLA-A*2402 binding 10-mer peptides derived
from KIF20A posi- posi- strat tive tive SEQ posi- Binding donor
well positive ID tion sequence Score number number CTL line NO 308
IYNElLYDLL 432 0/2 0/14 -- 184 182 RSLAlIFNSL 24.192 0/2 1/14 0/1
185 304 SFFEiYNELL 24 1/2 186 742 RLLRtELQKL 15.84 0/2 0/14 -- 187
739 KNIRlLRTEL 15.84 0/2 0/14 -- 188 218 RQEEmKKLSL 14.4 0/2 2/14
0/2 189 70 RVRPlLPSEL 12.672 0/2 0/14 -- 190 871 RILRsRRSPL 12 0/2
0/14 -- 191 89 RIENvETLVL 12 0/2 1/14 0/1 192 364 KNQSfASTHL 12 0/2
0/14 -- 193 66 KVYLrVRPLL 11.2 1/2 194 60 DSMEkVKVYL 10.08 0/2 0/14
-- 195
TABLE-US-00019 TABLE 8A HLA-A*2402 binding 9-mer peptides derived
from KNTC2 posi- posi- tive tive SEQ strat Binding donor well
positive ID position sequence Score number number CTL line NO 309
KYQAYMSNL 600 1/3 196 457 VYVPLKELL 432 0/3 0/18 -- 197 414
EYHKLARKL 264 0/3 0/18 -- 198 139 SYELPDTKF 165 0/3 0/18 -- 199 629
KYEKKATLI 150 0/3 0/18 -- 200 400 KYARGKEAI 100 0/3 1/18 0/1 201
124 DFLKIFTFL 50.4 1/3 202 134 GFLCPSYEL 33 0/3 0/18 -- 203 257
LFNVDAFKL 33 0/3 0/18 -- 204 242 SFDEMNAEL 26.4 0/3 0/18 -- 205 128
IFTFLYGFL 24 0/3 0/18 -- 206 146 KFEEEVPRI 18 0/3 1/18 0/1 207 368
RINHERNEL 15.84 0/3 1/18 0/1 208 235 SFMSGADSF 15 0/3 0/18 -- 209
154 IFKDLGYPF 14.4 1/3 210 563 EYQLVVQTT 12.6 0/3 0/18 -- 211 474
KALNKKMGL 12 0/3 1/18 0/1 212 150 EVPRIFKDL 10.08 1/3 213
TABLE-US-00020 TABLE 8B HLA-A*2402 binding 10-mer peptides derived
from KNTC2 posi- posi- strat tive tive SEQ posi- Binding donor well
positive ID tion sequence Score number number CTL line NO 452
KYRAQVYVPL 560 2/3 214 610 EYEECMSEDL 360 0/3 1/18 0/1 215 360
KYSVADIERI 100 0/3 0/18 -- 216 227 DYTIKCYESF 100 1/3 217 146
KFEEEVPRIF 50.4 0/3 0/18 -- 218 90 AFIQQCIRQL 30 0/3 0/18 -- 219 20
RSQDVNKQGL 17.28 0/3 1/18 0/1 220 501 RTLKEEVQKL 15.84 0/3 0/18 --
221 403 RGKEAIETQL 13.44 0/3 1/18 0/1 222 273 RALNEQIARL 12 1/3 223
563 EYQLVVQTTT 10.5 0/3 3/22 0/3 224 467 ETEEEINKAL 10.08 0/3 1/22
0/1 225 541 LLESTVNQGL 10.08 0/3 1/22 0/1 226
TABLE-US-00021 TABLE 9A HLA-A*0201 binding 9-mer peptides derived
from TTK posi- posi- tive tive SEQ strat Binding donor well
positive ID position sequence Score number number CTL line NO 462
YMSCFRTPV 878.055 1/1 227 547 KQIYAIKYV 312.218 1/1 228 630
NMLEAVHTI 262.897 0/1 1/8 0/1 229 278 LLNSPDCDV 118.238 0/1 1/8 0/1
230 498 ILATPLQNL 83.527 0/1 0/8 -- 231 811 YVLGQLVGL 73.172 0/1
0/8 -- 232 719 SLGCILYYM 62.845 1/2 233 670 QMQPDTTSV 50.232 0/1
0/8 -- 234 804 GTTEEMKYV 50.102 0/1 0/8 -- 235 654 LIVDGMLKL 47.088
0/1 1/8 0/1 236 363 SLLAKLEET 31.074 0/1 0/8 -- 237 790 YVQIQTHPV
27.995 0/1 0/8 -- 238 785 LLAHPYVQI 26.604 0/1 0/8 -- 239 86
KLIGRYSQA 26.082 0/1 0/8 -- 240 186 NLNLQKKQL 21.362 0/1 0/8 -- 241
671 MQPDTTSVV 20.152 0/1 0/8 -- 242 577 KLQQHSDKI 17.892 0/1 0/8 --
243 142 FAFVHISFA 14.856 0/1 0/8 -- 244 322 CELRNLKSV 11.509 0/1
0/8 -- 245 824 SILKAAKTL 10.868 0/1 0/8 -- 246
TABLE-US-00022 TABLE 9B HLA-A*0201 binding 10-mer peptides derived
from TTK posi- posi- strat tive tive SEQ posi- Binding donor well
positive ID tion sequence Score number number CTL line NO 68
LLLKLEKNSV 437.482 0/1 0/8 -- 247 277 NLLNSPDCDV 257.342 0/1 0/8 --
248 653 FLIVDGMLKL 226.014 0/1 0/8 -- 249 423 TTFEQPVFSV 195.487
0/1 0/8 -- 250 542 VLNEKKQIYA 190.448 0/1 0/8 -- 251 658 GMLKLIDFGI
161.697 0/1 0/8 -- 252 194 LLSEEEKKNL 148.896 0/1 0/8 -- 253 462
YMSCFRTPVV 94.738 1/1 254 57 MMANNPEDWL 70.685 0/1 0/8 -- 255 600
MVMECGNIDL 48.205 0/1 0/8 -- 256 689 YMPPEAIKDM 37.961 0/1 0/8 --
257 86 KLIGRYSQAI 36.515 0/1 0/8 -- 258 669 NQMQPDTTSV 26.092 0/1
1/8 0/1 259 497 QILATPLQNL 24.997 0/1 0/8 -- 260 654 LIVDGMLKLI
22.997 0/1 0/8 -- 261 186 NLNLQKKQLL 21.362 0/1 1/8 0/1 262 670
QMQPDTTSVV 20.595 0/1 0/8 -- 263 803 KGTTEEMKYV 20.102 0/1 0/8 --
264 11 LTIDSIMNKV 15.486 0/1 0/8 -- 265 577 KLQQHSDKII 14.971 0/1
0/8 -- 266
TABLE-US-00023 TABLE 10A HLA-A*0201 binding 9-mer peptides derived
from URLC10 posi- posi- tive tive SEQ strat Binding donor well
positive ID position sequence Score number number CTL line NO 131
KIFPRFFMV 1364.78 0/1 0/8 -- 267 204 GLWLAILLL 407.808 0/1 0/8 --
268 65 LLVVALPRV 271.948 0/1 0/8 -- 269 60 ALLALLLVV 242.674 -- --
-- 270 206 WLAILLLLA 52.561 1/1 271 212 LLASIAAGL 36.316 1/1 272
210 LLLLASIAA 31.249 0/1 0/8 -- 273 137 FMVAKQCSA 16.505 0/1 2/8
0/2 274 58 TMALLALLL 15.428 0/1 2/8 0/2 275 59 MALLALLLV 13.975 0/1
2/8 0/2 276 209 ILLLLASIA 12.812 0/1 0/8 -- 434 208 AILLLLASI
12.208 -- -- -- 277 69 ALPRVWTDA 8.446 0/1 0/8 -- 278 197 SMGESCGGL
8.223 0/1 0/8 -- 279 61 LLALLLVVA 7.964 -- -- -- 280 67 VVALPRVWT
6.097 0/1 0/8 -- 281 72 RVWTDANLT 5.412 0/1 0/8 -- 282 160
FLLEEPMPF 5.2 0/1 1/8 0/1 283 62 LALLLVVAL 4.292 0/1 0/8 -- 284 57
GTMALLALL 2.525 0/1 1/8 0/1 285
TABLE-US-00024 TABLE 10B HLA-A*0201 binding 10-mer peptides derived
from URLC10 posi- posi- strat tive tive SEQ posi- Binding donor
well positive ID tion sequence Score number number CTL line NO 64
LLLVVALPRV 1006.21 0/1 0/8 -- 286 204 GLWLAILLLL 407.808 0/1 1/8
0/1 287 211 LLLASIAAGL 134.369 1/1 288 258 TMALLALLLV 115.534 -- --
-- 289 61 LLALLLVVAL 83.527 -- -- -- 290 160 FLLEEPMPFF 65.782 0/1
0/8 -- 291 209 ILLLLASIAA 31.249 0/1 0/8 -- 292 131 KIFPRFFMVA
26.186 0/1 0/8 -- 293 60 ALLALLLVVA 17.334 -- -- -- 294 66
LVVALPRVWT 6.097 0/1 0/8 -- 295 59 MALLALLLVV 5.73 -- -- -- 296 2
RLQRPRQAPA 4.968 0/1 1/8 0/1 297 112 CQNPRRCKWT 4.156 0/1 0/8 --
298 72 RVWTDANLTA 3.608 0/1 0/8 -- 299 53 WAPLGTMALL 3.139 0/1 0/8
-- 300 121 TEPYCVIAAV 3.111 0/1 0/8 -- 301 162 LEEPMPFFYL 2.739 0/1
1/8 0/1 302 181 LEGPPINSSV 2.299 0/1 2/8 0/2 303 170 YLKCCKIRYC
2.024 0/1 0/8 -- 304 130 VKIFPRFFMV 1.81 0/1 0/8 -- 305
Stimulation of the T Cells Using the Predicted Peptides from CDH3
Restricted with HLA-A*2402 and Establishment for CTL Lines
Stimulated with CDH3 Derived Peptides
[0455] CTLs for those peptides derived from CDH3 were generated
according to the protocols set forth in "Materials and Methods"
section above. Resulting that CTLs having detectable specific CTL
activity, as determined by IFN-gamma ELISPOT assay, are shown in
FIG. 1. In particular, CDH3-A24-9-513 (SEQ ID NO: 19),
CDH3-A24-9-406 (SEQ ID NO: 22), CDH3-A24-10-807 (SEQ ID NO: 30),
CDH3-A24-10-332 (SEQ ID NO: 34), CDH3-A24-10-655 (SEQ ID NO: 344)
and CDH3-A24-10-470 (SEQ ID NO: 358) demonstrated potent IFN-gamma
production as compared to the control by IFN-gamma ELISPOT assay,
and the cells in the positive well number #5 stimulated with SEQ ID
NO: 19, #2 with SEQ ID NO: 22, #5 with SEQ ID NO: 30, #4 with SEQ
ID NO: 34, #1 with SEQ ID NO: 344 and #4 with SEQ ID NO: 358 were
expanded and CTL lines were established. Those CTL lines having
higher specific CTL activities against the peptide-pulsed target as
compared to the activities against target without peptide pulse
were determined by ELISA. Results are shown in FIG. 1. While, other
peptides shown in table 2 could not establish the CTL lines despite
possible binding activity with HLA-A*2402. For example, the typical
negative peptide (CDH3-A24-10-248) were shown in FIG. 1a. In this
invention, the peptides which could establish CTL line were
selected as potent CTL stimulation peptide.
Establishment for CTL Clones Stimulated with CDH3 Derived
Peptides
[0456] Furthermore, the limiting dilution from these CTL lines was
performed according to the protocols set forth in the "Materials
and Methods" section above. The establishment of CTL clones from
CDH3-A24-10-807 (SEQ ID NO: 30) #5 and CDH3-A24-10-655 (SEQ ID NO:
344) #1 CTL line are shown in FIG. 1f and g. CTL clones had potent
and specific CTL activities against the peptide-pulsed target as
compared to the activities against target without peptide
pulse.
Specific CTL Activity Against the Target Cells Expressing CDH3 and
HLA-A*2402
[0457] The established CTL line raised against these peptides were
examined for their ability to recognize the target cells expressing
CDH3 and HLA-A*2402. Specific CTL activity against COS7 transfected
with both full length CDH3 gene and the HLA-A*2402 molecule, which
serves as a specific model for the target cells endogenously
express CDH3 and HLA-A*2402, was tested using as effector cells the
CTL lines raised by CDH3-A24-10-807 (SEQ ID NO: 30) and
CDH3-A24-10-655 (SEQ ID NO: 344). COS7 transfected with full length
CDH3 but not HLA-A*2402 and COS7 transfected with HLA-A*2402 but
not full length CDH3 were prepared as controls. The CTL clones
demonstrated the highest specific CTL activity against COS7 that
was transfected with both CDH3 and HLA-A2402 (Figure if and g).
[0458] These results clearly demonstrate that CDH3-A24-10-807 (SEQ
ID NO: 30) and CDH3-A24-10-655 (SEQ ID NO: 344) are naturally
expressed on the target cell surface with HLA-A2402 molecule and
recognize CTL. Furthermore, these peptides are epitope peptides,
which may serve as cancer vaccines targeting CDH3 expressed
tumors.
Stimulation of the T Cells Using the Predicted Peptides from EPHA4
Restricted with HLA-A*2402 or HLA-A*0201, and Establishment for CTL
Lines Stimulated with EPHA4 Derived Peptides
[0459] CTLs for those peptides derived from EphA4 were generated by
IFN-gamma ELISPOT assay. Resulting that CTLs having detectable
specific CTL activity, as determined by IFN-gamma ELISPOT assay,
are shown in FIG. 2. In particular, EphA4-A24-9-453 (SEQ ID NO:
41), EphA4-A24-9-5 (SEQ ID NO: 44), EphA4-A24-9-869 (SEQ ID NO:
46), EphA4-A24-9-420 (SEQ ID NO: 48), EphA4-A24-10-24 (SEQ ID NO:
78), EphA4-A02-9-501 (SEQ ID NO: 376) and EphA4-A02-9-165 (SEQ ID
NO: 379) demonstrated potent IFN-gamma production by IFN-gamma
ELISPOT assay, and the cells in the positive well number #3
stimulated with EphA4-A24-9-453 (SEQ ID NO: 41), #2 with
EphA4-A24-9-5 (SEQ ID NO: 44), #5 with EphA4-A24-9-869 (SEQ ID NO:
46), #6 with EphA4-A24-9-420 (SEQ ID NO: 48), #4 with
EphA4-A24-10-24 (SEQ ID NO: 78), #8 with EphA4-A02-9-501 (SEQ ID
NO: 376) and #3 with EphA4-A02-9-165 (SEQ ID NO: 379) were expanded
and CTL lines were established. Those CTL lines having higher
specific CTL activities against the peptide-pulsed target as
compared to the activities against target without peptide pulse
were determined by ELISA. Especially, CTL lines stimulated with
EphA4-A02-9-501 (SEQ ID NO: 376) and EphA4-A02-9-165 (SEQ ID NO:
379) were tested by 51Cr-release assay according to the protocols
set forth in the "Materials and Methods" section above. Results are
shown in FIG. 2a-h. While, other peptides shown in table 3 could
not establish the CTL lines despite possible binding activity with
HLA-A*2402 or HLA-A*0201. For example, the typical negative peptide
(EphA4-A24-9-384) were shown in FIG. 2a. In this invention, the
peptides which could establish CTL line were selected as potent CTL
stimulation peptides.
Stimulation of the T Cells Using the Predicted Peptides from ECT2
Restricted with HLA-A*2402, and Establishment for CTL Lines
Stimulated with ECT2 Derived Peptides
[0460] CTLs for those peptides derived from ECT2 were generated
according to the protocols set forth in the "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity as determined by an IFN-gamma ELISPOT assay are shown in
FIG. 3. In particular, ECT2-A24-9-515 (SEQ ID NO: 80),
ECT2-A24-10-40 (SEQ ID NO: 100) and ECT2-A24-10-101 (SEQ ID NO:
101) showed potent IFN-gamma production, and the cells in the
positive well number #7 stimulated with ECT2-A24-9-515 (SEQ ID NO:
80), #2 with ECT2-A24-10-40 (SEQ ID NO: 100) and #1 with
ECT2-A24-10-101 (SEQ ID NO: 101) were expanded and CTL lines were
established. Those CTL lines having higher specific CTL activities
against the peptide-pulsed target as compared to the activities
against target without peptide pulse were determined by ELISA.
Results are shown in FIG. 3a-d. While, other peptides shown in
table 4 could not establish the CTL lines despite possible binding
activity with HLA-A*2402. For example, the typical negative peptide
(ECT2-A24-10-322, ECT2-A24-9-657 and ECT2-A24-10-811) were shown in
FIG. 2a. In this invention, the peptides which could establish CTL
line were selected as potent CTL stimulation peptide.
Establishment for CTL Clones Stimulated with ECT2 Derived
Peptides
[0461] Furthermore, the limiting dilution from these CTL lines was
performed according to the protocols set forth in the "Materials
and Methods" section above. The establishment of CTL clones from
ECT2-A24-10-40 (SEQ ID NO: 100) #2 CTL line are shown in FIG. 3c.
CTL clones had potent and specific CTL activities against the
peptide-pulsed target as compared to the activities against target
without peptide pulse.
Specific CTL Activity Against the Target Cells Expressing ECT2 and
HLA-A*2402
[0462] The established CTL line raised against these peptides were
examined for their ability to recognize the target cells expressing
ECT2 and HLA-A*2402. Specific CTL activity against COS7 transfected
with both full length ECT2 gene and the HLA-A*2402 molecule, which
serves as a specific model for the target cells endogenously
express ECT2 and HLA-A*2402, was tested using as effector cells the
CTL clone raised by ECT2-A24-10-40 (SEQ ID NO: 100) and the CTL
line raised by ECT2-A24-10-101 (SEQ ID NO: 101). COS7 transfected
with full length ECT2 but not HLA-A*2402 and COS7 transfected with
HLA-A*2402 but not full length ECT2 (replaced other gene e.g.
URLC10 or INHBB) were prepared as controls. The CTL line
demonstrating the highest specific CTL activity against COS7 that
was transfected with both ECT2 and HLA-A2402 (FIGS. 3c and d).
[0463] These results clearly demonstrate that ECT2-A24-10-40 (SEQ
ID NO: 100) and ECT2-A24-10-101 (SEQ ID NO: 101) are naturally
expressed on the target cell surface with HLA-A2402 molecule and
recognize CTL. Furthermore, these peptides are epitope peptides,
which may serve as cancer vaccines targeting ECT2 expressed
tumors.
Cytotoxic Activity Against Cancer Cell Line Endogenously Expressing
HLA-A*2402 and ECT2
[0464] Furthermore, Cytotoxic activity was examined by cytotoxicity
assay according to the protocols set forth in the "Materials and
Methods" section above. As a result, as shown in FIG. 3b, CTL clone
stimulated with ECT2-A24-9-515 (SEQ ID NO: 80) showed remarkably
high cytotoxic effect towards HLA-A24-positive and ECT-positive
cancer cell lines TE6, compared to that towards HLA-A24-negative
and ECT-positive cancer cell lines TE5.
Stimulation of the T Cells Using the Predicted Peptides from HIG2
Restricted with HLA-A*2402 or HLA-A*0201, and Establishment for CTL
Lines Stimulated with HIG2 Derived Peptides
[0465] CTLs for those peptides derived from HIG2 were generated
according to the protocols set forth in the "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity as determined by an IFN-gamma ELISPOT assay are shown in
FIG. 4. In particular, HIG2-A24-9-19 (SEQ ID NO: 110),
HIG2-A24-9-22 (SEQ ID NO: 111), HIG2-A24-9-8 (SEQ ID NO: 387),
HIG2-A24-10-7 (SEQ ID NO: 112), HIG2-A24-10-18 (SEQ ID NO: 394),
HIG2-A02-9-8 (SEQ ID NO: 114), HIG2-A02-9-15 (SEQ ID NO: 116),
HIG2-A02-9-4 (SEQ ID NO: 117) and HIG2-A02-10-8 (SEQ ID NO: 121)
demonstrated potent IFN-gamma production by IFN-gamma ELISPOT
assay, and the cells in the positive well number #6 stimulated with
HIG2-A24-9-19 (SEQ ID NO: 110), #7 with HIG2-A24-9-22 (SEQ ID NO:
111), #5 with HIG2-A24-9-8 (SEQ ID NO: 387), #1 with HIG2-A24-10-7
(SEQ ID NO: 112), #7 with HIG2-A24-10-18 (SEQ ID NO: 394), #10 with
HIG2-A02-9-8 (SEQ ID NO: 114), #10 with HIG2-A02-9-15 (SEQ ID NO:
116), #10 with HIG2-A02-9-4 (SEQ ID NO: 117) and #9 with
HIG2-A02-10-8 (SEQ ID NO: 121) were expanded and CTL lines were
established. Those CTL lines having higher specific CTL activities
against the peptide-pulsed target as compared to the activities
against target without peptide pulse were determined by ELISA.
Results are shown in FIG. 4a-j. While, other peptides shown in
table 5 could not establish the CTL lines despite possible binding
activity with HLA-A*2402. For example, the typical negative peptide
(HIG2-A24-9-7) were shown in FIG. 4a. In this invention, the
peptides which could establish CTL line were selected as potent CTL
stimulation peptide.
Establishment for CTL Clones Stimulated with HIG2 Derived
Peptides
[0466] Furthermore, the limiting dilution from these CTL lines was
performed according to the protocols set forth in the "Materials
and Methods" section above. The establishment of CTL clones from
HIG2-A24-9-22 (SEQ ID NO: 111) #7 CTL line, HIG2-A24-9-8 (SEQ ID
NO: 387) #5 CTL line, HIG2-A24-10-7 (SEQ ID NO: 112) #1 CTL line,
HIG2-A24-10-18 (SEQ ID NO: 394) #7 CTL line and HIG2-A02-9-4 (SEQ
ID NO: 117) #10 CTL line are shown in FIGS. 4c, e, f, g and i. CTL
clones had potent and specific CTL activities against the
peptide-pulsed target as compared to the activities against target
without peptide pulse.
Specific CTL Activity Against the Target Cells Expressing HIG2 and
HLA-A*0201
[0467] The established CTL line raised against these peptides were
examined for their ability to recognize the target cells expressing
HIG2 and HLA-A*0201. Specific CTL activity against 293T or COS7
transfected with both full length HIG2 gene and the HLA-A*0201
molecule, which serves as a specific model for the target cells
endogenously express HIG2 and HLA-A*0201, was tested using as
effector cells the CTL lines raised by HIG2-A02-9-8 (SEQ ID NO:
114), HIG2-A02-9-15 (SEQ ID NO: 116) and the CTL clone raised by
HIG2-A02-9-4 (SEQ ID NO: 117). 293T or COS7 transfected with full
length ECT2 but not HLA-A*0201 and 293T or COS7 transfected with
HLA-A*0201 but not full length ECT2 (or replaced other gene e.g.
FoxP3 or TTK) were prepared as controls. The CTL line demonstrating
the highest specific CTL activity against 293T or COS7 that was
transfected with both ECT2 and HLA-A*0201 (FIGS. 4e, h and i).
[0468] These results clearly demonstrate that HIG2-A02-9-8 (SEQ ID
NO: 114), HIG2-A02-9-15 (SEQ ID NO: 116) and HIG2-A02-9-4 (SEQ ID
NO: 117) are naturally expressed on the target cell surface with
HLA-A2402 or HLA-A0201 molecule and recognize CTL. Furthermore,
these peptides are epitope peptides, which may serve as cancer
vaccines targeting HIG2 expressed tumors.
Cytotoxic Activity Against Cancer Cell Line Endogenously Expressing
HLA-A*0201 and HIG2
[0469] Furthermore, Cytotoxic activity was examined by cytotoxicity
assay according to the protocols set forth in the "Materials and
Methods" section above. As a result, as shown in FIG. 4i, CTL clone
stimulated with HIG2-A02-9-4 (SEQ ID NO: 117) showed remarkably
high cytotoxic effect towards HLA-A02-positive and HIG2-positive
cancer cell lines CAki-1, compared to that towards HLA-A02-negative
and HIG2-positive cancer cell lines A498.
Stimulation of the T Cells Using the Predicted Peptides from INHBB
Restricted with HLA-A*2402 or HLA-A*0201, and Establishment for CTL
Lines Stimulated with INHBB Derived Peptides
[0470] CTLs for those peptides derived from INHBB were generated
according to the protocols set forth in the "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity as determined by an IFN-gamma ELISPOT assay are shown in
FIG. 5. In particular, INHBB-A24-9-180 (SEQ ID NO: 395),
INHBB-A24-10-180 (SEQ ID NO: 133), INHBB-A24-10-305 (SEQ ID NO:
135), INHBB-A24-10-7 (SEQ ID NO: 137) and INHBB-A24-10-212 (SEQ ID
NO: 426) demonstrated potent IFN-gamma production by IFN-gamma
ELISPOT assay, and the cells in the positive well number #7
stimulated with INHBB-A24-9-180 (SEQ ID NO: 395), #3 with
INHBB-A24-10-180 (SEQ ID NO: 133), #2 with INHBB-A24-10-305 (SEQ ID
NO: 135), #8 and #2 with INHBB-A24-10-7 (SEQ ID NO: 137) and #1
with INHBB-A24-10-212 (SEQ ID NO: 426) were expanded and CTL lines
were established. Those CTL lines having higher specific CTL
activities against the peptide-pulsed target as compared to the
activities against target without peptide pulse were determined by
ELISA. Results are shown in FIG. 5b-e. While, other peptides shown
in table 6 could not establish the CTL lines despite possible
binding activity with HLA-A*2402 and HLA*0201. For example, the
typical negative peptide (INHBB-A24-9-238) were shown in FIG. 5a.
In this invention, the peptides which could establish CTL line were
selected as potent CTL stimulation peptide.
Establishment for CTL Clones Stimulated with INHBB Derived
Peptides
[0471] Furthermore, the limiting dilution from these CTL lines was
performed according to the protocols set forth in the "Materials
and Methods" section above. The establishment of CTL clones from
INHBB-A24-9-180 (SEQ ID NO: 395) #7 CTL line, and INHBB-A24-10-305
(SEQ ID NO: 135) #2 CTL line are shown in FIGS. 5b and d. CTL
clones had potent and specific CTL activities against the
peptide-pulsed target as compared to the activities against target
without peptide pulse.
Specific CTL Activity Against the Target Cells Expressing INHBB and
HLA-A*2402
[0472] The established CTL line raised against these peptides were
examined for their ability to recognize the target cells expressing
INHBB and HLA-A*2402. Specific CTL activity against 293T
transfected with both full length INHBB gene and the HLA-A*2402
molecule, which serves as a specific model for the target cells
endogenously express INHBB and HLA-A*2402, was tested using as
effector cells the CTL lines raised by INHBB-A24-10-180 (SEQ ID NO:
133) and INHBB-A24-10-7 (SEQ ID NO: 137) and the CTL clone raised
by INHBB-A24-10-305 (SEQ ID NO: 135). 293T transfected with full
length INHBB but not HLA-A*2402 and 293T transfected with
HLA-A*2402 but not full length INHBB were prepared as controls. The
CTL line demonstrating the highest specific CTL activity against
293T was that transfected with both INHBB and HLA-A*2402 (FIGS. 5c,
d and e).
[0473] These results clearly demonstrate that INHBB-A24-10-305 (SEQ
ID NO: 135), INHBB-A24-10-180 (SEQ ID NO: 133) and INHBB-A24-10-7
(SEQ ID NO: 137) are naturally expressed on the target cell surface
with HLA-A2402 molecule and recognize CTL.
[0474] Furthermore, these peptides are epitope peptides, which may
serve as cancer vaccines targeting INHBB expressed tumors.
Cytotoxic Activity Against Cancer Cell Line Endogenously Expressing
HLA-A*2402 and INHBB
[0475] Furthermore, Cytotoxic activity was performed by
cytotoxicity assay according to the protocols set forth in the
"Materials and Methods" section above. As a result, as shown in
FIG. 5b, CTL clone stimulated with INHBB-A24-9-180 (SEQ ID NO: 395)
showed remarkably high cytotoxic effect towards HLA-A24-positive
and INHBB-positive cancer cell lines MIAPaca2, compared to that
towards HLA-A24-negative and INHBB-positive cancer cell lines
CAki-2.
Stimulation of the T Cells Using the Predicted Peptides from KIF20A
Restricted with HLA-A*2402, and Establishment for CTL Lines
Stimulated with KIF20A Derived Peptides
[0476] CTLs for those peptides derived from KIF20A were generated
according to the protocols set forth in the "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity as determined by an IFN-gamma ELISPOT assay are shown in
FIG. 6. In particular, KIF20A-A24-9-305 (SEQ ID NO: 174),
KIF20A-A24-9-383 (SEQ ID NO: 178), KIF20A-A24-10-304 (SEQ ID NO:
186) and KIF20A-A24-10-66 (SEQ ID NO: 194) demonstrated potent
IFN-gamma production by IFN-gamma ELISPOT assay, and the cells in
the positive well number #2 stimulated with KIF20A-A24-9-305 (SEQ
ID NO: 174), #3 with KIF20A-A24-9-383 (SEQ ID NO: 178), #5 with
KIF20A-A24-10-304 (SEQ ID NO: 186) and #6 with KIF20A-A24-10-66
(SEQ ID NO: 194) were expanded and CTL lines were established.
Those CTL lines having higher specific CTL activities against the
peptide-pulsed target as compared to the activities against target
without peptide pulse were determined by ELISA. Results are shown
in FIG. 6a-e. While, other peptides shown in table 7 could not
establish the CTL lines despite possible binding activity with
HLA-A*2402. For example, the typical negative peptide
(KIF20A-A24-9-647 and KIF20A-A24-10-182) were shown in FIG. 6a. In
this invention, the peptides which could establish CTL line were
selected as potent CTL stimulation peptide.
Establishment for CTL Clones Stimulated with KIF20A Derived
Peptides
[0477] Furthermore, the limiting dilution from these CTL lines was
performed according to the protocols set forth in the "Materials
and Methods" section above. The establishment of CTL clones from
KIF20A-A24-9-305 (SEQ ID NO: 174) #2 CTL line, KIF20A-A24-10-304
(SEQ ID NO: 186) #5 CTL line and KIF20A-A24-10-66 (SEQ ID NO: 194)
#6 CTL line are shown in FIGS. 6b, d and e. CTL clones had potent
and specific CTL activities against the peptide-pulsed target as
compared to the activities against target without peptide
pulse.
Specific CTL Activity Against the Target Cells Expressing KIF20A
and HLA-A*2402
[0478] The established CTL line raised against these peptides were
examined for their ability to recognize the target cells expressing
KIF20A and HLA-A*2402. Specific CTL activity against COS7
transfected with both full length KIF20A gene and the HLA-A*2402
molecule and A24-LCL transfected by electroporation with full
length KIF20A gene, which serve as a specific model for the target
cells endogenously express KIF20A and HLA-A*2402, was tested using
as effector cells the CTL lines raised by KIF20A-A24-9-383 (SEQ ID
NO: 178) and KIF20A-A24-10-304 (SEQ ID NO: 186) and the CTL clone
raised by KIF20A-A24-10-66 (SEQ ID NO: 194). COS7 transfected with
full length KIF20A but not HLA-A*2402 and COS7 transfected with
HLA-A*2402 but not full length KIF20A (or replaced full length
URLC10 gene), COS7 transfected with HLA-A*2402 and pulsed with
KIF20A-10-308, and A24-LCL transfected with mock vector were
prepared as controls. The CTL line demonstrated the highest
specific CTL activity against COS7 that was transfected with both
KIF20A and HLA-A*2402 (FIGS. 6b, c and d). Alternatively, the CTL
line stimulated with KIF20A-A24-10-304 (SEQ ID NO: 186)
demonstrated against A24-LCL transfected with KIF20A.
[0479] These results clearly demonstrate that KIF20A-A24-9-383 (SEQ
ID NO: 178), KIF20A-A24-10-304 (SEQ ID NO: 186) and
KIF20A-A24-10-66 (SEQ ID NO: 194) is naturally expressed on the
target cell surface with HLA-A2402 molecule and recognize CTL.
Furthermore, these peptides are epitope peptides, which may serve
as cancer vaccines targeting KIF20A expressed tumors.
Cytotoxic Activity Against Cancer Cell Line Endogenously Expressing
HLA-A*2402 and KIF20A
[0480] Furthermore, Cytotoxic activity was examined by cytotoxicity
assay according to the protocols set forth in the "Materials and
Methods" section above. As a result, as shown in FIGS. 6b and e,
CTL clone stimulated with KIF20A-A24-9-305 (SEQ ID NO: 174) or
KIF20A-A24-10-304 (SEQ ID NO: 186) showed remarkably high cytotoxic
effect towards HLA-A24-positive and KIF20A-positive cancer cell
lines PK45P or MIAPaca2 respectively, compared to that towards
HLA-A24-negative and KIF20A-positive cancer cell lines PK59.
Stimulation of the T Cells Using the Predicted Peptides from KNTC2
Restricted with HLA-A*2402, and Establishment for CTL Lines
Stimulated with KNTC2 Derived Peptides
[0481] CTLs for those peptides derived from KNTC2 were generated
according to the protocols set forth in the "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity as determined by an IFN-gamma ELISPOT assay are shown in
FIG. 7. In particular, KNTC2-A24-9-309 (SEQ ID NO: 196),
KNTC2-A24-9-124 (SEQ ID NO: 202), KNTC2-A24-9-154 (SEQ ID NO: 210),
KNTC2-A24-9-150 (SEQ ID NO: 213), KNTC2-A24-10-452 (SEQ ID NO:
214), KNTC2-A24-10-227 (SEQ ID NO: 217) and KNTC2-A24-10-273 (SEQ
ID NO: 223) demonstrated potent IFN-gamma production by IFN-gamma
ELISPOT assay, and the cells in the positive well number #8
stimulated with KNTC2-A24-9-309 (SEQ ID NO: 196), #5 with
KNTC2-A24-9-124 (SEQ ID NO: 202), #5 with KNTC2-A24-9-154 (SEQ ID
NO: 210), #7 with KNTC2-A24-9-150 (SEQ ID NO: 213), #4 and #5 with
KNTC2-A24-10-452 (SEQ ID NO: 214), #1 with KNTC2-A24-10-227 (SEQ ID
NO: 217) and #8 with KNTC2-A24-10-273 (SEQ ID NO: 223) were
expanded and CTL lines were established. Those CTL lines having
higher specific CTL activities against the peptide-pulsed target as
compared to the activities against target without peptide pulse
were determined by ELISA. Results are shown in FIG. 7a-h. While,
other peptides shown in table 8 could not establish the CTL lines
despite possible binding activity with HLA-A*2402. For example, the
typical negative peptide (KNTC2-A24-10-610) were shown in FIG. 7a.
In this invention, the peptides which could establish CTL line were
selected as potent CTL stimulation peptide.
Establishment for CTL Clones Stimulated with KNTC2 Derived
Peptides
[0482] Furthermore, the limiting dilution from these CTL lines was
performed according to the protocols set forth in the "Materials
and Methods" section above. The establishment of CTL clones from
KNTC2-A24-9-154 (SEQ ID NO: 210) #5 CTL line and KNTC2-A24-10-452
(SEQ ID NO: 214) #5 CTL line are shown in FIGS. 7d and f. CTL
clones had potent and specific CTL activities against the
peptide-pulsed target as compared to the activities against target
without peptide pulse.
Specific CTL Activity Against the Target Cells Expressing KNTC2 and
HLA-A*2402
[0483] The established CTL line raised against these peptides were
examined for their ability to recognize the target cells expressing
KNTC2 and HLA-A*2402. Specific CTL activity against HEK293
transfected with both full length KNTC2 gene and the HLA-A*2402
molecule which serves as a specific model for the target cells
endogenously express KNTC2 and HLA-A*2402, was tested using as
effector cells the CTL clones raised by KNTC2-A24-10-452 (SEQ ID
NO: 214). HEK293 transfected with full length KNTC2 but not
HLA-A*2402, HEK293 transfected with HLA-A*2402 but not full length
KNTC2 and HEK293 transfected with HLA-A*2402 and pulsed with
KNTC2-9-309 were prepared as controls. The CTL line demonstrating
the highest specific CTL activity against HEK293 was that
transfected with both KNTC2 and HLA-A*2402 (FIG. 7f).
[0484] These results clearly demonstrate that KNTC2-A24-10-452 (SEQ
ID NO: 214) is naturally expressed on the target cell surface with
HLA-A2402 molecule and recognize CTL. Furthermore, these peptides
are epitope peptides, which may serve as cancer vaccines targeting
KNTC2 expressed tumors.
Stimulation of the T Cells Using the Predicted Peptides from TTK
Restricted with HLA-A*0201, and Establishment for CTL Lines
Stimulated with TTK Derived Peptides
[0485] CTLs for those peptides derived from TTK were generated
according to the protocols set forth in the "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity as determined by an IFN-gamma ELISPOT assay are shown in
FIG. 8. As depicted in FIG. 8b-d, TTK-A2-9-462 (SEQ ID NO: 227),
TTK-A2-9-547 (SEQ ID NO: 228), TTK-A2-9-719 (SEQ ID NO: 233) and
TTK-A2-10-462 (SEQ ID NO: 254) demonstrated potent IFN-gamma
production by IFN-gamma ELISPOT assay, and the cells in the
positive well number #4 stimulated with TTK-A2-9-462 (SEQ ID NO:
227), #2 with TTK-A2-9-547 (SEQ ID NO: 228), #1 with TTK-A2-9-719
(SEQ ID NO: 233) and #8 with TTK-A2-10-462 (SEQ ID NO: 254) were
expanded. Those CTL lines having higher specific CTL activities
against the peptide-pulsed target as compared to the activities
against target without peptide pulse were determined by ELISA.
While, other peptides shown in table 9 could not establish the CTL
lines despite possible binding activity with HLA-A*0201. For
example, the typical negative peptide (TTK-A2-9-278) were shown in
FIG. 8a. In this invention, the peptides which could establish CTL
line were selected as potent CTL stimulation peptide.
Establishment for CTL Clones Stimulated with TTK Derived
Peptides
[0486] Furthermore, the limiting dilution from these CTL lines was
performed according to the protocols set forth in the "Materials
and Methods" section above. The establishment of CTL clones from
TTK-A2-9-462 (SEQ ID NO: 227) #4 CTL line, TTK-A2-9-547 (SEQ ID NO:
228) #2 CTL line, TTK-A2-9-719 (SEQ ID NO: 233) #1 CTL line and
TTK-A2-10-462 (SEQ ID NO: 254) #8 CTL line were shown in FIGS. 8d,
c, d and e. CTL clones had potent and specific CTL activities
against the peptide-pulsed target as compared to the activities
against target without peptide pulse.
Specific CTL Activity Against the Target Cells Expressing TTK and
HLA-A*0201
[0487] The established CTL clone raised against these peptides were
examined for their ability to recognize the target cells
endogenously expressing TTK and HLA-A*0201. Specific CTL activity
against COS7 transfected with both the full length TTK gene and the
HLA-A*0201 molecule, which is a specific model for the target cells
endogenously express TTK and HLA-A*0201, was tested using as
effector cells the CTL clones raised by TTK-A2-9-462 (SEQ ID NO:
227), TTK-A02-9-547 (SEQ ID NO: 228), TTK-A2-9-719 (SEQ ID NO: 233)
and TTK-A2-10-462 (SEQ ID NO: 254). COS7 transfected with full
length TTK but HLA-A*0201, COS7 transfected HLA-A*0201 but not full
length of TTK (or replaced full length HIG2 gene) and COS7
transfected with HLA-A*0201 and pulsed with different target
epitope peptide, were prepared as controls. The CTL Clone had the
highest specific CTL activity against COS7 that was transfected
with both TTK and HLA-A*0201 (FIGS. 8b, c, d and e).
[0488] These results clearly demonstrate that TTK-A2-9-462 (SEQ ID
NO: 227), TTK-A02-9-547 (SEQ ID NO: 228), TTK-A2-9-719 (SEQ ID NO:
233) and TTK-A02-10-462 (SEQ ID NO: 254) are naturally expressed on
the target cell surface with HLA-A2 (HLA-A02) molecule and
recognize CTL. Furthermore, these peptides are epitope peptides,
which may serve as cancer vaccines targeting TTK expressed
tumors.
Stimulation of the T Cells Using the Predicted Peptides from URLC10
Restricted with HLA-A*0201, and Establishment for CTL Lines
Stimulated with URLC10 Derived Peptides
[0489] CTLs for those peptides derived from URLC10 were generated
according to the protocols set forth in the "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity as determined by IFN-gamma ELISPOT assay are shown in FIG.
9. As shown in FIG. 9b-d, URLC-A2-9-206 (SEQ ID NO: 271),
URLC-A2-9-212 (SEQ ID NO: 272) and URLC-A2-10-211 (SEQ ID NO: 288)
demonstrated potent IFN-gamma production by IFN-gamma ELISPOT
assay, and the cells in the positive well number #7 stimulated with
URLC-A2-9-206 (SEQ ID NO: 271), #3 with URLC-A2-9-212 (SEQ ID NO:
272) and #5 with URLC-A2-10-211 (SEQ ID NO: 288) were expanded.
Those CTL lines having higher specific CTL activities against the
peptide-pulsed target as compared to the activities against target
without peptide pulse were determined by ELISA. While, other
peptides shown in table 10 could not establish the CTL lines
despite possible binding activity with HLA-A*0201. For example, the
typical negative peptide (URLC-A2-9-58) were shown in FIG. 9a. In
this invention, the peptide which could establish CTL line were
selected as potent CTL stimulation peptide.
Specific CTL Activity Against the Target Cells Expressing URLC10
and HLA-A*0201
[0490] The established CTL line raised against these peptides were
examined for their ability to recognize the target cells
endogenously expressing URLC10 and HLA-A*0201. Specific CTL
activity against COS7, Hek293 and 293T transfected with both full
length URLC10 gene and the HLA-A*0201 molecule, which serves as a
specific model for the target cells endogenously express URLC10 and
HLA-A*0201, was tested using as effector cells the CTL line raised
by URLC10-A02-10-211. COS7, Hek293 or 293T transfected with full
length URLC10 but not HLA-A*0201 (replaced HLA-A*2402), COS7,
Hek293 or 293T transfected with HLA-A*0201 but not full length
URLC10 and COS7 transfected with HLA-A*0201 and pulsed with
different target epitope peptide (URLC10-A02-10-64) were prepared
as controls. The CTL line demonstrating the highest specific CTL
activity against COS7, Hek293 or 293T was that transfected with
both URLC10 and HLA-A*0201 (FIG. 9-2).
[0491] These results clearly demonstrate that URLC10-A02-10-211 is
naturally expressed on the target cell surface with HLA-A*0201
molecule and recognizes CTL. Furthermore, this peptide was epitope
peptides, which may utilize cancer vaccine targeting URLC10
expressed tumors.
Homology Analysis of the Antigen Peptides
[0492] The CTL clones established against the following peptides
showed potent specific CTL activity. [0493] CDH3-A24-9-513 (SEQ ID
NO: 19), [0494] CDH3-A24-9-406 (SEQ ID NO: 22), [0495]
CDH3-A24-10-807 (SEQ ID NO: 30), [0496] CDH3-A24-10-332 (SEQ ID NO:
34), [0497] CDH3-A24-10-655 (SEQ ID NO: 344), [0498]
CDH3-A24-10-470 (SEQ ID NO: 358), [0499] EphA4-A24-9-453 (SEQ ID
NO: 41), [0500] EphA4-A24-9-5 (SEQ ID NO: 44), [0501]
EphA4-A24-9-869 (SEQ ID NO: 46), [0502] EphA4-A24-9-420 (SEQ ID NO:
48), [0503] EphA4-A24-10-24 (SEQ ID NO: 78), [0504] EphA4-A02-9-501
(SEQ ID NO: 376), [0505] EphA4-A02-9-165 (SEQ ID NO: 379), [0506]
HIG-A24-9-19 (SEQ ID NO: 110), [0507] HIG-A24-9-22 (SEQ ID NO:
111), [0508] HIG-A24-9-8 (SEQ ID NO: 387), [0509] HIG-A24-10-7 (SEQ
ID NO: 112), [0510] HIG-A24-10-18 (SEQ ID NO: 394), [0511]
HIG-A02-9-8 (SEQ ID NO: 114), [0512] HIG-A02-9-15 (SEQ ID NO: 116),
[0513] HIG-A02-9-4 (SEQ ID NO: 117), [0514] HIG-A02-10-8 (SEQ ID
NO: 121), [0515] INHBB-A24-9-180 (SEQ ID NO: 395), [0516]
INHBB-A24-10-180 (SEQ ID NO: 133), [0517] INHBB-A24-10-305 (SEQ ID
NO: 135), [0518] INHBB-A24-10-7 (SEQ ID NO: 137), [0519]
INHBB-A24-10-212 (SEQ ID NO: 426), [0520] KIF20A-A24-9-305 (SEQ ID
NO: 174), [0521] KIF20A-A24-9-383 (SEQ ID NO: 178), [0522]
KIF20A-A24-10-304 (SEQ ID NO: 186), [0523] KIF20A-A24-10-66 (SEQ ID
NO: 194), [0524] KNTC2-A24-9-309 (SEQ ID NO: 196), [0525]
KNTC2-A24-9-124 (SEQ ID NO: 202), [0526] KNTC2-A24-9-154 (SEQ ID
NO: 210), [0527] KNTC2-A24-9-150 (SEQ ID NO: 213), [0528]
KNTC2-A24-10-452 (SEQ ID NO: 214), [0529] KNTC2-A24-10-227 (SEQ ID
NO: 217), [0530] KNTC2-A24-10-273 (SEQ ID NO: 223), [0531]
TTK-A02-9-462 (SEQ ID NO: 227), [0532] TTK-A02-9-547 (SEQ ID NO:
228), [0533] TTK-A02-9-719 (SEQ ID NO: 233), [0534] TTK-A02-10-462
(SEQ ID NO: 254), [0535] URLC-A02-9-206 (SEQ ID NO: 271), [0536]
URLC-A02-9-212 (SEQ ID NO: 272) and [0537] URLC-A02-10-211 (SEQ ID
NO: 288)
[0538] This suggests that the sequences of SEQ ID NO: 19, 22, 30,
34, 344, 358, 41, 44, 46, 48, 78, 376, 379, 80, 100, 101, 110, 111,
387, 112, 394, 114, 116, 117, 121, 395, 133, 135, 137, 426, 174,
178, 186, 194, 196, 202, 210, 213, 214, 217, 223, 227, 228, 233,
254, 271, 272 or 288 are homologous to the peptides derived from
other molecules, which are known to sensitize human immune
system.
[0539] To exclude this possibility, homology analysis was performed
with the peptide sequences as queries using BLAST algorithm
(http://www.ncbi.nlm.nih.gov/blast/blast.cgi). No significant
sequence homology was revealed.
[0540] These results suggest that the sequences of SEQ ID NO: 19,
22, 30, 34, 344, 358, 41, 44, 46, 48, 78, 376, 379, 80, 100, 101,
110, 111, 387, 112, 394, 114, 116, 117, 121, 395, 133, 135, 137,
426, 174, 178, 186, 194, 196, 202, 210, 213, 214, 217, 223, 227,
228, 233, 254, 271, 272 or 288 are unique and thus possess a low
risk of raising unintended immunologic response to any unrelated
molecule.
Example 2
Materials and Methods
Cell Lines
[0541] H2 (HLA-A02), human B-lymphoblastoid cell line, and COS7
were purchased from ATCC.
Candidate Selection of Peptides Derived from INHBB
[0542] 9-mer and 10-mer peptides derived from INHBB that bind to
HLA-A*0201 molecules were predicted using binding prediction
software "BIMAS" (www-bimas.cit.nih.gov/molbio/hla_bind), which
algorithms had been described by Parker K C et al. (J Immunol 1994,
152(1): 163-75) and Kuzushima K et al. (Blood 2001, 98(6):
1872-81). These peptides were synthesized by Sigma (Sapporo, Japan)
or Biosynthesis Inc. (Lewisville, Tex.) according to a standard
solid phase synthesis method and purified by reversed phase high
performance liquid chromatography (HPLC). The purity (>90%) and
the identity of the peptides were determined by analytical HPLC and
mass spectrometry analysis, respectively. Peptides were dissolved
in dimethylsulfoxide (DMSO) at 20 mg/ml and stored at -80 degrees
C.
In Vitro CTL Induction
[0543] Monocyte-derived dendritic cells (DCs) were used as
antigen-presenting cells (APCs) to induce cytotoxic T lymphocyte
(CTL) responses against peptides presented on human leukocyte
antigen (HLA). DCs were generated in vitro as described elsewhere
(Nakahara S et al., Cancer Res 2003 Jul. 15, 63(14): 4112-8).
Specifically, peripheral blood mononuclear cells (PBMCs) isolated
from a normal volunteer (HLA-A*0201 positive) by Ficoll-Plaque
(Pharmacia) solution were separated by adherence to a plastic
tissue culture dish (Becton Dickinson) so as to enrich them as the
monocyte fraction. The monocyte-enriched population was cultured in
the presence of 1000 U/ml of granulocyte-macrophage
colony-stimulating factor (GM-CSF) (R&D System) and 1000 U/ml
of interleukin (IL)-4 (R&D System) in AIM-V Medium (Invitrogen)
containing 2% heat-inactivated autologous serum (AS). After 7 days
of culture, the cytokine-induced DCs were pulsed with 20 mcg/ml of
each of the synthesized peptides in the presence of 3 mcg/ml of
beta2-microglobulin for 3 hr at 37 degrees C. in AIM-V Medium. The
generated cells appeared to express DC-associated molecules, such
as CD80, CD83, CD86 and HLA class II, on their cell surfaces (data
not shown). These peptide-pulsed DCs were then inactivated by
Mitomycin C (MMC) (30 mcg/ml for 30 min) and mixed at a 1:20 ratio
with autologous CD8+ T cells, obtained by positive selection with
CD8 Positive Isolation Kit (Dynal). These cultures were set up in
48-well plates (Corning); each well contained 1.5.times.10.sup.4
peptide-pulsed DCs, 3.times.10.sup.5 CD8+ T cells and 10 ng/ml of
IL-7 (R&D System) in 0.5 ml of AIM-V/2% AS medium. Three days
later, these cultures were supplemented with IL-2 (CHIRON) to a
final concentration of 20 IU/ml. On day 7 and 14, the T cells were
further stimulated with the autologous peptide-pulsed DCs. The DCs
were prepared each time by the same way described above. CTL was
tested against peptide-pulsed T2 cells after the 3rd round of
peptide stimulation on day 21 (Tanaka H et al., Br J Cancer 2001
Jan. 5, 84(1): 94-9; Umano Y et al., Br J Cancer 2001 Apr. 20,
84(8): 1052-7; Uchida N et al., Clin Cancer Res 2004 Dec. 15,
10(24): 8577-86; Suda T et al., Cancer Sci 2006 May, 97(5): 411-9;
Watanabe T et al., Cancer Sci 2005 August, 96(8): 498-506).
CTL Expansion Procedure
[0544] CTLs were expanded in culture using the method similar to
the one described by Riddell et al. (Walter E A et al., N Engl J
Med 1995 Oct. 19, 333(16): 1038-44; Riddell S R et al., Nat Med
1996 Feb., 2(2): 216-23). A total of 5.times.10.sup.4 CTLs were
suspended in 25 ml of AIM-V/5% AS medium with 2 kinds of human
B-lymphoblastoid cell lines, inactivated by MMC, in the presence of
40 ng/ml of anti-CD3 monoclonal antibody (Pharmingen). One day
after initiating the cultures, 120 IU/ml of IL-2 were added to the
cultures. The cultures were fed with fresh AIM-V/5% AS medium
containing 30 IU/ml of IL-2 on days 5, 8 and 11 (Tanaka H et al.,
Br J Cancer 2001 Jan. 5, 84(1): 94-9; Umano Y et al., Br J Cancer
2001 Apr. 20, 84(8): 1052-7; Uchida N et. al., Clin Cancer Res 2004
Dec. 15, 10(24): 8577-86; Suda T et al., Cancer Sci 2006 May,
97(5): 411-9; Watanabe T et al., Cancer Sci 2005 August, 96(8):
498-506).
Specific CTL Activity
[0545] To examine specific CTL activity, interferon (IFN)-gamma
enzyme-linked immunospot (ELISPOT) assay and IFN-gamma
enzyme-linked immunosorbent assay (ELISA) were performed.
Specifically, peptide-pulsed T2 (1.times.10.sup.4/well) was
prepared as stimulator cells. Cultured cells in 48 wells were used
as responder cells. IFN-gamma ELISPOT assay and IFN-gamma ELISA
assay were performed under manufacture procedure.
Results
[0546] Stimulation of the T Cells Using the Predicted Peptides from
INHBB Restricted with HLA-A0201 and Establishment for CTL Lines
Stimulated with INHBB Derived Peptides
[0547] CTLs for those peptides derived from INHBB were generated
according to the protocols set forth in "Materials and Methods"
section above. Resulting CTLs having detectable specific CTL
activity, as determined by IFN-gamma ELISPOT assay, are shown in
FIG. 10. INHBB-A02-9-213 (SEQ ID NO: 143), INHBB-A02-9-174 (SEQ ID
NO: 147), INHBB-A02-9-257 (SEQ ID NO: 148), INHBB-A02-9-313 (SEQ ID
NO: 149), INHBB-A02-9-139 (SEQ ID NO: 150), INHBB-A02-9-8 (SEQ ID
NO: 152), INHBB-A02-9-250 (SEQ ID NO: 153), INHBB-A02-10-179 (SEQ
ID NO: 154), INHBB-A02-10-237 (SEQ ID NO: 156), INHBB-A02-10-313
(SEQ ID NO: 160), INHBB-A02-10-173 (SEQ ID NO: 161),
INHBB-A02-10-256 (SEQ ID NO: 162), INHBB-A02-10-162 (SEQ ID NO:
163) and INHBB-A02-10-85 (SEQ ID NO: 166) demonstrated potent
IFN-gamma production as compared to the control by IFN-gamma
ELISPOT assay. Furthermore, the cells in the positive well number
#7 stimulated with SEQ ID NO: 147, were expanded and CTL line was
established. The CTL line having higher specific CTL activity
against the peptide-pulsed target as compared to the activity
against target without peptide pulse was determined by IFN-gamma
ELISA (FIG. 11). The results herein demonstrate that the CTL line
demonstrated potent IFN-gamma production against the target cells
pulsed with corresponding peptide as compared to target cells
without peptide pulse. In the context of the present invention, the
peptides which could establish CTL line were selected as potent CTL
stimulation peptide.
[0548] In conclusion, novel HLA-A02 epitope peptides derived from
INHBB were identified and demonstrated to be applicable for cancer
immunotherapy.
DISCUSSION
[0549] Identification of new TAAs, particularly those that induce
potent and specific anti-tumor immune responses, warrants further
development of the clinical application of peptide vaccination
strategies in various types of cancer (Boon T. et al., (1996) J Exp
Med 183: 725-9.; van der Bruggen P et al., (1991) Science 254:
1643-7.; Brichard V et al., (1993) J Exp Med 178: 489-95.; Kawakami
Y et al., (1994) J Exp Med 180: 347-52.; Shichijo S et al., (1998)
J Exp Med 187:277-88.; Chen Y T et al., (1997) Proc. Natl. Acad.
Sci. USA, 94: 1914-8.; Harris C C., (1996) J Natl Cancer Inst
88:1442-5.; Butterfield L H et al., (1999) Cancer Res 59:3134-42.;
Vissers J L et al., (1999) Cancer Res 59: 5554-9.; van der Burg S H
et al., (1996) J. Immunol 156:3308-14.; Tanaka F et al., (1997)
Cancer Res 57:4465-8.; Fujie T et al., (1999) Int J Cancer
80:169-72.; Kikuchi M et al., (1999) Int J Cancer 81: 459-66.; Oiso
M et al., (1999) Int J Cancer 81:387-94.).
[0550] cDNA microarray technologies can disclose comprehensive
profiles of gene expression of malignant cells (Lin Y M, et al.,
Oncogene. 2002 Jun. 13; 21:4120-8.; Kitahara O, et al., Cancer Res.
2001 May 1; 61:3544-9.; Suzuki C, et al., Cancer Res. 2003 Nov. 1;
63:7038-41.; Ashida S, Cancer Res. 2004 Sep. 1; 64:5963-72.; Ochi
K, et al., Int J Oncol. 2004 March; 24(3):647-55.; Kaneta Y, et
al., Int J Oncol. 2003 September; 23:681-91.; Obama K, Hepatology.
2005 June; 41:1339-48.; Kato T, et al., Cancer Res. 2005 July 1;
65:5638-46.; Kitahara 0, et al., Neoplasia. 2002 July-August;
4:295-303.; Saito-Hisaminato A et al., DNA Res 2002, 9: 35-45.)
and, find utility in the identification of potential TAAs. Among
the transcripts that are up-regulated in various cancers, novel
human genes, termed CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2,
TTK and URLC10, were identified using these technologies.
[0551] As demonstrated above, CDH3, EPHA4, ECT2, HIG2, INHBB,
KIF20A, KNTC2, TTK and URLC10, are over-expressed in various
cancers but show minimal expression in normal tissues. In addition,
these genes have been shown to have a significant function related
to cell proliferation. Thus, peptides derived from CDH3, EPHA4,
ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK and URLC10 can serve as TAA
epitopes, which, in turn, can be used to induce significant and
specific immune responses against cancer cells.
[0552] Thus, as CDH3, EPHA4, ECT2, HIG2, INHBB, KIF20A, KNTC2, TTK
and URLC10 are novel TAAs, vaccines using these epitope peptides
find utility as immunotherapeutics against various carcinomas or
other disease expressing these molecules.
INDUSTRIAL APPLICABILITY
[0553] The present invention identifies new TAAs, particularly
those which induce potent and specific anti-tumor immune responses.
Such TAAs warrants further development as peptide vaccines against
diseases associated with the over-expression of CDH3, EPHA4, ECT2,
HIG2, INHBB, KIF20A, KNTC2, TTK and/or, URLC10 e.g. cancers. All
patents, patent applications, and publications cited herein are
incorporated by reference.
[0554] While the invention has been described in detail and with
reference to specific embodiments thereof, it is to be understood
that the foregoing description is exemplary and explanatory in
nature and is intended to illustrate the invention and its
preferred embodiments. Through routine experimentation, one skilled
in the art will readily recognize that various changes and
modifications can be made therein without departing from the spirit
and scope of the invention. Thus, the invention is intended to be
defined not by the above description, but by the following claims
and their equivalents.
[Sequence list]ONC-A0704Psq.txt
Sequence CWU 1
1
43613649DNAHomo sapiensCDS(508)..(2997) 1cccgctgtag ccgcgtgtgg
gaggacgcac gggcctgctt caaagctttg ggataacagc 60gcctccgggg gataatgaat
gcggagcctc cgttttcagt cgacttcaga tgtgtctcca 120cttttttccg
ctgtagccgc aaggcaagga aacatttctc ttcccgtact gaggaggctg
180aggagtgcac tgggtgttct tttctcctct aacccagaac tgcgagacag
aggctgagtc 240cctgtaaaga acagctccag aaaagccagg agagcgcagg
agggcatccg ggaggccagg 300aggggttcgc tggggcctca accgcaccca
catcggtccc acctgcgagg gggcgggacc 360tcgtggcgct ggaccaatca
gcacccacct gcgctcacct ggcctcctcc cgctggctcc 420cgggggctgc
ggtgctcaaa ggggcaagag ctgagcggaa caccggcccg ccgtcgcggc
480agctgcttca cccctctctc tgcagcc atg ggg ctc cct cgt gga cct ctc
gcg 534 Met Gly Leu Pro Arg Gly Pro Leu Ala 1 5 tct ctc ctc ctt ctc
cag gtt tgc tgg ctg cag tgc gcg gcc tcc gag 582Ser Leu Leu Leu Leu
Gln Val Cys Trp Leu Gln Cys Ala Ala Ser Glu 10 15 20 25 ccg tgc cgg
gcg gtc ttc agg gag gct gaa gtg acc ttg gag gcg gga 630Pro Cys Arg
Ala Val Phe Arg Glu Ala Glu Val Thr Leu Glu Ala Gly 30 35 40 ggc
gcg gag cag gag ccc ggc cag gcg ctg ggg aaa gta ttc atg ggc 678Gly
Ala Glu Gln Glu Pro Gly Gln Ala Leu Gly Lys Val Phe Met Gly 45 50
55 tgc cct ggg caa gag cca gct ctg ttt agc act gat aat gat gac ttc
726Cys Pro Gly Gln Glu Pro Ala Leu Phe Ser Thr Asp Asn Asp Asp Phe
60 65 70 act gtg cgg aat ggc gag aca gtc cag gaa aga agg tca ctg
aag gaa 774Thr Val Arg Asn Gly Glu Thr Val Gln Glu Arg Arg Ser Leu
Lys Glu 75 80 85 agg aat cca ttg aag atc ttc cca tcc aaa cgt atc
tta cga aga cac 822Arg Asn Pro Leu Lys Ile Phe Pro Ser Lys Arg Ile
Leu Arg Arg His 90 95 100 105 aag aga gat tgg gtg gtt gct cca ata
tct gtc cct gaa aat ggc aag 870Lys Arg Asp Trp Val Val Ala Pro Ile
Ser Val Pro Glu Asn Gly Lys 110 115 120 ggt ccc ttc ccc cag aga ctg
aat cag ctc aag tct aat aaa gat aga 918Gly Pro Phe Pro Gln Arg Leu
Asn Gln Leu Lys Ser Asn Lys Asp Arg 125 130 135 gac acc aag att ttc
tac agc atc acg ggg ccg ggg gca gac agc ccc 966Asp Thr Lys Ile Phe
Tyr Ser Ile Thr Gly Pro Gly Ala Asp Ser Pro 140 145 150 cct gag ggt
gtc ttc gct gta gag aag gag aca ggc tgg ttg ttg ttg 1014Pro Glu Gly
Val Phe Ala Val Glu Lys Glu Thr Gly Trp Leu Leu Leu 155 160 165 aat
aag cca ctg gac cgg gag gag att gcc aag tat gag ctc ttt ggc 1062Asn
Lys Pro Leu Asp Arg Glu Glu Ile Ala Lys Tyr Glu Leu Phe Gly 170 175
180 185 cac gct gtg tca gag aat ggt gcc tca gtg gag gac ccc atg aac
atc 1110His Ala Val Ser Glu Asn Gly Ala Ser Val Glu Asp Pro Met Asn
Ile 190 195 200 tcc atc atc gtg acc gac cag aat gac cac aag ccc aag
ttt acc cag 1158Ser Ile Ile Val Thr Asp Gln Asn Asp His Lys Pro Lys
Phe Thr Gln 205 210 215 gac acc ttc cga ggg agt gtc tta gag gga gtc
cta cca ggt act tct 1206Asp Thr Phe Arg Gly Ser Val Leu Glu Gly Val
Leu Pro Gly Thr Ser 220 225 230 gtg atg cag gtg aca gcc acg gat gag
gat gat gcc atc tac acc tac 1254Val Met Gln Val Thr Ala Thr Asp Glu
Asp Asp Ala Ile Tyr Thr Tyr 235 240 245 aat ggg gtg gtt gct tac tcc
atc cat agc caa gaa cca aag gac cca 1302Asn Gly Val Val Ala Tyr Ser
Ile His Ser Gln Glu Pro Lys Asp Pro 250 255 260 265 cac gac ctc atg
ttc acc att cac cgg agc aca ggc acc atc agc gtc 1350His Asp Leu Met
Phe Thr Ile His Arg Ser Thr Gly Thr Ile Ser Val 270 275 280 atc tcc
agt ggc ctg gac cgg gaa aaa gtc cct gag tac aca ctg acc 1398Ile Ser
Ser Gly Leu Asp Arg Glu Lys Val Pro Glu Tyr Thr Leu Thr 285 290 295
atc cag gcc aca gac atg gat ggg gac ggc tcc acc acc acg gca gtg
1446Ile Gln Ala Thr Asp Met Asp Gly Asp Gly Ser Thr Thr Thr Ala Val
300 305 310 gca gta gtg gag atc ctt gat gcc aat gac aat gct ccc atg
ttt gac 1494Ala Val Val Glu Ile Leu Asp Ala Asn Asp Asn Ala Pro Met
Phe Asp 315 320 325 ccc cag aag tac gag gcc cat gtg cct gag aat gca
gtg ggc cat gag 1542Pro Gln Lys Tyr Glu Ala His Val Pro Glu Asn Ala
Val Gly His Glu 330 335 340 345 gtg cag agg ctg acg gtc act gat ctg
gac gcc ccc aac tca cca gcg 1590Val Gln Arg Leu Thr Val Thr Asp Leu
Asp Ala Pro Asn Ser Pro Ala 350 355 360 tgg cgt gcc acc tac ctt atc
atg ggc ggt gac gac ggg gac cat ttt 1638Trp Arg Ala Thr Tyr Leu Ile
Met Gly Gly Asp Asp Gly Asp His Phe 365 370 375 acc atc acc acc cac
cct gag agc aac cag ggc atc ctg aca acc agg 1686Thr Ile Thr Thr His
Pro Glu Ser Asn Gln Gly Ile Leu Thr Thr Arg 380 385 390 aag ggt ttg
gat ttt gag gcc aaa aac cag cac acc ctg tac gtt gaa 1734Lys Gly Leu
Asp Phe Glu Ala Lys Asn Gln His Thr Leu Tyr Val Glu 395 400 405 gtg
acc aac gag gcc cct ttt gtg ctg aag ctc cca acc tcc aca gcc 1782Val
Thr Asn Glu Ala Pro Phe Val Leu Lys Leu Pro Thr Ser Thr Ala 410 415
420 425 acc ata gtg gtc cac gtg gag gat gtg aat gag gca cct gtg ttt
gtc 1830Thr Ile Val Val His Val Glu Asp Val Asn Glu Ala Pro Val Phe
Val 430 435 440 cca ccc tcc aaa gtc gtt gag gtc cag gag ggc atc ccc
act ggg gag 1878Pro Pro Ser Lys Val Val Glu Val Gln Glu Gly Ile Pro
Thr Gly Glu 445 450 455 cct gtg tgt gtc tac act gca gaa gac cct gac
aag gag aat caa aag 1926Pro Val Cys Val Tyr Thr Ala Glu Asp Pro Asp
Lys Glu Asn Gln Lys 460 465 470 atc agc tac cgc atc ctg aga gac cca
gca ggg tgg cta gcc atg gac 1974Ile Ser Tyr Arg Ile Leu Arg Asp Pro
Ala Gly Trp Leu Ala Met Asp 475 480 485 cca gac agt ggg cag gtc aca
gct gtg ggc acc ctc gac cgt gag gat 2022Pro Asp Ser Gly Gln Val Thr
Ala Val Gly Thr Leu Asp Arg Glu Asp 490 495 500 505 gag cag ttt gtg
agg aac aac atc tat gaa gtc atg gtc ttg gcc atg 2070Glu Gln Phe Val
Arg Asn Asn Ile Tyr Glu Val Met Val Leu Ala Met 510 515 520 gac aat
gga agc cct ccc acc act ggc acg gga acc ctt ctg cta aca 2118Asp Asn
Gly Ser Pro Pro Thr Thr Gly Thr Gly Thr Leu Leu Leu Thr 525 530 535
ctg att gat gtc aat gac cat ggc cca gtc cct gag ccc cgt cag atc
2166Leu Ile Asp Val Asn Asp His Gly Pro Val Pro Glu Pro Arg Gln Ile
540 545 550 acc atc tgc aac caa agc cct gtg cgc cag gtg ctg aac atc
acg gac 2214Thr Ile Cys Asn Gln Ser Pro Val Arg Gln Val Leu Asn Ile
Thr Asp 555 560 565 aag gac ctg tct ccc cac acc tcc cct ttc cag gcc
cag ctc aca gat 2262Lys Asp Leu Ser Pro His Thr Ser Pro Phe Gln Ala
Gln Leu Thr Asp 570 575 580 585 gac tca gac atc tac tgg acg gca gag
gtc aac gag gaa ggt gac aca 2310Asp Ser Asp Ile Tyr Trp Thr Ala Glu
Val Asn Glu Glu Gly Asp Thr 590 595 600 gtg gtc ttg tcc ctg aag aag
ttc ctg aag cag gat aca tat gac gtg 2358Val Val Leu Ser Leu Lys Lys
Phe Leu Lys Gln Asp Thr Tyr Asp Val 605 610 615 cac ctt tct ctg tct
gac cat ggc aac aaa gag cag ctg acg gtg atc 2406His Leu Ser Leu Ser
Asp His Gly Asn Lys Glu Gln Leu Thr Val Ile 620 625 630 agg gcc act
gtg tgc gac tgc cat ggc cat gtc gaa acc tgc cct gga 2454Arg Ala Thr
Val Cys Asp Cys His Gly His Val Glu Thr Cys Pro Gly 635 640 645 ccc
tgg aag gga ggt ttc atc ctc cct gtg ctg ggg gct gtc ctg gct 2502Pro
Trp Lys Gly Gly Phe Ile Leu Pro Val Leu Gly Ala Val Leu Ala 650 655
660 665 ctg ctg ttc ctc ctg ctg gtg ctg ctt ttg ttg gtg aga aag aag
cgg 2550Leu Leu Phe Leu Leu Leu Val Leu Leu Leu Leu Val Arg Lys Lys
Arg 670 675 680 aag atc aag gag ccc ctc cta ctc cca gaa gat gac acc
cgt gac aac 2598Lys Ile Lys Glu Pro Leu Leu Leu Pro Glu Asp Asp Thr
Arg Asp Asn 685 690 695 gtc ttc tac tat ggc gaa gag ggg ggt ggc gaa
gag gac cag gac tat 2646Val Phe Tyr Tyr Gly Glu Glu Gly Gly Gly Glu
Glu Asp Gln Asp Tyr 700 705 710 gac atc acc cag ctc cac cga ggt ctg
gag gcc agg ccg gag gtg gtt 2694Asp Ile Thr Gln Leu His Arg Gly Leu
Glu Ala Arg Pro Glu Val Val 715 720 725 ctc cgc aat gac gtg gca cca
acc atc atc ccg aca ccc atg tac cgt 2742Leu Arg Asn Asp Val Ala Pro
Thr Ile Ile Pro Thr Pro Met Tyr Arg 730 735 740 745 cct cgg cca gcc
aac cca gat gaa atc ggc aac ttt ata att gag aac 2790Pro Arg Pro Ala
Asn Pro Asp Glu Ile Gly Asn Phe Ile Ile Glu Asn 750 755 760 ctg aag
gcg gct aac aca gac ccc aca gcc ccg ccc tac gac acc ctc 2838Leu Lys
Ala Ala Asn Thr Asp Pro Thr Ala Pro Pro Tyr Asp Thr Leu 765 770 775
ttg gtg ttc gac tat gag ggc agc ggc tcc gac gcc gcg tcc ctg agc
2886Leu Val Phe Asp Tyr Glu Gly Ser Gly Ser Asp Ala Ala Ser Leu Ser
780 785 790 tcc ctc acc tcc tcc gcc tcc gac caa gac caa gat tac gat
tat ctg 2934Ser Leu Thr Ser Ser Ala Ser Asp Gln Asp Gln Asp Tyr Asp
Tyr Leu 795 800 805 aac gag tgg ggc agc cgc ttc aag aag ctg gca gac
atg tac ggt ggc 2982Asn Glu Trp Gly Ser Arg Phe Lys Lys Leu Ala Asp
Met Tyr Gly Gly 810 815 820 825 ggg gag gac gac tag gcggcctgcc
tgcagggctg gggaccaaac gtcaggccac 3037Gly Glu Asp Asp agagcatctc
caaggggtct cagttccccc ttcagctgag gacttcggag cttgtcagga
3097agtggccgta gcaacttggc ggagacaggc tatgagtctg acgttagagt
ggtggcttcc 3157ttagcctttc aggatggagg aatgtgggca gtttgacttc
agcactgaaa acctctccac 3217ctgggccagg gttgcctcag aggccaagtt
tccagaagcc tcttacctgc cgtaaaatgc 3277tcaaccctgt gtcctgggcc
tgggcctgct gtgactgacc tacagtggac tttctctctg 3337gaatggaacc
ttcttaggcc tcctggtgca acttaatttt tttttttaat gctatcttca
3397aaacgttaga gaaagttctt caaaagtgca gcccagagct gctgggccca
ctggccgtcc 3457tgcatttctg gtttccagac cccaatgcct cccattcgga
tggatctctg cgtttttata 3517ctgagtgtgc ctaggttgcc ccttattttt
tattttccct gttgcgttgc tatagatgaa 3577gggtgaggac aatcgtgtat
atgtactaga acttttttat taaagaaact tttcccagaa 3637aaaaaaaaaa aa
36492829PRTHomo sapiens 2Met Gly Leu Pro Arg Gly Pro Leu Ala Ser
Leu Leu Leu Leu Gln Val 1 5 10 15 Cys Trp Leu Gln Cys Ala Ala Ser
Glu Pro Cys Arg Ala Val Phe Arg 20 25 30 Glu Ala Glu Val Thr Leu
Glu Ala Gly Gly Ala Glu Gln Glu Pro Gly 35 40 45 Gln Ala Leu Gly
Lys Val Phe Met Gly Cys Pro Gly Gln Glu Pro Ala 50 55 60 Leu Phe
Ser Thr Asp Asn Asp Asp Phe Thr Val Arg Asn Gly Glu Thr 65 70 75 80
Val Gln Glu Arg Arg Ser Leu Lys Glu Arg Asn Pro Leu Lys Ile Phe 85
90 95 Pro Ser Lys Arg Ile Leu Arg Arg His Lys Arg Asp Trp Val Val
Ala 100 105 110 Pro Ile Ser Val Pro Glu Asn Gly Lys Gly Pro Phe Pro
Gln Arg Leu 115 120 125 Asn Gln Leu Lys Ser Asn Lys Asp Arg Asp Thr
Lys Ile Phe Tyr Ser 130 135 140 Ile Thr Gly Pro Gly Ala Asp Ser Pro
Pro Glu Gly Val Phe Ala Val 145 150 155 160 Glu Lys Glu Thr Gly Trp
Leu Leu Leu Asn Lys Pro Leu Asp Arg Glu 165 170 175 Glu Ile Ala Lys
Tyr Glu Leu Phe Gly His Ala Val Ser Glu Asn Gly 180 185 190 Ala Ser
Val Glu Asp Pro Met Asn Ile Ser Ile Ile Val Thr Asp Gln 195 200 205
Asn Asp His Lys Pro Lys Phe Thr Gln Asp Thr Phe Arg Gly Ser Val 210
215 220 Leu Glu Gly Val Leu Pro Gly Thr Ser Val Met Gln Val Thr Ala
Thr 225 230 235 240 Asp Glu Asp Asp Ala Ile Tyr Thr Tyr Asn Gly Val
Val Ala Tyr Ser 245 250 255 Ile His Ser Gln Glu Pro Lys Asp Pro His
Asp Leu Met Phe Thr Ile 260 265 270 His Arg Ser Thr Gly Thr Ile Ser
Val Ile Ser Ser Gly Leu Asp Arg 275 280 285 Glu Lys Val Pro Glu Tyr
Thr Leu Thr Ile Gln Ala Thr Asp Met Asp 290 295 300 Gly Asp Gly Ser
Thr Thr Thr Ala Val Ala Val Val Glu Ile Leu Asp 305 310 315 320 Ala
Asn Asp Asn Ala Pro Met Phe Asp Pro Gln Lys Tyr Glu Ala His 325 330
335 Val Pro Glu Asn Ala Val Gly His Glu Val Gln Arg Leu Thr Val Thr
340 345 350 Asp Leu Asp Ala Pro Asn Ser Pro Ala Trp Arg Ala Thr Tyr
Leu Ile 355 360 365 Met Gly Gly Asp Asp Gly Asp His Phe Thr Ile Thr
Thr His Pro Glu 370 375 380 Ser Asn Gln Gly Ile Leu Thr Thr Arg Lys
Gly Leu Asp Phe Glu Ala 385 390 395 400 Lys Asn Gln His Thr Leu Tyr
Val Glu Val Thr Asn Glu Ala Pro Phe 405 410 415 Val Leu Lys Leu Pro
Thr Ser Thr Ala Thr Ile Val Val His Val Glu 420 425 430 Asp Val Asn
Glu Ala Pro Val Phe Val Pro Pro Ser Lys Val Val Glu 435 440 445 Val
Gln Glu Gly Ile Pro Thr Gly Glu Pro Val Cys Val Tyr Thr Ala 450 455
460 Glu Asp Pro Asp Lys Glu Asn Gln Lys Ile Ser Tyr Arg Ile Leu Arg
465 470 475 480 Asp Pro Ala Gly Trp Leu Ala Met Asp Pro Asp Ser Gly
Gln Val Thr 485 490 495 Ala Val Gly Thr Leu Asp Arg Glu Asp Glu Gln
Phe Val Arg Asn Asn 500 505 510 Ile Tyr Glu Val Met Val Leu Ala Met
Asp Asn Gly Ser Pro Pro Thr 515 520 525 Thr Gly Thr Gly Thr Leu Leu
Leu Thr Leu Ile Asp Val Asn Asp His 530 535 540 Gly Pro Val Pro Glu
Pro Arg Gln Ile Thr Ile Cys Asn Gln Ser Pro 545 550 555 560 Val Arg
Gln Val Leu Asn Ile Thr Asp Lys Asp Leu Ser Pro His Thr 565 570 575
Ser Pro Phe Gln Ala Gln Leu Thr Asp Asp Ser Asp Ile Tyr Trp Thr 580
585 590 Ala Glu Val Asn Glu Glu Gly Asp Thr Val Val Leu Ser Leu Lys
Lys 595 600 605 Phe Leu Lys Gln Asp Thr Tyr Asp Val His Leu Ser Leu
Ser Asp His 610 615 620 Gly Asn Lys Glu Gln Leu Thr Val Ile Arg Ala
Thr Val Cys Asp Cys 625 630 635 640 His Gly His Val Glu Thr Cys Pro
Gly Pro Trp Lys Gly Gly Phe Ile 645 650 655 Leu Pro Val Leu Gly Ala
Val Leu Ala Leu Leu Phe Leu Leu Leu Val 660 665 670 Leu Leu Leu Leu
Val Arg Lys Lys Arg Lys Ile Lys Glu
Pro Leu Leu 675 680 685 Leu Pro Glu Asp Asp Thr Arg Asp Asn Val Phe
Tyr Tyr Gly Glu Glu 690 695 700 Gly Gly Gly Glu Glu Asp Gln Asp Tyr
Asp Ile Thr Gln Leu His Arg 705 710 715 720 Gly Leu Glu Ala Arg Pro
Glu Val Val Leu Arg Asn Asp Val Ala Pro 725 730 735 Thr Ile Ile Pro
Thr Pro Met Tyr Arg Pro Arg Pro Ala Asn Pro Asp 740 745 750 Glu Ile
Gly Asn Phe Ile Ile Glu Asn Leu Lys Ala Ala Asn Thr Asp 755 760 765
Pro Thr Ala Pro Pro Tyr Asp Thr Leu Leu Val Phe Asp Tyr Glu Gly 770
775 780 Ser Gly Ser Asp Ala Ala Ser Leu Ser Ser Leu Thr Ser Ser Ala
Ser 785 790 795 800 Asp Gln Asp Gln Asp Tyr Asp Tyr Leu Asn Glu Trp
Gly Ser Arg Phe 805 810 815 Lys Lys Leu Ala Asp Met Tyr Gly Gly Gly
Glu Asp Asp 820 825 33107DNAHomo sapiensCDS(34)..(2994) 3aagcggcagg
agcagcgttg gcaccggcga acc atg gct ggg att ttc tat ttc 54 Met Ala
Gly Ile Phe Tyr Phe 1 5 gcc cta ttt tcg tgt ctc ttc ggg att tgc gac
gct gtc aca ggt tcc 102Ala Leu Phe Ser Cys Leu Phe Gly Ile Cys Asp
Ala Val Thr Gly Ser 10 15 20 agg gta tac ccc gcg aat gaa gtt acc
tta ttg gat tcc aga tct gtt 150Arg Val Tyr Pro Ala Asn Glu Val Thr
Leu Leu Asp Ser Arg Ser Val 25 30 35 cag gga gaa ctt ggg tgg ata
gca agc cct ctg gaa gga ggg tgg gag 198Gln Gly Glu Leu Gly Trp Ile
Ala Ser Pro Leu Glu Gly Gly Trp Glu 40 45 50 55 gaa gtg agt atc atg
gat gaa aaa aat aca cca atc cga acc tac caa 246Glu Val Ser Ile Met
Asp Glu Lys Asn Thr Pro Ile Arg Thr Tyr Gln 60 65 70 gtg tgc aat
gtg atg gaa ccc agc cag aat aac tgg cta cga act gat 294Val Cys Asn
Val Met Glu Pro Ser Gln Asn Asn Trp Leu Arg Thr Asp 75 80 85 tgg
atc acc cga gaa ggg gct cag agg gtg tat att gag att aaa ttc 342Trp
Ile Thr Arg Glu Gly Ala Gln Arg Val Tyr Ile Glu Ile Lys Phe 90 95
100 acc ttg agg gac tgc aat agt ctt ccg ggc gtc atg ggg act tgc aag
390Thr Leu Arg Asp Cys Asn Ser Leu Pro Gly Val Met Gly Thr Cys Lys
105 110 115 gag acg ttt aac ctg tac tac tat gaa tca gac aac gac aaa
gag cgt 438Glu Thr Phe Asn Leu Tyr Tyr Tyr Glu Ser Asp Asn Asp Lys
Glu Arg 120 125 130 135 ttc atc aga gag aac cag ttt gtc aaa att gac
acc att gct gct gat 486Phe Ile Arg Glu Asn Gln Phe Val Lys Ile Asp
Thr Ile Ala Ala Asp 140 145 150 gag agc ttc acc caa gtg gac att ggt
gac aga atc atg aag ctg aac 534Glu Ser Phe Thr Gln Val Asp Ile Gly
Asp Arg Ile Met Lys Leu Asn 155 160 165 acc gag atc cgg gat gta ggg
cca tta agc aaa aag ggg ttt tac ctg 582Thr Glu Ile Arg Asp Val Gly
Pro Leu Ser Lys Lys Gly Phe Tyr Leu 170 175 180 gct ttt cag gat gtg
ggg gcc tgc atc gcc ctg gta tca gtc cgt gtg 630Ala Phe Gln Asp Val
Gly Ala Cys Ile Ala Leu Val Ser Val Arg Val 185 190 195 ttc tat aaa
aag tgt cca ctc aca gtc cgc aat ctg gcc cag ttt cct 678Phe Tyr Lys
Lys Cys Pro Leu Thr Val Arg Asn Leu Ala Gln Phe Pro 200 205 210 215
gac acc atc aca ggg gct gat acg tct tcc ctg gtg gaa gtt cga ggc
726Asp Thr Ile Thr Gly Ala Asp Thr Ser Ser Leu Val Glu Val Arg Gly
220 225 230 tcc tgt gtc aac aac tca gaa gag aaa gat gtg cca aaa atg
tac tgt 774Ser Cys Val Asn Asn Ser Glu Glu Lys Asp Val Pro Lys Met
Tyr Cys 235 240 245 ggg gca gat ggt gaa tgg ctg gta ccc att ggc aac
tgc cta tgc aac 822Gly Ala Asp Gly Glu Trp Leu Val Pro Ile Gly Asn
Cys Leu Cys Asn 250 255 260 gct ggg cat gag gag cgg agc gga gaa tgc
caa gct tgc aaa att gga 870Ala Gly His Glu Glu Arg Ser Gly Glu Cys
Gln Ala Cys Lys Ile Gly 265 270 275 tat tac aag gct ctc tcc acg gat
gcc acc tgt gcc aag tgc cca ccc 918Tyr Tyr Lys Ala Leu Ser Thr Asp
Ala Thr Cys Ala Lys Cys Pro Pro 280 285 290 295 cac agc tac tct gtc
tgg gaa gga gcc acc tcg tgc acc tgt gac cga 966His Ser Tyr Ser Val
Trp Glu Gly Ala Thr Ser Cys Thr Cys Asp Arg 300 305 310 ggc ttt ttc
aga gct gac aac gat gct gcc tct atg ccc tgc acc cgt 1014Gly Phe Phe
Arg Ala Asp Asn Asp Ala Ala Ser Met Pro Cys Thr Arg 315 320 325 cca
cca tct gct ccc ctg aac ttg att tca aat gtc aac gag aca tct 1062Pro
Pro Ser Ala Pro Leu Asn Leu Ile Ser Asn Val Asn Glu Thr Ser 330 335
340 gtg aac ttg gaa tgg agt agc cct cag aat aca ggt ggc cgc cag gac
1110Val Asn Leu Glu Trp Ser Ser Pro Gln Asn Thr Gly Gly Arg Gln Asp
345 350 355 att tcc tat aat gtg gta tgc aag aaa tgt gga gct ggt gac
ccc agc 1158Ile Ser Tyr Asn Val Val Cys Lys Lys Cys Gly Ala Gly Asp
Pro Ser 360 365 370 375 aag tgc cga ccc tgt gga agt ggg gtc cac tac
acc cca cag cag aat 1206Lys Cys Arg Pro Cys Gly Ser Gly Val His Tyr
Thr Pro Gln Gln Asn 380 385 390 ggc ttg aag acc acc aaa gtc tcc atc
act gac ctc cta gct cat acc 1254Gly Leu Lys Thr Thr Lys Val Ser Ile
Thr Asp Leu Leu Ala His Thr 395 400 405 aat tac acc ttt gaa atc tgg
gct gtg aat gga gtg tcc aaa tat aac 1302Asn Tyr Thr Phe Glu Ile Trp
Ala Val Asn Gly Val Ser Lys Tyr Asn 410 415 420 cct aac cca gac caa
tca gtt tct gtc act gtg acc acc aac caa gca 1350Pro Asn Pro Asp Gln
Ser Val Ser Val Thr Val Thr Thr Asn Gln Ala 425 430 435 gca cca tca
tcc att gct ttg gtc cag gct aaa gaa gtc aca aga tac 1398Ala Pro Ser
Ser Ile Ala Leu Val Gln Ala Lys Glu Val Thr Arg Tyr 440 445 450 455
agt gtg gca ctg gct tgg ctg gaa cca gat cgg ccc aat ggg gta atc
1446Ser Val Ala Leu Ala Trp Leu Glu Pro Asp Arg Pro Asn Gly Val Ile
460 465 470 ctg gaa tat gaa gtc aag tat tat gag aag gat cag aat gag
cga agc 1494Leu Glu Tyr Glu Val Lys Tyr Tyr Glu Lys Asp Gln Asn Glu
Arg Ser 475 480 485 tat cgt ata gtt cgg aca gct gcc agg aac aca gat
atc aaa ggc ctg 1542Tyr Arg Ile Val Arg Thr Ala Ala Arg Asn Thr Asp
Ile Lys Gly Leu 490 495 500 aac cct ctc act tcc tat gtt ttc cac gtg
cga gcc agg aca gca gct 1590Asn Pro Leu Thr Ser Tyr Val Phe His Val
Arg Ala Arg Thr Ala Ala 505 510 515 ggc tat gga gac ttc agt gag ccc
ttg gag gtt aca acc aac aca gtg 1638Gly Tyr Gly Asp Phe Ser Glu Pro
Leu Glu Val Thr Thr Asn Thr Val 520 525 530 535 cct tcc cgg atc att
gga gat ggg gct aac tcc aca gtc ctt ctg gtc 1686Pro Ser Arg Ile Ile
Gly Asp Gly Ala Asn Ser Thr Val Leu Leu Val 540 545 550 tct gtc tcg
ggc agt gtg gtg ctg gtg gta att ctc att gca gct ttt 1734Ser Val Ser
Gly Ser Val Val Leu Val Val Ile Leu Ile Ala Ala Phe 555 560 565 gtc
atc agc cgg aga cgg agt aaa tac agt aaa gcc aaa caa gaa gcg 1782Val
Ile Ser Arg Arg Arg Ser Lys Tyr Ser Lys Ala Lys Gln Glu Ala 570 575
580 gat gaa gag aaa cat ttg aat caa ggt gta aga aca tat gtg gac ccc
1830Asp Glu Glu Lys His Leu Asn Gln Gly Val Arg Thr Tyr Val Asp Pro
585 590 595 ttt acg tac gaa gat ccc aac caa gca gtg cga gag ttt gcc
aaa gaa 1878Phe Thr Tyr Glu Asp Pro Asn Gln Ala Val Arg Glu Phe Ala
Lys Glu 600 605 610 615 att gac gca tcc tgc att aag att gaa aaa gtt
ata gga gtt ggt gaa 1926Ile Asp Ala Ser Cys Ile Lys Ile Glu Lys Val
Ile Gly Val Gly Glu 620 625 630 ttt ggt gag gta tgc agt ggg cgt ctc
aaa gtg cct ggc aag aga gag 1974Phe Gly Glu Val Cys Ser Gly Arg Leu
Lys Val Pro Gly Lys Arg Glu 635 640 645 atc tgt gtg gct atc aag act
ctg aaa gct ggt tat aca gac aaa cag 2022Ile Cys Val Ala Ile Lys Thr
Leu Lys Ala Gly Tyr Thr Asp Lys Gln 650 655 660 agg aga gac ttc ctg
agt gag gcc agc atc atg gga cag ttt gac cat 2070Arg Arg Asp Phe Leu
Ser Glu Ala Ser Ile Met Gly Gln Phe Asp His 665 670 675 ccg aac atc
att cac ttg gaa ggc gtg gtc act aaa tgt aaa cca gta 2118Pro Asn Ile
Ile His Leu Glu Gly Val Val Thr Lys Cys Lys Pro Val 680 685 690 695
atg atc ata aca gag tac atg gag aat ggc tcc ttg gat gca ttc ctc
2166Met Ile Ile Thr Glu Tyr Met Glu Asn Gly Ser Leu Asp Ala Phe Leu
700 705 710 agg aaa aat gat ggc aga ttt aca gtc att cag ctg gtg ggc
atg ctt 2214Arg Lys Asn Asp Gly Arg Phe Thr Val Ile Gln Leu Val Gly
Met Leu 715 720 725 cgt ggc att ggg tct ggg atg aag tat tta tct gat
atg agc tat gtg 2262Arg Gly Ile Gly Ser Gly Met Lys Tyr Leu Ser Asp
Met Ser Tyr Val 730 735 740 cat cgt gat ctg gcc gca cgg aac atc ctg
gtg aac agc aac ttg gtc 2310His Arg Asp Leu Ala Ala Arg Asn Ile Leu
Val Asn Ser Asn Leu Val 745 750 755 tgc aaa gtg tct gat ttt ggc atg
tcc cga gtg ctt gag gat gat ccg 2358Cys Lys Val Ser Asp Phe Gly Met
Ser Arg Val Leu Glu Asp Asp Pro 760 765 770 775 gaa gca gct tac acc
acc agg ggt ggc aag att cct atc cgg tgg act 2406Glu Ala Ala Tyr Thr
Thr Arg Gly Gly Lys Ile Pro Ile Arg Trp Thr 780 785 790 gcg cca gaa
gca att gcc tat cgt aaa ttc aca tca gca agt gat gta 2454Ala Pro Glu
Ala Ile Ala Tyr Arg Lys Phe Thr Ser Ala Ser Asp Val 795 800 805 tgg
agc tat gga atc gtt atg tgg gaa gtg atg tcg tac ggg gag agg 2502Trp
Ser Tyr Gly Ile Val Met Trp Glu Val Met Ser Tyr Gly Glu Arg 810 815
820 ccc tat tgg gat atg tcc aat caa gat gtg att aaa gcc att gag gaa
2550Pro Tyr Trp Asp Met Ser Asn Gln Asp Val Ile Lys Ala Ile Glu Glu
825 830 835 ggc tat cgg tta ccc cct cca atg gac tgc ccc att gcg ctc
cac cag 2598Gly Tyr Arg Leu Pro Pro Pro Met Asp Cys Pro Ile Ala Leu
His Gln 840 845 850 855 ctg atg cta gac tgc tgg cag aag gag agg agc
gac agg cct aaa ttt 2646Leu Met Leu Asp Cys Trp Gln Lys Glu Arg Ser
Asp Arg Pro Lys Phe 860 865 870 ggg cag att gtc aac atg ttg gac aaa
ctc atc cgc aac ccc aac agc 2694Gly Gln Ile Val Asn Met Leu Asp Lys
Leu Ile Arg Asn Pro Asn Ser 875 880 885 ttg aag agg aca ggg acg gag
agc tcc aga cct aac act gcc ttg ttg 2742Leu Lys Arg Thr Gly Thr Glu
Ser Ser Arg Pro Asn Thr Ala Leu Leu 890 895 900 gat cca agc tcc cct
gaa ttc tct gct gtg gta tca gtg ggc gat tgg 2790Asp Pro Ser Ser Pro
Glu Phe Ser Ala Val Val Ser Val Gly Asp Trp 905 910 915 ctc cag gcc
att aaa atg gac cgg tat aag gat aac ttc aca gct gct 2838Leu Gln Ala
Ile Lys Met Asp Arg Tyr Lys Asp Asn Phe Thr Ala Ala 920 925 930 935
ggt tat acc aca cta gag gct gtg gtg cac gtg aac cag gag gac ctg
2886Gly Tyr Thr Thr Leu Glu Ala Val Val His Val Asn Gln Glu Asp Leu
940 945 950 gca aga att ggt atc aca gcc atc acg cac cag aat aag att
ttg agc 2934Ala Arg Ile Gly Ile Thr Ala Ile Thr His Gln Asn Lys Ile
Leu Ser 955 960 965 agt gtc cag gca atg cga acc caa atg cag cag atg
cac ggc aga atg 2982Ser Val Gln Ala Met Arg Thr Gln Met Gln Gln Met
His Gly Arg Met 970 975 980 gtt ccc gtc tga gccagtactg aataaactca
aaactcttga aattagttta 3034Val Pro Val 985 cctcatccat gcactttaat
tgaagaactg cacttttttt acttcgtctt cgccctctga 3094aattaaagaa atg
31074986PRTHomo sapiens 4Met Ala Gly Ile Phe Tyr Phe Ala Leu Phe
Ser Cys Leu Phe Gly Ile 1 5 10 15 Cys Asp Ala Val Thr Gly Ser Arg
Val Tyr Pro Ala Asn Glu Val Thr 20 25 30 Leu Leu Asp Ser Arg Ser
Val Gln Gly Glu Leu Gly Trp Ile Ala Ser 35 40 45 Pro Leu Glu Gly
Gly Trp Glu Glu Val Ser Ile Met Asp Glu Lys Asn 50 55 60 Thr Pro
Ile Arg Thr Tyr Gln Val Cys Asn Val Met Glu Pro Ser Gln 65 70 75 80
Asn Asn Trp Leu Arg Thr Asp Trp Ile Thr Arg Glu Gly Ala Gln Arg 85
90 95 Val Tyr Ile Glu Ile Lys Phe Thr Leu Arg Asp Cys Asn Ser Leu
Pro 100 105 110 Gly Val Met Gly Thr Cys Lys Glu Thr Phe Asn Leu Tyr
Tyr Tyr Glu 115 120 125 Ser Asp Asn Asp Lys Glu Arg Phe Ile Arg Glu
Asn Gln Phe Val Lys 130 135 140 Ile Asp Thr Ile Ala Ala Asp Glu Ser
Phe Thr Gln Val Asp Ile Gly 145 150 155 160 Asp Arg Ile Met Lys Leu
Asn Thr Glu Ile Arg Asp Val Gly Pro Leu 165 170 175 Ser Lys Lys Gly
Phe Tyr Leu Ala Phe Gln Asp Val Gly Ala Cys Ile 180 185 190 Ala Leu
Val Ser Val Arg Val Phe Tyr Lys Lys Cys Pro Leu Thr Val 195 200 205
Arg Asn Leu Ala Gln Phe Pro Asp Thr Ile Thr Gly Ala Asp Thr Ser 210
215 220 Ser Leu Val Glu Val Arg Gly Ser Cys Val Asn Asn Ser Glu Glu
Lys 225 230 235 240 Asp Val Pro Lys Met Tyr Cys Gly Ala Asp Gly Glu
Trp Leu Val Pro 245 250 255 Ile Gly Asn Cys Leu Cys Asn Ala Gly His
Glu Glu Arg Ser Gly Glu 260 265 270 Cys Gln Ala Cys Lys Ile Gly Tyr
Tyr Lys Ala Leu Ser Thr Asp Ala 275 280 285 Thr Cys Ala Lys Cys Pro
Pro His Ser Tyr Ser Val Trp Glu Gly Ala 290 295 300 Thr Ser Cys Thr
Cys Asp Arg Gly Phe Phe Arg Ala Asp Asn Asp Ala 305 310 315 320 Ala
Ser Met Pro Cys Thr Arg Pro Pro Ser Ala Pro Leu Asn Leu Ile 325 330
335 Ser Asn Val Asn Glu Thr Ser Val Asn Leu Glu Trp Ser Ser Pro Gln
340 345 350 Asn Thr Gly Gly Arg Gln Asp Ile Ser Tyr Asn Val Val Cys
Lys Lys 355 360 365 Cys Gly Ala Gly Asp Pro Ser Lys Cys Arg Pro Cys
Gly Ser Gly Val 370 375 380 His Tyr Thr Pro Gln Gln Asn Gly Leu Lys
Thr Thr Lys Val Ser Ile 385
390 395 400 Thr Asp Leu Leu Ala His Thr Asn Tyr Thr Phe Glu Ile Trp
Ala Val 405 410 415 Asn Gly Val Ser Lys Tyr Asn Pro Asn Pro Asp Gln
Ser Val Ser Val 420 425 430 Thr Val Thr Thr Asn Gln Ala Ala Pro Ser
Ser Ile Ala Leu Val Gln 435 440 445 Ala Lys Glu Val Thr Arg Tyr Ser
Val Ala Leu Ala Trp Leu Glu Pro 450 455 460 Asp Arg Pro Asn Gly Val
Ile Leu Glu Tyr Glu Val Lys Tyr Tyr Glu 465 470 475 480 Lys Asp Gln
Asn Glu Arg Ser Tyr Arg Ile Val Arg Thr Ala Ala Arg 485 490 495 Asn
Thr Asp Ile Lys Gly Leu Asn Pro Leu Thr Ser Tyr Val Phe His 500 505
510 Val Arg Ala Arg Thr Ala Ala Gly Tyr Gly Asp Phe Ser Glu Pro Leu
515 520 525 Glu Val Thr Thr Asn Thr Val Pro Ser Arg Ile Ile Gly Asp
Gly Ala 530 535 540 Asn Ser Thr Val Leu Leu Val Ser Val Ser Gly Ser
Val Val Leu Val 545 550 555 560 Val Ile Leu Ile Ala Ala Phe Val Ile
Ser Arg Arg Arg Ser Lys Tyr 565 570 575 Ser Lys Ala Lys Gln Glu Ala
Asp Glu Glu Lys His Leu Asn Gln Gly 580 585 590 Val Arg Thr Tyr Val
Asp Pro Phe Thr Tyr Glu Asp Pro Asn Gln Ala 595 600 605 Val Arg Glu
Phe Ala Lys Glu Ile Asp Ala Ser Cys Ile Lys Ile Glu 610 615 620 Lys
Val Ile Gly Val Gly Glu Phe Gly Glu Val Cys Ser Gly Arg Leu 625 630
635 640 Lys Val Pro Gly Lys Arg Glu Ile Cys Val Ala Ile Lys Thr Leu
Lys 645 650 655 Ala Gly Tyr Thr Asp Lys Gln Arg Arg Asp Phe Leu Ser
Glu Ala Ser 660 665 670 Ile Met Gly Gln Phe Asp His Pro Asn Ile Ile
His Leu Glu Gly Val 675 680 685 Val Thr Lys Cys Lys Pro Val Met Ile
Ile Thr Glu Tyr Met Glu Asn 690 695 700 Gly Ser Leu Asp Ala Phe Leu
Arg Lys Asn Asp Gly Arg Phe Thr Val 705 710 715 720 Ile Gln Leu Val
Gly Met Leu Arg Gly Ile Gly Ser Gly Met Lys Tyr 725 730 735 Leu Ser
Asp Met Ser Tyr Val His Arg Asp Leu Ala Ala Arg Asn Ile 740 745 750
Leu Val Asn Ser Asn Leu Val Cys Lys Val Ser Asp Phe Gly Met Ser 755
760 765 Arg Val Leu Glu Asp Asp Pro Glu Ala Ala Tyr Thr Thr Arg Gly
Gly 770 775 780 Lys Ile Pro Ile Arg Trp Thr Ala Pro Glu Ala Ile Ala
Tyr Arg Lys 785 790 795 800 Phe Thr Ser Ala Ser Asp Val Trp Ser Tyr
Gly Ile Val Met Trp Glu 805 810 815 Val Met Ser Tyr Gly Glu Arg Pro
Tyr Trp Asp Met Ser Asn Gln Asp 820 825 830 Val Ile Lys Ala Ile Glu
Glu Gly Tyr Arg Leu Pro Pro Pro Met Asp 835 840 845 Cys Pro Ile Ala
Leu His Gln Leu Met Leu Asp Cys Trp Gln Lys Glu 850 855 860 Arg Ser
Asp Arg Pro Lys Phe Gly Gln Ile Val Asn Met Leu Asp Lys 865 870 875
880 Leu Ile Arg Asn Pro Asn Ser Leu Lys Arg Thr Gly Thr Glu Ser Ser
885 890 895 Arg Pro Asn Thr Ala Leu Leu Asp Pro Ser Ser Pro Glu Phe
Ser Ala 900 905 910 Val Val Ser Val Gly Asp Trp Leu Gln Ala Ile Lys
Met Asp Arg Tyr 915 920 925 Lys Asp Asn Phe Thr Ala Ala Gly Tyr Thr
Thr Leu Glu Ala Val Val 930 935 940 His Val Asn Gln Glu Asp Leu Ala
Arg Ile Gly Ile Thr Ala Ile Thr 945 950 955 960 His Gln Asn Lys Ile
Leu Ser Ser Val Gln Ala Met Arg Thr Gln Met 965 970 975 Gln Gln Met
His Gly Arg Met Val Pro Val 980 985 54349DNAHomo
sapiensCDS(445)..(3093) 5tttttgaatc ggttgtggcg gccgcggcga
ggaatggcgg tatttgtgag aggagtcggc 60gtttgaagag gtggaactcc tagggctttt
ttgagagtga cggagtctac ctcttgttac 120ctagactgga gtgcagtggc
acgatctcgg ctcactgcaa cctctgcctc ccgggttcaa 180gcgattctcc
tgcctcagcc tcctgagtag ctgggattac aggtgcctgc caccaagccc
240agctaatttt tgtattttta gtagagatgg ggtttcattg tgttggccag
gctggtctcg 300aactcctgac ctcgtgatcc gcccgccttg gcctcccaaa
gtgctaggat tacaagtgtg 360agccaccgcg tccggccttt caaatggtat
ttttgatttt cctcttccag tccttaaagc 420agctgattta gaagaataca aatc atg
gct gaa aat agt gta tta aca tcc 471 Met Ala Glu Asn Ser Val Leu Thr
Ser 1 5 act act ggg agg act agc ttg gca gac tct tcc att ttt gat tct
aaa 519Thr Thr Gly Arg Thr Ser Leu Ala Asp Ser Ser Ile Phe Asp Ser
Lys 10 15 20 25 gtt act gag att tcc aag gaa aac tta ctt att gga tct
act tca tat 567Val Thr Glu Ile Ser Lys Glu Asn Leu Leu Ile Gly Ser
Thr Ser Tyr 30 35 40 gta gaa gag atg cct cag att gaa aca aga gtg
ata ttg gtt caa gaa 615Val Glu Glu Met Pro Gln Ile Glu Thr Arg Val
Ile Leu Val Gln Glu 45 50 55 gct gga aaa caa gaa gaa ctt ata aaa
gcc tta aag gac att aaa gtg 663Ala Gly Lys Gln Glu Glu Leu Ile Lys
Ala Leu Lys Asp Ile Lys Val 60 65 70 ggc ttt gta aag atg gag tca
gtg gaa gaa ttt gaa ggt ttg gat tct 711Gly Phe Val Lys Met Glu Ser
Val Glu Glu Phe Glu Gly Leu Asp Ser 75 80 85 ccg gaa ttt gaa aat
gta ttt gta gtc acg gac ttt cag gat tct gtc 759Pro Glu Phe Glu Asn
Val Phe Val Val Thr Asp Phe Gln Asp Ser Val 90 95 100 105 ttt aat
gac ctc tac aag gct gat tgt aga gtt att gga cca cca gtt 807Phe Asn
Asp Leu Tyr Lys Ala Asp Cys Arg Val Ile Gly Pro Pro Val 110 115 120
gta tta aat tgt tca caa aaa gga gag cct ttg cca ttt tca tgt cgc
855Val Leu Asn Cys Ser Gln Lys Gly Glu Pro Leu Pro Phe Ser Cys Arg
125 130 135 ccg ttg tat tgt aca agt atg atg aat cta gta cta tgc ttt
act gga 903Pro Leu Tyr Cys Thr Ser Met Met Asn Leu Val Leu Cys Phe
Thr Gly 140 145 150 ttt agg aaa aaa gaa gaa cta gtc agg ttg gtg aca
ttg gtc cat cac 951Phe Arg Lys Lys Glu Glu Leu Val Arg Leu Val Thr
Leu Val His His 155 160 165 atg ggt gga gtt att cga aaa gac ttt aat
tca aaa gtt aca cat ttg 999Met Gly Gly Val Ile Arg Lys Asp Phe Asn
Ser Lys Val Thr His Leu 170 175 180 185 gtg gca aat tgt aca caa gga
gaa aaa ttc agg gtt gct gtg agt cta 1047Val Ala Asn Cys Thr Gln Gly
Glu Lys Phe Arg Val Ala Val Ser Leu 190 195 200 ggt act cca att atg
aag cca gaa tgg att tat aaa gct tgg gaa agg 1095Gly Thr Pro Ile Met
Lys Pro Glu Trp Ile Tyr Lys Ala Trp Glu Arg 205 210 215 cgg aat gaa
cag gat ttc tat gca gca gtt gat gac ttt aga aat gaa 1143Arg Asn Glu
Gln Asp Phe Tyr Ala Ala Val Asp Asp Phe Arg Asn Glu 220 225 230 ttt
aaa gtt cct cca ttt caa gat tgt att tta agt ttc ctg gga ttt 1191Phe
Lys Val Pro Pro Phe Gln Asp Cys Ile Leu Ser Phe Leu Gly Phe 235 240
245 tca gat gaa gag aaa acc aat atg gaa gaa atg act gaa atg caa gga
1239Ser Asp Glu Glu Lys Thr Asn Met Glu Glu Met Thr Glu Met Gln Gly
250 255 260 265 ggt aaa tat tta ccg ctt gga gat gaa aga tgc act cac
ctt gta gtt 1287Gly Lys Tyr Leu Pro Leu Gly Asp Glu Arg Cys Thr His
Leu Val Val 270 275 280 gaa gag aat ata gta aaa gat ctt ccc ttt gaa
cct tca aag aaa ctt 1335Glu Glu Asn Ile Val Lys Asp Leu Pro Phe Glu
Pro Ser Lys Lys Leu 285 290 295 tat gtt gtc aag caa gag tgg ttc tgg
gga agc att caa atg gat gcc 1383Tyr Val Val Lys Gln Glu Trp Phe Trp
Gly Ser Ile Gln Met Asp Ala 300 305 310 cga gct gga gaa act atg tat
tta tat gaa aag gca aat act cct gag 1431Arg Ala Gly Glu Thr Met Tyr
Leu Tyr Glu Lys Ala Asn Thr Pro Glu 315 320 325 ctc aag aaa tca gtg
tca atg ctt tct cta aat acc cct aac agc aat 1479Leu Lys Lys Ser Val
Ser Met Leu Ser Leu Asn Thr Pro Asn Ser Asn 330 335 340 345 cgc aaa
cga cgt cgt tta aaa gaa aca ctt gct cag ctt tca aga gag 1527Arg Lys
Arg Arg Arg Leu Lys Glu Thr Leu Ala Gln Leu Ser Arg Glu 350 355 360
aca gac gtg tca cca ttt cca ccc cgt aag cgc cca tca gct gag cat
1575Thr Asp Val Ser Pro Phe Pro Pro Arg Lys Arg Pro Ser Ala Glu His
365 370 375 tcc ctt tcc ata ggg tca ctc cta gat atc tcc aac aca cca
gag tct 1623Ser Leu Ser Ile Gly Ser Leu Leu Asp Ile Ser Asn Thr Pro
Glu Ser 380 385 390 agc att aac tat gga gac acc cca aag tct tgt act
aag tct tct aaa 1671Ser Ile Asn Tyr Gly Asp Thr Pro Lys Ser Cys Thr
Lys Ser Ser Lys 395 400 405 agc tcc act cca gtt cct tca aag cag tca
gca agg tgg caa gtt gca 1719Ser Ser Thr Pro Val Pro Ser Lys Gln Ser
Ala Arg Trp Gln Val Ala 410 415 420 425 aaa gag ctt tat caa act gaa
agt aat tat gtt aat ata ttg gca aca 1767Lys Glu Leu Tyr Gln Thr Glu
Ser Asn Tyr Val Asn Ile Leu Ala Thr 430 435 440 att att cag tta ttt
caa gta cca ttg gaa gag gaa gga caa cgt ggt 1815Ile Ile Gln Leu Phe
Gln Val Pro Leu Glu Glu Glu Gly Gln Arg Gly 445 450 455 gga cct atc
ctt gca cca gag gag att aag act att ttt ggt agc atc 1863Gly Pro Ile
Leu Ala Pro Glu Glu Ile Lys Thr Ile Phe Gly Ser Ile 460 465 470 cca
gat atc ttt gat gta cac act aag ata aag gat gat ctt gaa gac 1911Pro
Asp Ile Phe Asp Val His Thr Lys Ile Lys Asp Asp Leu Glu Asp 475 480
485 ctt ata gtt aat tgg gat gag agc aaa agc att ggt gac att ttt ctg
1959Leu Ile Val Asn Trp Asp Glu Ser Lys Ser Ile Gly Asp Ile Phe Leu
490 495 500 505 aaa tat tca aaa gat ttg gta aaa acc tac cct ccc ttt
gta aac ttc 2007Lys Tyr Ser Lys Asp Leu Val Lys Thr Tyr Pro Pro Phe
Val Asn Phe 510 515 520 ttt gaa atg agc aag gaa aca att att aaa tgt
gaa aaa cag aaa cca 2055Phe Glu Met Ser Lys Glu Thr Ile Ile Lys Cys
Glu Lys Gln Lys Pro 525 530 535 aga ttt cat gct ttt ctc aag ata aac
caa gca aaa cca gaa tgt gga 2103Arg Phe His Ala Phe Leu Lys Ile Asn
Gln Ala Lys Pro Glu Cys Gly 540 545 550 cgg cag agc ctt gtt gaa ctt
ctt atc cga cca gta cag agg tta ccc 2151Arg Gln Ser Leu Val Glu Leu
Leu Ile Arg Pro Val Gln Arg Leu Pro 555 560 565 agt gtt gca tta ctt
tta aat gat ctt aag aag cat aca gct gat gaa 2199Ser Val Ala Leu Leu
Leu Asn Asp Leu Lys Lys His Thr Ala Asp Glu 570 575 580 585 aat cca
gac aaa agc act tta gaa aaa gct att gga tca ctg aag gaa 2247Asn Pro
Asp Lys Ser Thr Leu Glu Lys Ala Ile Gly Ser Leu Lys Glu 590 595 600
gta atg acg cat att aat gag gat aag aga aaa aca gaa gct caa aag
2295Val Met Thr His Ile Asn Glu Asp Lys Arg Lys Thr Glu Ala Gln Lys
605 610 615 caa att ttt gat gtt gtt tat gaa gta gat gga tgc cca gct
aat ctt 2343Gln Ile Phe Asp Val Val Tyr Glu Val Asp Gly Cys Pro Ala
Asn Leu 620 625 630 tta tct tct cac cga agc tta gta cag cgg gtt gaa
aca att tct cta 2391Leu Ser Ser His Arg Ser Leu Val Gln Arg Val Glu
Thr Ile Ser Leu 635 640 645 ggt gag cac ccc tgt gac aga gga gaa caa
gta act ctc ttc ctc ttc 2439Gly Glu His Pro Cys Asp Arg Gly Glu Gln
Val Thr Leu Phe Leu Phe 650 655 660 665 aat gat tgc cta gag ata gca
aga aaa cgg cac aag gtt att ggc act 2487Asn Asp Cys Leu Glu Ile Ala
Arg Lys Arg His Lys Val Ile Gly Thr 670 675 680 ttt agg agt cct cat
ggc caa acc cga ccc cca gct tct ctt aag cat 2535Phe Arg Ser Pro His
Gly Gln Thr Arg Pro Pro Ala Ser Leu Lys His 685 690 695 att cac cta
atg cct ctt tct cag att aag aag gta ttg gac ata aga 2583Ile His Leu
Met Pro Leu Ser Gln Ile Lys Lys Val Leu Asp Ile Arg 700 705 710 gag
aca gaa gat tgc cat aat gct ttt gcc ttg ctt gtg agg cca cca 2631Glu
Thr Glu Asp Cys His Asn Ala Phe Ala Leu Leu Val Arg Pro Pro 715 720
725 aca gag cag gca aat gtg cta ctc agt ttc cag atg aca tca gat gaa
2679Thr Glu Gln Ala Asn Val Leu Leu Ser Phe Gln Met Thr Ser Asp Glu
730 735 740 745 ctt cca aaa gaa aac tgg cta aag atg ctg tgt cga cat
gta gct aac 2727Leu Pro Lys Glu Asn Trp Leu Lys Met Leu Cys Arg His
Val Ala Asn 750 755 760 acc att tgt aaa gca gat gct gag aat ctt att
tat act gct gat cca 2775Thr Ile Cys Lys Ala Asp Ala Glu Asn Leu Ile
Tyr Thr Ala Asp Pro 765 770 775 gaa tcc ttt gaa gta aat aca aaa gat
atg gac agt aca ttg agt aga 2823Glu Ser Phe Glu Val Asn Thr Lys Asp
Met Asp Ser Thr Leu Ser Arg 780 785 790 gca tca aga gca ata aaa aag
act tca aaa aag gtt aca aga gca ttc 2871Ala Ser Arg Ala Ile Lys Lys
Thr Ser Lys Lys Val Thr Arg Ala Phe 795 800 805 tct ttc tcc aaa act
cca aaa aga gct ctt cga agg gct ctt atg aca 2919Ser Phe Ser Lys Thr
Pro Lys Arg Ala Leu Arg Arg Ala Leu Met Thr 810 815 820 825 tcc cac
ggc tca gtg gag gga aga agt cct tcc agc aat gat aag cat 2967Ser His
Gly Ser Val Glu Gly Arg Ser Pro Ser Ser Asn Asp Lys His 830 835 840
gta atg agt cgt ctt tct agc aca tca tca tta gca ggt atc cct tct
3015Val Met Ser Arg Leu Ser Ser Thr Ser Ser Leu Ala Gly Ile Pro Ser
845 850 855 ccc tcc ctt gtc agc ctt cct tcc ttc ttt gaa agg aga agt
cat acg 3063Pro Ser Leu Val Ser Leu Pro Ser Phe Phe Glu Arg Arg Ser
His Thr 860 865 870 tta agt aga tct aca act cat ttg ata tga
agcgttacca aaatcttaaa 3113Leu Ser Arg Ser Thr Thr His Leu Ile 875
880 ttatagaaat gtatagacac ctcatactca aataagaaac tgacttaaat
ggtacttgta 3173attagcacgt tggtgaaagc tggaaggaag ataaataaca
ctaaactatg ctatttgatt 3233tttcttcttg aaagagtaag gtttacctgt
tacattttca agttaattca tgtaaaaaat 3293gatagtgatt ttgatgtaat
ttatctcttg tttgaatctg tcattcaaag gccaataatt 3353taagttgcta
tcagctgata ttagtagctt tgcaaccctg atagagtaaa taaattttat
3413gggtgggtgc caaatactgc tgtgaatcta tttgtatagt atccatgaat
gaatttatgg 3473aaatagatat ttgtgcagct caatttatgc agagattaaa
tgacatcata atactggatg 3533aaaacttgca tagaattctg attaaatagt
gggtctgttt cacatgtgca gtttgaagta 3593tttaaataac cactcctttc
acagtttatt ttcttctcaa gcgttttcaa gatctagcat 3653gtggatttta
aaagatttgc cctcattaac aagaataaca tttaaaggag attgtttcaa
3713aatatttttg caaattgaga taaggacaga aagattgaga aacattgtat
attttgcaaa
3773aacaagatgt ttgtagctgt ttcagagaga gtacggtata tttatggtaa
ttttatccac 3833tagcaaatct tgatttagtt tgatagtcgt cgtcggaatt
ttattttgaa ggataagacc 3893atgggaaaat tgtggtaaag actgtttgta
cccttcatga aataattctg aagttgccat 3953cagttttact aatcttctgt
gaaatgcata gatatgcgca tgttcaactt tttattgtgg 4013tcttataatt
aaatgtaaaa ttgaaaattc atttgctgtt tcaaagtgtg atatctttca
4073caatagcctt tttatagtca gtaattcaga ataatcaagt tcatatggat
aaatgcattt 4133ttatttccta tttctttagg gagtgctaca aatgtttgtc
acttaaattt caagtttctg 4193ttttaatagt taactgacta tagattgttt
tctatgccat gtatgtgcca cttctgagag 4253tagtaaatga ctctttgcta
cattttaaaa gcaattgtat tagtaagaac tttgtaaata 4313aatacctaaa
acccaagtgt aaaaaaaaaa aaaaaa 43496882PRTHomo sapiens 6Met Ala Glu
Asn Ser Val Leu Thr Ser Thr Thr Gly Arg Thr Ser Leu 1 5 10 15 Ala
Asp Ser Ser Ile Phe Asp Ser Lys Val Thr Glu Ile Ser Lys Glu 20 25
30 Asn Leu Leu Ile Gly Ser Thr Ser Tyr Val Glu Glu Met Pro Gln Ile
35 40 45 Glu Thr Arg Val Ile Leu Val Gln Glu Ala Gly Lys Gln Glu
Glu Leu 50 55 60 Ile Lys Ala Leu Lys Asp Ile Lys Val Gly Phe Val
Lys Met Glu Ser 65 70 75 80 Val Glu Glu Phe Glu Gly Leu Asp Ser Pro
Glu Phe Glu Asn Val Phe 85 90 95 Val Val Thr Asp Phe Gln Asp Ser
Val Phe Asn Asp Leu Tyr Lys Ala 100 105 110 Asp Cys Arg Val Ile Gly
Pro Pro Val Val Leu Asn Cys Ser Gln Lys 115 120 125 Gly Glu Pro Leu
Pro Phe Ser Cys Arg Pro Leu Tyr Cys Thr Ser Met 130 135 140 Met Asn
Leu Val Leu Cys Phe Thr Gly Phe Arg Lys Lys Glu Glu Leu 145 150 155
160 Val Arg Leu Val Thr Leu Val His His Met Gly Gly Val Ile Arg Lys
165 170 175 Asp Phe Asn Ser Lys Val Thr His Leu Val Ala Asn Cys Thr
Gln Gly 180 185 190 Glu Lys Phe Arg Val Ala Val Ser Leu Gly Thr Pro
Ile Met Lys Pro 195 200 205 Glu Trp Ile Tyr Lys Ala Trp Glu Arg Arg
Asn Glu Gln Asp Phe Tyr 210 215 220 Ala Ala Val Asp Asp Phe Arg Asn
Glu Phe Lys Val Pro Pro Phe Gln 225 230 235 240 Asp Cys Ile Leu Ser
Phe Leu Gly Phe Ser Asp Glu Glu Lys Thr Asn 245 250 255 Met Glu Glu
Met Thr Glu Met Gln Gly Gly Lys Tyr Leu Pro Leu Gly 260 265 270 Asp
Glu Arg Cys Thr His Leu Val Val Glu Glu Asn Ile Val Lys Asp 275 280
285 Leu Pro Phe Glu Pro Ser Lys Lys Leu Tyr Val Val Lys Gln Glu Trp
290 295 300 Phe Trp Gly Ser Ile Gln Met Asp Ala Arg Ala Gly Glu Thr
Met Tyr 305 310 315 320 Leu Tyr Glu Lys Ala Asn Thr Pro Glu Leu Lys
Lys Ser Val Ser Met 325 330 335 Leu Ser Leu Asn Thr Pro Asn Ser Asn
Arg Lys Arg Arg Arg Leu Lys 340 345 350 Glu Thr Leu Ala Gln Leu Ser
Arg Glu Thr Asp Val Ser Pro Phe Pro 355 360 365 Pro Arg Lys Arg Pro
Ser Ala Glu His Ser Leu Ser Ile Gly Ser Leu 370 375 380 Leu Asp Ile
Ser Asn Thr Pro Glu Ser Ser Ile Asn Tyr Gly Asp Thr 385 390 395 400
Pro Lys Ser Cys Thr Lys Ser Ser Lys Ser Ser Thr Pro Val Pro Ser 405
410 415 Lys Gln Ser Ala Arg Trp Gln Val Ala Lys Glu Leu Tyr Gln Thr
Glu 420 425 430 Ser Asn Tyr Val Asn Ile Leu Ala Thr Ile Ile Gln Leu
Phe Gln Val 435 440 445 Pro Leu Glu Glu Glu Gly Gln Arg Gly Gly Pro
Ile Leu Ala Pro Glu 450 455 460 Glu Ile Lys Thr Ile Phe Gly Ser Ile
Pro Asp Ile Phe Asp Val His 465 470 475 480 Thr Lys Ile Lys Asp Asp
Leu Glu Asp Leu Ile Val Asn Trp Asp Glu 485 490 495 Ser Lys Ser Ile
Gly Asp Ile Phe Leu Lys Tyr Ser Lys Asp Leu Val 500 505 510 Lys Thr
Tyr Pro Pro Phe Val Asn Phe Phe Glu Met Ser Lys Glu Thr 515 520 525
Ile Ile Lys Cys Glu Lys Gln Lys Pro Arg Phe His Ala Phe Leu Lys 530
535 540 Ile Asn Gln Ala Lys Pro Glu Cys Gly Arg Gln Ser Leu Val Glu
Leu 545 550 555 560 Leu Ile Arg Pro Val Gln Arg Leu Pro Ser Val Ala
Leu Leu Leu Asn 565 570 575 Asp Leu Lys Lys His Thr Ala Asp Glu Asn
Pro Asp Lys Ser Thr Leu 580 585 590 Glu Lys Ala Ile Gly Ser Leu Lys
Glu Val Met Thr His Ile Asn Glu 595 600 605 Asp Lys Arg Lys Thr Glu
Ala Gln Lys Gln Ile Phe Asp Val Val Tyr 610 615 620 Glu Val Asp Gly
Cys Pro Ala Asn Leu Leu Ser Ser His Arg Ser Leu 625 630 635 640 Val
Gln Arg Val Glu Thr Ile Ser Leu Gly Glu His Pro Cys Asp Arg 645 650
655 Gly Glu Gln Val Thr Leu Phe Leu Phe Asn Asp Cys Leu Glu Ile Ala
660 665 670 Arg Lys Arg His Lys Val Ile Gly Thr Phe Arg Ser Pro His
Gly Gln 675 680 685 Thr Arg Pro Pro Ala Ser Leu Lys His Ile His Leu
Met Pro Leu Ser 690 695 700 Gln Ile Lys Lys Val Leu Asp Ile Arg Glu
Thr Glu Asp Cys His Asn 705 710 715 720 Ala Phe Ala Leu Leu Val Arg
Pro Pro Thr Glu Gln Ala Asn Val Leu 725 730 735 Leu Ser Phe Gln Met
Thr Ser Asp Glu Leu Pro Lys Glu Asn Trp Leu 740 745 750 Lys Met Leu
Cys Arg His Val Ala Asn Thr Ile Cys Lys Ala Asp Ala 755 760 765 Glu
Asn Leu Ile Tyr Thr Ala Asp Pro Glu Ser Phe Glu Val Asn Thr 770 775
780 Lys Asp Met Asp Ser Thr Leu Ser Arg Ala Ser Arg Ala Ile Lys Lys
785 790 795 800 Thr Ser Lys Lys Val Thr Arg Ala Phe Ser Phe Ser Lys
Thr Pro Lys 805 810 815 Arg Ala Leu Arg Arg Ala Leu Met Thr Ser His
Gly Ser Val Glu Gly 820 825 830 Arg Ser Pro Ser Ser Asn Asp Lys His
Val Met Ser Arg Leu Ser Ser 835 840 845 Thr Ser Ser Leu Ala Gly Ile
Pro Ser Pro Ser Leu Val Ser Leu Pro 850 855 860 Ser Phe Phe Glu Arg
Arg Ser His Thr Leu Ser Arg Ser Thr Thr His 865 870 875 880 Leu Ile
71372DNAHomo sapiensCDS(206)..(397) 7gcacgagggc gcttttgtct
ccggtgagtt ttgtggcggg aagcttctgc gctggtgctt 60agtaaccgac tttcctccgg
actcctgcac gacctgctcc tacagccggc gatccactcc 120cggctgttcc
cccggagggt ccagaggcct ttcagaagga gaaggcagct ctgtttctct
180gcagaggagt agggtccttt cagcc atg aag cat gtg ttg aac ctc tac ctg
232 Met Lys His Val Leu Asn Leu Tyr Leu 1 5 tta ggt gtg gta ctg acc
cta ctc tcc atc ttc gtt aga gtg atg gag 280Leu Gly Val Val Leu Thr
Leu Leu Ser Ile Phe Val Arg Val Met Glu 10 15 20 25 tcc cta gaa ggc
tta cta gag agc cca tcg cct ggg acc tcc tgg acc 328Ser Leu Glu Gly
Leu Leu Glu Ser Pro Ser Pro Gly Thr Ser Trp Thr 30 35 40 acc aga
agc caa cta gcc aac aca gag ccc acc aag ggc ctt cca gac 376Thr Arg
Ser Gln Leu Ala Asn Thr Glu Pro Thr Lys Gly Leu Pro Asp 45 50 55
cat cca tcc aga agc atg tga taagacctcc ttccatactg gccatatttt 427His
Pro Ser Arg Ser Met 60 ggaacactga cctagacatg tccagatggg agtcccattc
ctagcagaca agctgagcac 487cgttgtaacc agagaactat tactaggcct
tgaagaacct gtctaactgg atgctcattg 547cctgggcaag gcctgtttag
gccggttgcg gtggctcatg cctgtaatcc tagcactttg 607ggaggctgag
gtgggtggat cacctgaggt caggagttcg agaccagcct cgccaacatg
667gcgaaacccc atctctacta aaaatacaaa agttagctgg gtgtggtggc
agaggcctgt 727aatcccagtt ccttgggagg ctgaggcggg agaattgctt
gaacccgggg acggaggttg 787cagtgaaccg agatcgcact gctgtaccca
gcctgggcca cagtgcaaga ctccatctca 847aaaaaaaaaa gaaaagaaaa
agcctgttta atgcacaggt gtgagtggat tgcttatggc 907tatgagatag
gttgatctcg cccttacccc ggggtctggt gtatgctgtg ctttcctcag
967cagtatggct ctgacatctc ttagatgtcc caacttcagc tgttgggaga
tggtgatatt 1027ttcaacccta cttcctaaac atctgtctgg ggttccttta
gtcttgaatg tcttatgctc 1087aattatttgg tgttgagcct ctcttccaca
agagctcctc catgtttgga tagcagttga 1147agaggttgtg tgggtgggct
gttgggagtg aggatggagt gttcagtgcc catttctcat 1207tttacatttt
aaagtcgttc ctccaacata gtgtgtattg gtctgaaggg ggtggtggga
1267tgccaaagcc tgctcaagtt atggacattg tggccaccat gtggcttaaa
tgattttttc 1327taactaataa agtggaatat atatttcaaa aaaaaaaaaa aaaaa
1372863PRTHomo sapiens 8Met Lys His Val Leu Asn Leu Tyr Leu Leu Gly
Val Val Leu Thr Leu 1 5 10 15 Leu Ser Ile Phe Val Arg Val Met Glu
Ser Leu Glu Gly Leu Leu Glu 20 25 30 Ser Pro Ser Pro Gly Thr Ser
Trp Thr Thr Arg Ser Gln Leu Ala Asn 35 40 45 Thr Glu Pro Thr Lys
Gly Leu Pro Asp His Pro Ser Arg Ser Met 50 55 60 93516DNAHomo
sapiensCDS(767)..(1990) 9ggatcctgga gacaactttg ccgtgtgacg
cgccgggagg actgcagggc ccgcggccga 60gggctcggcg ccgcctgtga gcgggcccgc
gcggccggct ctcccgggca ccaagcttgc 120tccgcgccac tgcccgccgg
cccgcggcga ggacgacctg cccgtctccg ccgccggcgg 180cccttcctgg
cgcgaggcag tgagggcgag gcgctcaggt gcgagcgcgg ggccccgccg
240cagcgcccgc cgcagcgccg cgccaagccg cgcccggctc cgctccgggg
ggctccagcg 300ccttcgcttc cgtctcagcc aagttgcgtg gacccgctct
ttcgccacct tccccagccg 360ccggccgaac cgccgctccc actgacgctg
ctttcgcttc acccgaaccg gggctgcggg 420gcccccgacg cggaaaggat
ggggagaagg ctgcagatgc cgaggcgccc cgagacgccc 480gtgcggcagt
gacccgcgac ctccgccccg cccggcgcgc ccctcgggcc cccggggccc
540tcggcgcccc ttccctgccg cgcgggaacc cccgaggccc ggccggcccc
ctccccctgc 600gagccggcgg cagccctccc ggcgggcggg cgggcggagg
cccgggcggg cgcgggcgcg 660ggcgggggcg gggcggggcg gcgcgcccgg
agcccggagc ccggccctgc gctcggctcg 720actcggctcg cctcgcggcg
ggcgccctcg tcgccagcgg cgcacc atg gac ggg 775 Met Asp Gly 1 ctg ccc
ggt cgg gcg ctg ggg gcc gcc tgc ctt ctg ctg ctg gcg gcc 823Leu Pro
Gly Arg Ala Leu Gly Ala Ala Cys Leu Leu Leu Leu Ala Ala 5 10 15 ggc
tgg ctg ggg cct gag gcc tgg ggc tca ccc acg ccc ccg ccg acg 871Gly
Trp Leu Gly Pro Glu Ala Trp Gly Ser Pro Thr Pro Pro Pro Thr 20 25
30 35 cct gcc gcg ccg ccg cca ccc ccg cca ccc gga gcc ccg ggt ggc
tcg 919Pro Ala Ala Pro Pro Pro Pro Pro Pro Pro Gly Ala Pro Gly Gly
Ser 40 45 50 cag gac acc tgt acg tcg tgc ggc ggc ttc cgg cgg cca
gag gag ctc 967Gln Asp Thr Cys Thr Ser Cys Gly Gly Phe Arg Arg Pro
Glu Glu Leu 55 60 65 ggc cga gtg gac ggc gac ttc ctg gag gcg gtg
aag cgg cac atc ttg 1015Gly Arg Val Asp Gly Asp Phe Leu Glu Ala Val
Lys Arg His Ile Leu 70 75 80 agc cgc ctg cag atg cgg ggc cgg ccc
aac atc acg cac gcc gtg cct 1063Ser Arg Leu Gln Met Arg Gly Arg Pro
Asn Ile Thr His Ala Val Pro 85 90 95 aag gcc gcc atg gtc acg gcc
ctg cgc aag ctg cac gcg ggc aag gtg 1111Lys Ala Ala Met Val Thr Ala
Leu Arg Lys Leu His Ala Gly Lys Val 100 105 110 115 cgc gag gac ggc
cgc gtg gag atc ccg cac ctc gac ggc cac gcc agc 1159Arg Glu Asp Gly
Arg Val Glu Ile Pro His Leu Asp Gly His Ala Ser 120 125 130 ccg ggc
gcc gac ggc cag gag cgc gtt tcc gaa atc atc agc ttc gcc 1207Pro Gly
Ala Asp Gly Gln Glu Arg Val Ser Glu Ile Ile Ser Phe Ala 135 140 145
gag aca gat ggc ctc gcc tcc tcc cgg gtc cgc cta tac ttc ttc atc
1255Glu Thr Asp Gly Leu Ala Ser Ser Arg Val Arg Leu Tyr Phe Phe Ile
150 155 160 tcc aac gaa ggc aac cag aac ctg ttt gtg gtc cag gcc agc
ctg tgg 1303Ser Asn Glu Gly Asn Gln Asn Leu Phe Val Val Gln Ala Ser
Leu Trp 165 170 175 ctt tac ctg aaa ctc ctg ccc tac gtc ctg gag aag
ggc agc cgg cgg 1351Leu Tyr Leu Lys Leu Leu Pro Tyr Val Leu Glu Lys
Gly Ser Arg Arg 180 185 190 195 aag gtg cgg gtc aaa gtg tac ttc cag
gag cag ggc cac ggt gac agg 1399Lys Val Arg Val Lys Val Tyr Phe Gln
Glu Gln Gly His Gly Asp Arg 200 205 210 tgg aac atg gtg gag aag agg
gtg gac ctc aag cgc agc ggc tgg cat 1447Trp Asn Met Val Glu Lys Arg
Val Asp Leu Lys Arg Ser Gly Trp His 215 220 225 acc ttc cca ctc acg
gag gcc atc cag gcc ttg ttt gag cgg ggc gag 1495Thr Phe Pro Leu Thr
Glu Ala Ile Gln Ala Leu Phe Glu Arg Gly Glu 230 235 240 cgg cga ctc
aac cta gac gtg cag tgt gac agc tgc cag gag ctg gcc 1543Arg Arg Leu
Asn Leu Asp Val Gln Cys Asp Ser Cys Gln Glu Leu Ala 245 250 255 gtg
gtg ccg gtg ttc gtg gac cca ggc gaa gag tcg cac cga ccc ttt 1591Val
Val Pro Val Phe Val Asp Pro Gly Glu Glu Ser His Arg Pro Phe 260 265
270 275 gtg gtg gtg cag gct cgg ctg ggc gac agc agg cac cgc att cgc
aag 1639Val Val Val Gln Ala Arg Leu Gly Asp Ser Arg His Arg Ile Arg
Lys 280 285 290 cga ggc ctg gag tgc gat ggc cgg acc aac ctc tgt tgc
agg caa cag 1687Arg Gly Leu Glu Cys Asp Gly Arg Thr Asn Leu Cys Cys
Arg Gln Gln 295 300 305 ttc ttc att gac ttc cgc ctc atc ggc tgg aac
gac tgg atc ata gca 1735Phe Phe Ile Asp Phe Arg Leu Ile Gly Trp Asn
Asp Trp Ile Ile Ala 310 315 320 ccc acc ggc tac tac ggc aac tac tgt
gag ggc agc tgc cca gcc tac 1783Pro Thr Gly Tyr Tyr Gly Asn Tyr Cys
Glu Gly Ser Cys Pro Ala Tyr 325 330 335 ctg gca ggg gtc ccc ggc tct
gcc tcc tcc ttc cac acg gct gtg gtg 1831Leu Ala Gly Val Pro Gly Ser
Ala Ser Ser Phe His Thr Ala Val Val 340 345 350 355 aac cag tac cgc
atg cgg ggt ctg aac ccc ggc acg gtg aac tcc tgc 1879Asn Gln Tyr Arg
Met Arg Gly Leu Asn Pro Gly Thr Val Asn Ser Cys 360 365 370 tgc att
ccc acc aag ctg agc acc atg tcc atg ctg tac ttc gat gat 1927Cys Ile
Pro Thr Lys Leu Ser Thr Met Ser Met Leu Tyr Phe Asp Asp 375 380 385
gag tac aac atc gtc aag cgg gac gtg ccc aac atg att gtg gag gag
1975Glu Tyr Asn Ile Val Lys Arg Asp Val Pro Asn Met Ile Val Glu Glu
390 395 400 tgc ggc tgc gcc tga cagtgcaagg caggggcacg gtggtggggc
acggagggca 2030Cys Gly Cys Ala 405 gtcccgggtg ggcttcttcc agccccccgc
gggaacgggg tacacggtgg gctgagtaca 2090gtcattctgt tgggctgtgg
agatagtgcc agggtgcggc ctgagatatt tttctacagc 2150ttcatagagc
aaccagtcaa aaccagagcg agaaccctca actgacatga aatactttaa
2210aatgcacacg tagccacgca cagccagacg catcctgcca cccacacagc
agcctccagg 2270ataccagcaa atggatgcgg tgacaaatgg cagcttagct
acaaatgcct gtcagtcgga 2330gagaatgggg tgagcagcca ccattccacc
agctggcccg gccacgtctc gaagttgcgc 2390cttcccgagc acacataaaa
gcacaaagac agagacgcag agagagagag agagccacgg 2450agaggaaaag
cagatgcagg ggtggggagc gcagctcggc ggaggctgcg tgtgccccgt
2510ggcttttacc aggcctgctc tgcctggctc gatgtctgct tcttcccagc
ctgggatcct 2570tcgtgcttca aggcctgggg agcctgtcct tccatgccct
tgtcgaggga aagagaccca
2630gaaaggacac aacccgtcag agacctggga gcaggggcaa tgaccgtttg
actgtttgtg 2690gcttgggcct ctgacatgac ttatgtgtgt gtgtgttttt
ggggtgggga gggagggaga 2750gaagaggggg ctaaatttga tgctttaact
gatctccaac agttgacagg tcatccttgc 2810cagttgtata actgaaaaag
gacttttcta ccaggtatga ccttttaagt gaaaatctga 2870attgttctaa
atggaaagaa aaaaagttgc aatctgtgcc cttcattggg gacattcctc
2930taggactggt ttggggacgg gtgggaatga cccctaggca aggggatgag
accgcaggag 2990gaaatggcgg ggaggtggca ttcttgaact gctgaggatg
gggggtgtcc cctcagcgga 3050ggccaaggga ggggagcagc ctagttggtc
ttggagagat ggggaaggct ttcagctgat 3110ttgcagaagt tgcccatgtg
ggcccaacca tcagggctgg ccgtggacgt ggcccctgcc 3170cactcacctg
cccgcctgcc cgcccgcccg catagcactt gcagacctgc ctgaacgcac
3230atgacatagc acttgccgat ctgcgtgtgc ccagaagtgg cccttggccg
agcgccgaac 3290tcgctcgccc tctagatgtc caagtgccac gtgaactatg
caatttaaag ggttgaccca 3350cactagacga aactggactc gtacgactct
ttttatattt tttatacttg aaatgaaatc 3410ctttgcttct tttttaagcg
aatgattgct tttaatgttt gcactgattt agttgcatga 3470ttagtcagaa
actgccattt gaaaaaaaag ttatttttat agcagc 351610407PRTHomo sapiens
10Met Asp Gly Leu Pro Gly Arg Ala Leu Gly Ala Ala Cys Leu Leu Leu 1
5 10 15 Leu Ala Ala Gly Trp Leu Gly Pro Glu Ala Trp Gly Ser Pro Thr
Pro 20 25 30 Pro Pro Thr Pro Ala Ala Pro Pro Pro Pro Pro Pro Pro
Gly Ala Pro 35 40 45 Gly Gly Ser Gln Asp Thr Cys Thr Ser Cys Gly
Gly Phe Arg Arg Pro 50 55 60 Glu Glu Leu Gly Arg Val Asp Gly Asp
Phe Leu Glu Ala Val Lys Arg 65 70 75 80 His Ile Leu Ser Arg Leu Gln
Met Arg Gly Arg Pro Asn Ile Thr His 85 90 95 Ala Val Pro Lys Ala
Ala Met Val Thr Ala Leu Arg Lys Leu His Ala 100 105 110 Gly Lys Val
Arg Glu Asp Gly Arg Val Glu Ile Pro His Leu Asp Gly 115 120 125 His
Ala Ser Pro Gly Ala Asp Gly Gln Glu Arg Val Ser Glu Ile Ile 130 135
140 Ser Phe Ala Glu Thr Asp Gly Leu Ala Ser Ser Arg Val Arg Leu Tyr
145 150 155 160 Phe Phe Ile Ser Asn Glu Gly Asn Gln Asn Leu Phe Val
Val Gln Ala 165 170 175 Ser Leu Trp Leu Tyr Leu Lys Leu Leu Pro Tyr
Val Leu Glu Lys Gly 180 185 190 Ser Arg Arg Lys Val Arg Val Lys Val
Tyr Phe Gln Glu Gln Gly His 195 200 205 Gly Asp Arg Trp Asn Met Val
Glu Lys Arg Val Asp Leu Lys Arg Ser 210 215 220 Gly Trp His Thr Phe
Pro Leu Thr Glu Ala Ile Gln Ala Leu Phe Glu 225 230 235 240 Arg Gly
Glu Arg Arg Leu Asn Leu Asp Val Gln Cys Asp Ser Cys Gln 245 250 255
Glu Leu Ala Val Val Pro Val Phe Val Asp Pro Gly Glu Glu Ser His 260
265 270 Arg Pro Phe Val Val Val Gln Ala Arg Leu Gly Asp Ser Arg His
Arg 275 280 285 Ile Arg Lys Arg Gly Leu Glu Cys Asp Gly Arg Thr Asn
Leu Cys Cys 290 295 300 Arg Gln Gln Phe Phe Ile Asp Phe Arg Leu Ile
Gly Trp Asn Asp Trp 305 310 315 320 Ile Ile Ala Pro Thr Gly Tyr Tyr
Gly Asn Tyr Cys Glu Gly Ser Cys 325 330 335 Pro Ala Tyr Leu Ala Gly
Val Pro Gly Ser Ala Ser Ser Phe His Thr 340 345 350 Ala Val Val Asn
Gln Tyr Arg Met Arg Gly Leu Asn Pro Gly Thr Val 355 360 365 Asn Ser
Cys Cys Ile Pro Thr Lys Leu Ser Thr Met Ser Met Leu Tyr 370 375 380
Phe Asp Asp Glu Tyr Asn Ile Val Lys Arg Asp Val Pro Asn Met Ile 385
390 395 400 Val Glu Glu Cys Gly Cys Ala 405 112972DNAHomo
sapiensCDS(28)..(2700) 11tcttcggacc taggctgccc tgccgtc atg tcg caa
ggg atc ctt tct ccg cca 54 Met Ser Gln Gly Ile Leu Ser Pro Pro 1 5
gcg ggc ttg ctg tcc gat gac gat gtc gta gtt tct ccc atg ttt gag
102Ala Gly Leu Leu Ser Asp Asp Asp Val Val Val Ser Pro Met Phe Glu
10 15 20 25 tcc aca gct gca gat ttg ggg tct gtg gta cgc aag aac ctg
cta tca 150Ser Thr Ala Ala Asp Leu Gly Ser Val Val Arg Lys Asn Leu
Leu Ser 30 35 40 gac tgc tct gtc gtc tct acc tcc cta gag gac aag
cag cag gtt cca 198Asp Cys Ser Val Val Ser Thr Ser Leu Glu Asp Lys
Gln Gln Val Pro 45 50 55 tct gag gac agt atg gag aag gtg aaa gta
tac ttg agg gtt agg ccc 246Ser Glu Asp Ser Met Glu Lys Val Lys Val
Tyr Leu Arg Val Arg Pro 60 65 70 ttg tta cct tca gag ttg gaa cga
cag gaa gat cag ggt tgt gtc cgt 294Leu Leu Pro Ser Glu Leu Glu Arg
Gln Glu Asp Gln Gly Cys Val Arg 75 80 85 att gag aat gtg gag acc
ctt gtt cta caa gca ccc aag gac tcg ttt 342Ile Glu Asn Val Glu Thr
Leu Val Leu Gln Ala Pro Lys Asp Ser Phe 90 95 100 105 gcc ctg aag
agc aat gaa cgg gga att ggc caa gcc aca cac agg ttc 390Ala Leu Lys
Ser Asn Glu Arg Gly Ile Gly Gln Ala Thr His Arg Phe 110 115 120 acc
ttt tcc cag atc ttt ggg cca gaa gtg gga cag gca tcc ttc ttc 438Thr
Phe Ser Gln Ile Phe Gly Pro Glu Val Gly Gln Ala Ser Phe Phe 125 130
135 aac cta act gtg aag gag atg gta aag gat gta ctc aaa ggg cag aac
486Asn Leu Thr Val Lys Glu Met Val Lys Asp Val Leu Lys Gly Gln Asn
140 145 150 tgg ctc atc tat aca tat gga gtc act aac tca ggg aaa acc
cac acg 534Trp Leu Ile Tyr Thr Tyr Gly Val Thr Asn Ser Gly Lys Thr
His Thr 155 160 165 att caa ggt acc atc aag gat gga ggg att ctc ccc
cgg tcc ctg gcg 582Ile Gln Gly Thr Ile Lys Asp Gly Gly Ile Leu Pro
Arg Ser Leu Ala 170 175 180 185 ctg atc ttc aat agc ctc caa ggc caa
ctt cat cca aca cct gat ctg 630Leu Ile Phe Asn Ser Leu Gln Gly Gln
Leu His Pro Thr Pro Asp Leu 190 195 200 aag ccc ttg ctc tcc aat gag
gta atc tgg cta gac agc aag cag atc 678Lys Pro Leu Leu Ser Asn Glu
Val Ile Trp Leu Asp Ser Lys Gln Ile 205 210 215 cga cag gag gaa atg
aag aag ctg tcc ctg cta aat gga ggc ctc caa 726Arg Gln Glu Glu Met
Lys Lys Leu Ser Leu Leu Asn Gly Gly Leu Gln 220 225 230 gag gag gag
ctg tcc act tcc ttg aag agg agt gtc tac atc gaa agt 774Glu Glu Glu
Leu Ser Thr Ser Leu Lys Arg Ser Val Tyr Ile Glu Ser 235 240 245 cgg
ata ggt acc agc acc agc ttc gac agt ggc att gct ggg ctc tct 822Arg
Ile Gly Thr Ser Thr Ser Phe Asp Ser Gly Ile Ala Gly Leu Ser 250 255
260 265 tct atc agt cag tgt acc agc agt agc cag ctg gat gaa aca agt
cat 870Ser Ile Ser Gln Cys Thr Ser Ser Ser Gln Leu Asp Glu Thr Ser
His 270 275 280 cga tgg gca cag cca gac act gcc cca cta cct gtc ccg
gca aac att 918Arg Trp Ala Gln Pro Asp Thr Ala Pro Leu Pro Val Pro
Ala Asn Ile 285 290 295 cgc ttc tcc atc tgg atc tca ttc ttt gag atc
tac aac gaa ctg ctt 966Arg Phe Ser Ile Trp Ile Ser Phe Phe Glu Ile
Tyr Asn Glu Leu Leu 300 305 310 tat gac cta tta gaa ccg cct agc caa
cag cgc aag agg cag act ttg 1014Tyr Asp Leu Leu Glu Pro Pro Ser Gln
Gln Arg Lys Arg Gln Thr Leu 315 320 325 cgg cta tgc gag gat caa aat
ggc aat ccc tat gtg aaa gat ctc aac 1062Arg Leu Cys Glu Asp Gln Asn
Gly Asn Pro Tyr Val Lys Asp Leu Asn 330 335 340 345 tgg att cat gtg
caa gat gct gag gag gcc tgg aag ctc cta aaa gtg 1110Trp Ile His Val
Gln Asp Ala Glu Glu Ala Trp Lys Leu Leu Lys Val 350 355 360 ggt cgt
aag aac cag agc ttt gcc agc acc cac ctc aac cag aac tcc 1158Gly Arg
Lys Asn Gln Ser Phe Ala Ser Thr His Leu Asn Gln Asn Ser 365 370 375
agc cgc agt cac agc atc ttc tca atc agg atc cta cac ctt cag ggg
1206Ser Arg Ser His Ser Ile Phe Ser Ile Arg Ile Leu His Leu Gln Gly
380 385 390 gaa gga gat ata gtc ccc aag atc agc gag ctg tca ctc tgt
gat ctg 1254Glu Gly Asp Ile Val Pro Lys Ile Ser Glu Leu Ser Leu Cys
Asp Leu 395 400 405 gct ggc tca gag cgc tgc aaa gat cag aag agt ggt
gaa cgg ttg aag 1302Ala Gly Ser Glu Arg Cys Lys Asp Gln Lys Ser Gly
Glu Arg Leu Lys 410 415 420 425 gaa gca gga aac att aac acc tct cta
cac acc ctg ggc cgc tgt att 1350Glu Ala Gly Asn Ile Asn Thr Ser Leu
His Thr Leu Gly Arg Cys Ile 430 435 440 gct gcc ctt cgt caa aac cag
cag aac cgg tca aag cag aac ctg gtt 1398Ala Ala Leu Arg Gln Asn Gln
Gln Asn Arg Ser Lys Gln Asn Leu Val 445 450 455 ccc ttc cgt gac agc
aag ttg act cga gtg ttc caa ggt ttc ttc aca 1446Pro Phe Arg Asp Ser
Lys Leu Thr Arg Val Phe Gln Gly Phe Phe Thr 460 465 470 ggc cga ggc
cgt tcc tgc atg att gtc aat gtg aat ccc tgt gca tct 1494Gly Arg Gly
Arg Ser Cys Met Ile Val Asn Val Asn Pro Cys Ala Ser 475 480 485 acc
tat gat gaa act ctt cat gtg gcc aag ttc tca gcc att gct agc 1542Thr
Tyr Asp Glu Thr Leu His Val Ala Lys Phe Ser Ala Ile Ala Ser 490 495
500 505 cag ctt gtg cat gcc cca cct atg caa ctg gga ttc cca tcc ctg
cac 1590Gln Leu Val His Ala Pro Pro Met Gln Leu Gly Phe Pro Ser Leu
His 510 515 520 tcg ttc atc aag gaa cat agt ctt cag gta tcc ccc agc
tta gag aaa 1638Ser Phe Ile Lys Glu His Ser Leu Gln Val Ser Pro Ser
Leu Glu Lys 525 530 535 ggg gct aag gca gac aca ggc ctt gat gat gat
att gaa aat gaa gct 1686Gly Ala Lys Ala Asp Thr Gly Leu Asp Asp Asp
Ile Glu Asn Glu Ala 540 545 550 gac atc tcc atg tat ggc aaa gag gag
ctc cta caa gtt gtg gaa gcc 1734Asp Ile Ser Met Tyr Gly Lys Glu Glu
Leu Leu Gln Val Val Glu Ala 555 560 565 atg aag aca ctg ctt ttg aag
gaa cga cag gaa aag cta cag ctg gag 1782Met Lys Thr Leu Leu Leu Lys
Glu Arg Gln Glu Lys Leu Gln Leu Glu 570 575 580 585 atg cat ctc cga
gat gaa att tgc aat gag atg gta gaa cag atg caa 1830Met His Leu Arg
Asp Glu Ile Cys Asn Glu Met Val Glu Gln Met Gln 590 595 600 cag cgg
gaa cag tgg tgc agt gaa cat ttg gac acc caa aag gaa cta 1878Gln Arg
Glu Gln Trp Cys Ser Glu His Leu Asp Thr Gln Lys Glu Leu 605 610 615
ttg gag gaa atg tat gaa gaa aaa cta aat atc ctc aag gag tca ctg
1926Leu Glu Glu Met Tyr Glu Glu Lys Leu Asn Ile Leu Lys Glu Ser Leu
620 625 630 aca agt ttt tac caa gaa gag att cag gag cgg gat gaa aag
att gaa 1974Thr Ser Phe Tyr Gln Glu Glu Ile Gln Glu Arg Asp Glu Lys
Ile Glu 635 640 645 gag cta gaa gct ctc ttg cag gaa gcc aga caa cag
tca gtg gcc cat 2022Glu Leu Glu Ala Leu Leu Gln Glu Ala Arg Gln Gln
Ser Val Ala His 650 655 660 665 cag caa tca ggg tct gaa ttg gcc cta
cgg cgg tca caa agg ttg gca 2070Gln Gln Ser Gly Ser Glu Leu Ala Leu
Arg Arg Ser Gln Arg Leu Ala 670 675 680 gct tct gcc tcc acc cag cag
ctt cag gag gtt aaa gct aaa tta cag 2118Ala Ser Ala Ser Thr Gln Gln
Leu Gln Glu Val Lys Ala Lys Leu Gln 685 690 695 cag tgc aaa gca gag
cta aac tct acc act gaa gag ttg cat aag tat 2166Gln Cys Lys Ala Glu
Leu Asn Ser Thr Thr Glu Glu Leu His Lys Tyr 700 705 710 cag aaa atg
tta gaa cca cca ccc tca gcc aag ccc ttc acc att gat 2214Gln Lys Met
Leu Glu Pro Pro Pro Ser Ala Lys Pro Phe Thr Ile Asp 715 720 725 gtg
gac aag aag tta gaa gag ggc cag aag aat ata agg ctg ttg cgg 2262Val
Asp Lys Lys Leu Glu Glu Gly Gln Lys Asn Ile Arg Leu Leu Arg 730 735
740 745 aca gag ctt cag aaa ctt ggt gag tct ctc caa tca gca gag aga
gct 2310Thr Glu Leu Gln Lys Leu Gly Glu Ser Leu Gln Ser Ala Glu Arg
Ala 750 755 760 tgt tgc cac agc act ggg gca gga aaa ctt cgt caa gcc
ttg acc act 2358Cys Cys His Ser Thr Gly Ala Gly Lys Leu Arg Gln Ala
Leu Thr Thr 765 770 775 tgt gat gac atc tta atc aaa cag gac cag act
ctg gct gaa ctg cag 2406Cys Asp Asp Ile Leu Ile Lys Gln Asp Gln Thr
Leu Ala Glu Leu Gln 780 785 790 aac aac atg gtg cta gtg aaa ctg gac
ctt cgg aag aag gca gca tgt 2454Asn Asn Met Val Leu Val Lys Leu Asp
Leu Arg Lys Lys Ala Ala Cys 795 800 805 att gct gag cag tat cat act
gtg ttg aaa ctc caa ggc cag gtt tct 2502Ile Ala Glu Gln Tyr His Thr
Val Leu Lys Leu Gln Gly Gln Val Ser 810 815 820 825 gcc aaa aag cgc
ctt ggt acc aac cag gaa aat cag caa cca aac caa 2550Ala Lys Lys Arg
Leu Gly Thr Asn Gln Glu Asn Gln Gln Pro Asn Gln 830 835 840 caa cca
cca ggg aag aaa cca ttc ctt cga aat tta ctt ccc cga aca 2598Gln Pro
Pro Gly Lys Lys Pro Phe Leu Arg Asn Leu Leu Pro Arg Thr 845 850 855
cca acc tgc caa agc tca aca gac tgc agc cct tat gcc cgg atc cta
2646Pro Thr Cys Gln Ser Ser Thr Asp Cys Ser Pro Tyr Ala Arg Ile Leu
860 865 870 cgc tca cgg cgt tcc cct tta ctc aaa tct ggg cct ttt ggc
aaa aag 2694Arg Ser Arg Arg Ser Pro Leu Leu Lys Ser Gly Pro Phe Gly
Lys Lys 875 880 885 tac taa ggctgtgggg aaagagaaga gcagtcatgg
ccctgaggtg ggtcagctac 2750Tyr 890 tctcctgaag aaataggtct cttttatgct
ttaccatata tcaggaatta tatccaggat 2810gcaatactca gacactagct
tttttctcac ttttgtatta taaccaccta tgtaatctca 2870tgttgttgtt
tttttttatt tacttatatg atttctatgc acacaaaaac agttatatta
2930aagatattat tgttcacatt ttttattgaa aaaaaaaaaa aa 297212890PRTHomo
sapiens 12Met Ser Gln Gly Ile Leu Ser Pro Pro Ala Gly Leu Leu Ser
Asp Asp 1 5 10 15 Asp Val Val Val Ser Pro Met Phe Glu Ser Thr Ala
Ala Asp Leu Gly 20 25 30 Ser Val Val Arg Lys Asn Leu Leu Ser Asp
Cys Ser Val Val Ser Thr 35 40 45 Ser Leu Glu Asp Lys Gln Gln Val
Pro Ser Glu Asp Ser Met Glu Lys 50 55 60 Val Lys Val Tyr Leu Arg
Val Arg Pro Leu Leu Pro Ser Glu Leu Glu 65 70 75 80 Arg Gln Glu Asp
Gln Gly Cys Val Arg Ile Glu Asn Val Glu Thr Leu 85 90 95 Val Leu
Gln Ala Pro Lys Asp Ser Phe Ala Leu Lys Ser Asn Glu Arg 100 105 110
Gly Ile Gly Gln Ala Thr His Arg Phe Thr Phe Ser Gln Ile Phe Gly 115
120 125 Pro Glu Val Gly Gln Ala Ser Phe Phe Asn Leu Thr Val Lys Glu
Met 130 135 140
Val Lys Asp Val Leu Lys Gly Gln Asn Trp Leu Ile Tyr Thr Tyr Gly 145
150 155 160 Val Thr Asn Ser Gly Lys Thr His Thr Ile Gln Gly Thr Ile
Lys Asp 165 170 175 Gly Gly Ile Leu Pro Arg Ser Leu Ala Leu Ile Phe
Asn Ser Leu Gln 180 185 190 Gly Gln Leu His Pro Thr Pro Asp Leu Lys
Pro Leu Leu Ser Asn Glu 195 200 205 Val Ile Trp Leu Asp Ser Lys Gln
Ile Arg Gln Glu Glu Met Lys Lys 210 215 220 Leu Ser Leu Leu Asn Gly
Gly Leu Gln Glu Glu Glu Leu Ser Thr Ser 225 230 235 240 Leu Lys Arg
Ser Val Tyr Ile Glu Ser Arg Ile Gly Thr Ser Thr Ser 245 250 255 Phe
Asp Ser Gly Ile Ala Gly Leu Ser Ser Ile Ser Gln Cys Thr Ser 260 265
270 Ser Ser Gln Leu Asp Glu Thr Ser His Arg Trp Ala Gln Pro Asp Thr
275 280 285 Ala Pro Leu Pro Val Pro Ala Asn Ile Arg Phe Ser Ile Trp
Ile Ser 290 295 300 Phe Phe Glu Ile Tyr Asn Glu Leu Leu Tyr Asp Leu
Leu Glu Pro Pro 305 310 315 320 Ser Gln Gln Arg Lys Arg Gln Thr Leu
Arg Leu Cys Glu Asp Gln Asn 325 330 335 Gly Asn Pro Tyr Val Lys Asp
Leu Asn Trp Ile His Val Gln Asp Ala 340 345 350 Glu Glu Ala Trp Lys
Leu Leu Lys Val Gly Arg Lys Asn Gln Ser Phe 355 360 365 Ala Ser Thr
His Leu Asn Gln Asn Ser Ser Arg Ser His Ser Ile Phe 370 375 380 Ser
Ile Arg Ile Leu His Leu Gln Gly Glu Gly Asp Ile Val Pro Lys 385 390
395 400 Ile Ser Glu Leu Ser Leu Cys Asp Leu Ala Gly Ser Glu Arg Cys
Lys 405 410 415 Asp Gln Lys Ser Gly Glu Arg Leu Lys Glu Ala Gly Asn
Ile Asn Thr 420 425 430 Ser Leu His Thr Leu Gly Arg Cys Ile Ala Ala
Leu Arg Gln Asn Gln 435 440 445 Gln Asn Arg Ser Lys Gln Asn Leu Val
Pro Phe Arg Asp Ser Lys Leu 450 455 460 Thr Arg Val Phe Gln Gly Phe
Phe Thr Gly Arg Gly Arg Ser Cys Met 465 470 475 480 Ile Val Asn Val
Asn Pro Cys Ala Ser Thr Tyr Asp Glu Thr Leu His 485 490 495 Val Ala
Lys Phe Ser Ala Ile Ala Ser Gln Leu Val His Ala Pro Pro 500 505 510
Met Gln Leu Gly Phe Pro Ser Leu His Ser Phe Ile Lys Glu His Ser 515
520 525 Leu Gln Val Ser Pro Ser Leu Glu Lys Gly Ala Lys Ala Asp Thr
Gly 530 535 540 Leu Asp Asp Asp Ile Glu Asn Glu Ala Asp Ile Ser Met
Tyr Gly Lys 545 550 555 560 Glu Glu Leu Leu Gln Val Val Glu Ala Met
Lys Thr Leu Leu Leu Lys 565 570 575 Glu Arg Gln Glu Lys Leu Gln Leu
Glu Met His Leu Arg Asp Glu Ile 580 585 590 Cys Asn Glu Met Val Glu
Gln Met Gln Gln Arg Glu Gln Trp Cys Ser 595 600 605 Glu His Leu Asp
Thr Gln Lys Glu Leu Leu Glu Glu Met Tyr Glu Glu 610 615 620 Lys Leu
Asn Ile Leu Lys Glu Ser Leu Thr Ser Phe Tyr Gln Glu Glu 625 630 635
640 Ile Gln Glu Arg Asp Glu Lys Ile Glu Glu Leu Glu Ala Leu Leu Gln
645 650 655 Glu Ala Arg Gln Gln Ser Val Ala His Gln Gln Ser Gly Ser
Glu Leu 660 665 670 Ala Leu Arg Arg Ser Gln Arg Leu Ala Ala Ser Ala
Ser Thr Gln Gln 675 680 685 Leu Gln Glu Val Lys Ala Lys Leu Gln Gln
Cys Lys Ala Glu Leu Asn 690 695 700 Ser Thr Thr Glu Glu Leu His Lys
Tyr Gln Lys Met Leu Glu Pro Pro 705 710 715 720 Pro Ser Ala Lys Pro
Phe Thr Ile Asp Val Asp Lys Lys Leu Glu Glu 725 730 735 Gly Gln Lys
Asn Ile Arg Leu Leu Arg Thr Glu Leu Gln Lys Leu Gly 740 745 750 Glu
Ser Leu Gln Ser Ala Glu Arg Ala Cys Cys His Ser Thr Gly Ala 755 760
765 Gly Lys Leu Arg Gln Ala Leu Thr Thr Cys Asp Asp Ile Leu Ile Lys
770 775 780 Gln Asp Gln Thr Leu Ala Glu Leu Gln Asn Asn Met Val Leu
Val Lys 785 790 795 800 Leu Asp Leu Arg Lys Lys Ala Ala Cys Ile Ala
Glu Gln Tyr His Thr 805 810 815 Val Leu Lys Leu Gln Gly Gln Val Ser
Ala Lys Lys Arg Leu Gly Thr 820 825 830 Asn Gln Glu Asn Gln Gln Pro
Asn Gln Gln Pro Pro Gly Lys Lys Pro 835 840 845 Phe Leu Arg Asn Leu
Leu Pro Arg Thr Pro Thr Cys Gln Ser Ser Thr 850 855 860 Asp Cys Ser
Pro Tyr Ala Arg Ile Leu Arg Ser Arg Arg Ser Pro Leu 865 870 875 880
Leu Lys Ser Gly Pro Phe Gly Lys Lys Tyr 885 890 132150DNAHomo
sapiensCDS(105)..(2033) 13ctcgagccac gaaggccccg ctgtcctgtc
tagcagatac ttgcacggtt tacagaaatt 60cggtccctgg gtcgtgtcag gaaactggaa
aaaaggtcat aagc atg aag cgc agt 116 Met Lys Arg Ser 1 tca gtt tcc
agc ggt ggt gct ggc cgc ctc tcc atg cag gag tta aga 164Ser Val Ser
Ser Gly Gly Ala Gly Arg Leu Ser Met Gln Glu Leu Arg 5 10 15 20 tcc
cag gat gta aat aaa caa ggc ctc tat acc cct caa acc aaa gag 212Ser
Gln Asp Val Asn Lys Gln Gly Leu Tyr Thr Pro Gln Thr Lys Glu 25 30
35 aaa cca acc ttt gga aag ttg agt ata aac aaa ccg aca tct gaa aga
260Lys Pro Thr Phe Gly Lys Leu Ser Ile Asn Lys Pro Thr Ser Glu Arg
40 45 50 aaa gtc tcg cta ttt ggc aaa aga act agt gga cat gga tcc
cgg aat 308Lys Val Ser Leu Phe Gly Lys Arg Thr Ser Gly His Gly Ser
Arg Asn 55 60 65 agt caa ctt ggt ata ttt tcc agt tct gag aaa atc
aag gac ccg aga 356Ser Gln Leu Gly Ile Phe Ser Ser Ser Glu Lys Ile
Lys Asp Pro Arg 70 75 80 cca ctt aat gac aaa gca ttc att cag cag
tgt att cga caa ctc tgt 404Pro Leu Asn Asp Lys Ala Phe Ile Gln Gln
Cys Ile Arg Gln Leu Cys 85 90 95 100 gag ttt ctt aca gaa aat ggt
tat gca cat aat gtg tcc atg aaa tct 452Glu Phe Leu Thr Glu Asn Gly
Tyr Ala His Asn Val Ser Met Lys Ser 105 110 115 cta caa gct ccc tct
gtt aaa gac ttc ctg aag atc ttc aca ttt ctt 500Leu Gln Ala Pro Ser
Val Lys Asp Phe Leu Lys Ile Phe Thr Phe Leu 120 125 130 tat ggc ttc
ctg tgc ccc tca tac gaa ctt cct gac aca aag ttt gaa 548Tyr Gly Phe
Leu Cys Pro Ser Tyr Glu Leu Pro Asp Thr Lys Phe Glu 135 140 145 gaa
gag gtt cca aga atc ttt aaa gac ctt ggg tat cct ttt gca cta 596Glu
Glu Val Pro Arg Ile Phe Lys Asp Leu Gly Tyr Pro Phe Ala Leu 150 155
160 tcc aaa agc tcc atg tac aca gtg ggg gct cct cat aca tgg cct cac
644Ser Lys Ser Ser Met Tyr Thr Val Gly Ala Pro His Thr Trp Pro His
165 170 175 180 att gtg gca gcc tta gtt tgg cta ata gac tgc atc aag
ata cat act 692Ile Val Ala Ala Leu Val Trp Leu Ile Asp Cys Ile Lys
Ile His Thr 185 190 195 gcc atg aaa gaa agc tca cct tta ttt gat gat
ggg cag cct tgg gga 740Ala Met Lys Glu Ser Ser Pro Leu Phe Asp Asp
Gly Gln Pro Trp Gly 200 205 210 gaa gaa act gaa gat gga att atg cat
aat aag ttg ttt ttg gac tac 788Glu Glu Thr Glu Asp Gly Ile Met His
Asn Lys Leu Phe Leu Asp Tyr 215 220 225 acc ata aaa tgc tat gag agt
ttt atg agt ggt gcc gac agc ttt gat 836Thr Ile Lys Cys Tyr Glu Ser
Phe Met Ser Gly Ala Asp Ser Phe Asp 230 235 240 gag atg aat gca gag
ctg cag tca aaa ctg aag gat tta ttt aat gtg 884Glu Met Asn Ala Glu
Leu Gln Ser Lys Leu Lys Asp Leu Phe Asn Val 245 250 255 260 gat gct
ttt aag ctg gaa tca tta gaa gca aaa aac aga gca ttg aat 932Asp Ala
Phe Lys Leu Glu Ser Leu Glu Ala Lys Asn Arg Ala Leu Asn 265 270 275
gaa cag att gca aga ttg gaa caa gaa aga gaa aaa gaa ccg aat cgt
980Glu Gln Ile Ala Arg Leu Glu Gln Glu Arg Glu Lys Glu Pro Asn Arg
280 285 290 cta gag tcg ttg aga aaa ctg aag gct tcc tta caa gga gat
gtt caa 1028Leu Glu Ser Leu Arg Lys Leu Lys Ala Ser Leu Gln Gly Asp
Val Gln 295 300 305 aag tat cag gca tac atg agc aat ttg gag tct cat
tca gcc att ctt 1076Lys Tyr Gln Ala Tyr Met Ser Asn Leu Glu Ser His
Ser Ala Ile Leu 310 315 320 gac cag aaa tta aat ggt ctc aat gag gaa
att gct aga gta gaa cta 1124Asp Gln Lys Leu Asn Gly Leu Asn Glu Glu
Ile Ala Arg Val Glu Leu 325 330 335 340 gaa tgt gaa aca ata aaa cag
gag aac act cga cta cag aat atc att 1172Glu Cys Glu Thr Ile Lys Gln
Glu Asn Thr Arg Leu Gln Asn Ile Ile 345 350 355 gac aac cag aag tac
tca gtt gca gac att gag cga ata aat cat gaa 1220Asp Asn Gln Lys Tyr
Ser Val Ala Asp Ile Glu Arg Ile Asn His Glu 360 365 370 aga aat gaa
ttg cag cag act att aat aaa tta acc aag gac ctg gaa 1268Arg Asn Glu
Leu Gln Gln Thr Ile Asn Lys Leu Thr Lys Asp Leu Glu 375 380 385 gct
gaa caa cag aag ttg tgg aat gag gag tta aaa tat gcc aga ggc 1316Ala
Glu Gln Gln Lys Leu Trp Asn Glu Glu Leu Lys Tyr Ala Arg Gly 390 395
400 aaa gaa gcg att gaa aca caa tta gca gag tat cac aaa ttg gct aga
1364Lys Glu Ala Ile Glu Thr Gln Leu Ala Glu Tyr His Lys Leu Ala Arg
405 410 415 420 aaa tta aaa ctt att cct aaa ggt gct gag aat tcc aaa
ggt tat gac 1412Lys Leu Lys Leu Ile Pro Lys Gly Ala Glu Asn Ser Lys
Gly Tyr Asp 425 430 435 ttt gaa att aag ttt aat ccc gag gct ggt gcc
aac tgc ctt gtc aaa 1460Phe Glu Ile Lys Phe Asn Pro Glu Ala Gly Ala
Asn Cys Leu Val Lys 440 445 450 tac agg gct caa gtt tat gta cct ctt
aag gaa ctc ctg aat gaa act 1508Tyr Arg Ala Gln Val Tyr Val Pro Leu
Lys Glu Leu Leu Asn Glu Thr 455 460 465 gaa gaa gaa att aat aaa gcc
cta aat aaa aaa atg ggt ttg gag gat 1556Glu Glu Glu Ile Asn Lys Ala
Leu Asn Lys Lys Met Gly Leu Glu Asp 470 475 480 act tta gaa caa ttg
aat gca atg ata aca gaa agc aag aga agt gtg 1604Thr Leu Glu Gln Leu
Asn Ala Met Ile Thr Glu Ser Lys Arg Ser Val 485 490 495 500 aga act
ctg aaa gaa gaa gtt caa aag ctg gat gat ctt tac caa caa 1652Arg Thr
Leu Lys Glu Glu Val Gln Lys Leu Asp Asp Leu Tyr Gln Gln 505 510 515
aaa att aag gaa gca gag gaa gag gat gaa aaa tgt gcc agt gag ctt
1700Lys Ile Lys Glu Ala Glu Glu Glu Asp Glu Lys Cys Ala Ser Glu Leu
520 525 530 gag tcc ttg gag aaa cac aag cac ctg cta gaa agt act gtt
aac cag 1748Glu Ser Leu Glu Lys His Lys His Leu Leu Glu Ser Thr Val
Asn Gln 535 540 545 ggg ctc agt gaa gct atg aat gaa tta gat gct gtt
cag cgg gaa tac 1796Gly Leu Ser Glu Ala Met Asn Glu Leu Asp Ala Val
Gln Arg Glu Tyr 550 555 560 caa cta gtt gtg caa acc acg act gaa gaa
aga cga aaa gtg gga aat 1844Gln Leu Val Val Gln Thr Thr Thr Glu Glu
Arg Arg Lys Val Gly Asn 565 570 575 580 aac ttg caa cgt ctg tta gag
atg gtt gct aca cat gtt ggg tct gta 1892Asn Leu Gln Arg Leu Leu Glu
Met Val Ala Thr His Val Gly Ser Val 585 590 595 gag aaa cat ctt gag
gag cag att gct aaa gtt gat aga gaa tat gaa 1940Glu Lys His Leu Glu
Glu Gln Ile Ala Lys Val Asp Arg Glu Tyr Glu 600 605 610 gaa tgc atg
tca gaa gat ctc tcg gaa aat att aaa gag att aga gat 1988Glu Cys Met
Ser Glu Asp Leu Ser Glu Asn Ile Lys Glu Ile Arg Asp 615 620 625 aag
tat gag aag aaa gct act cta att aag tct tct gaa gaa tga 2033Lys Tyr
Glu Lys Lys Ala Thr Leu Ile Lys Ser Ser Glu Glu 630 635 640
agataaaatg ttgatcatgt atatatatcc atagtgaata aaattgtctc agtaaaaaaa
2093aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa
215014642PRTHomo sapiens 14Met Lys Arg Ser Ser Val Ser Ser Gly Gly
Ala Gly Arg Leu Ser Met 1 5 10 15 Gln Glu Leu Arg Ser Gln Asp Val
Asn Lys Gln Gly Leu Tyr Thr Pro 20 25 30 Gln Thr Lys Glu Lys Pro
Thr Phe Gly Lys Leu Ser Ile Asn Lys Pro 35 40 45 Thr Ser Glu Arg
Lys Val Ser Leu Phe Gly Lys Arg Thr Ser Gly His 50 55 60 Gly Ser
Arg Asn Ser Gln Leu Gly Ile Phe Ser Ser Ser Glu Lys Ile 65 70 75 80
Lys Asp Pro Arg Pro Leu Asn Asp Lys Ala Phe Ile Gln Gln Cys Ile 85
90 95 Arg Gln Leu Cys Glu Phe Leu Thr Glu Asn Gly Tyr Ala His Asn
Val 100 105 110 Ser Met Lys Ser Leu Gln Ala Pro Ser Val Lys Asp Phe
Leu Lys Ile 115 120 125 Phe Thr Phe Leu Tyr Gly Phe Leu Cys Pro Ser
Tyr Glu Leu Pro Asp 130 135 140 Thr Lys Phe Glu Glu Glu Val Pro Arg
Ile Phe Lys Asp Leu Gly Tyr 145 150 155 160 Pro Phe Ala Leu Ser Lys
Ser Ser Met Tyr Thr Val Gly Ala Pro His 165 170 175 Thr Trp Pro His
Ile Val Ala Ala Leu Val Trp Leu Ile Asp Cys Ile 180 185 190 Lys Ile
His Thr Ala Met Lys Glu Ser Ser Pro Leu Phe Asp Asp Gly 195 200 205
Gln Pro Trp Gly Glu Glu Thr Glu Asp Gly Ile Met His Asn Lys Leu 210
215 220 Phe Leu Asp Tyr Thr Ile Lys Cys Tyr Glu Ser Phe Met Ser Gly
Ala 225 230 235 240 Asp Ser Phe Asp Glu Met Asn Ala Glu Leu Gln Ser
Lys Leu Lys Asp 245 250 255 Leu Phe Asn Val Asp Ala Phe Lys Leu Glu
Ser Leu Glu Ala Lys Asn 260 265 270 Arg Ala Leu Asn Glu Gln Ile Ala
Arg Leu Glu Gln Glu Arg Glu Lys 275 280 285 Glu Pro Asn Arg Leu Glu
Ser Leu Arg Lys Leu Lys Ala Ser Leu Gln 290 295 300 Gly Asp Val Gln
Lys Tyr Gln Ala Tyr Met Ser Asn Leu Glu Ser His 305 310 315 320 Ser
Ala Ile Leu Asp Gln Lys Leu Asn Gly Leu Asn Glu Glu Ile Ala 325 330
335 Arg Val Glu Leu Glu Cys Glu Thr Ile Lys Gln Glu Asn Thr Arg Leu
340 345 350 Gln Asn Ile Ile Asp Asn Gln Lys Tyr Ser Val Ala Asp Ile
Glu Arg 355 360 365 Ile Asn His Glu Arg Asn Glu Leu Gln Gln Thr Ile
Asn Lys Leu Thr 370 375 380 Lys Asp Leu Glu Ala Glu Gln Gln Lys Leu
Trp Asn Glu Glu Leu Lys 385 390 395 400 Tyr Ala Arg
Gly Lys Glu Ala Ile Glu Thr Gln Leu Ala Glu Tyr His 405 410 415 Lys
Leu Ala Arg Lys Leu Lys Leu Ile Pro Lys Gly Ala Glu Asn Ser 420 425
430 Lys Gly Tyr Asp Phe Glu Ile Lys Phe Asn Pro Glu Ala Gly Ala Asn
435 440 445 Cys Leu Val Lys Tyr Arg Ala Gln Val Tyr Val Pro Leu Lys
Glu Leu 450 455 460 Leu Asn Glu Thr Glu Glu Glu Ile Asn Lys Ala Leu
Asn Lys Lys Met 465 470 475 480 Gly Leu Glu Asp Thr Leu Glu Gln Leu
Asn Ala Met Ile Thr Glu Ser 485 490 495 Lys Arg Ser Val Arg Thr Leu
Lys Glu Glu Val Gln Lys Leu Asp Asp 500 505 510 Leu Tyr Gln Gln Lys
Ile Lys Glu Ala Glu Glu Glu Asp Glu Lys Cys 515 520 525 Ala Ser Glu
Leu Glu Ser Leu Glu Lys His Lys His Leu Leu Glu Ser 530 535 540 Thr
Val Asn Gln Gly Leu Ser Glu Ala Met Asn Glu Leu Asp Ala Val 545 550
555 560 Gln Arg Glu Tyr Gln Leu Val Val Gln Thr Thr Thr Glu Glu Arg
Arg 565 570 575 Lys Val Gly Asn Asn Leu Gln Arg Leu Leu Glu Met Val
Ala Thr His 580 585 590 Val Gly Ser Val Glu Lys His Leu Glu Glu Gln
Ile Ala Lys Val Asp 595 600 605 Arg Glu Tyr Glu Glu Cys Met Ser Glu
Asp Leu Ser Glu Asn Ile Lys 610 615 620 Glu Ile Arg Asp Lys Tyr Glu
Lys Lys Ala Thr Leu Ile Lys Ser Ser 625 630 635 640 Glu Glu
152984DNAHomo sapiensCDS(75)..(2648) 15ggaaattcaa acgtgtttgc
ggaaaggagt ttgggttcca tcttttcatt tccccagcgc 60agctttctgt agaa atg
gaa tcc gag gat tta agt ggc aga gaa ttg aca 110 Met Glu Ser Glu Asp
Leu Ser Gly Arg Glu Leu Thr 1 5 10 att gat tcc ata atg aac aaa gtg
aga gac att aaa aat aag ttt aaa 158Ile Asp Ser Ile Met Asn Lys Val
Arg Asp Ile Lys Asn Lys Phe Lys 15 20 25 aat gaa gac ctt act gat
gaa cta agc ttg aat aaa att tct gct gat 206Asn Glu Asp Leu Thr Asp
Glu Leu Ser Leu Asn Lys Ile Ser Ala Asp 30 35 40 act aca gat aac
tcg gga act gtt aac caa att atg atg atg gca aac 254Thr Thr Asp Asn
Ser Gly Thr Val Asn Gln Ile Met Met Met Ala Asn 45 50 55 60 aac cca
gag gac tgg ttg agt ttg ttg ctc aaa cta gag aaa aac agt 302Asn Pro
Glu Asp Trp Leu Ser Leu Leu Leu Lys Leu Glu Lys Asn Ser 65 70 75
gtt ccg cta agt gat gct ctt tta aat aaa ttg att ggt cgt tac agt
350Val Pro Leu Ser Asp Ala Leu Leu Asn Lys Leu Ile Gly Arg Tyr Ser
80 85 90 caa gca att gaa gcg ctt ccc cca gat aaa tat ggc caa aat
gag agt 398Gln Ala Ile Glu Ala Leu Pro Pro Asp Lys Tyr Gly Gln Asn
Glu Ser 95 100 105 ttt gct aga att caa gtg aga ttt gct gaa tta aaa
gct att caa gag 446Phe Ala Arg Ile Gln Val Arg Phe Ala Glu Leu Lys
Ala Ile Gln Glu 110 115 120 cca gat gat gca cgt gac tac ttt caa atg
gcc aga gca aac tgc aag 494Pro Asp Asp Ala Arg Asp Tyr Phe Gln Met
Ala Arg Ala Asn Cys Lys 125 130 135 140 aaa ttt gct ttt gtt cat ata
tct ttt gca caa ttt gaa ctg tca caa 542Lys Phe Ala Phe Val His Ile
Ser Phe Ala Gln Phe Glu Leu Ser Gln 145 150 155 ggt aat gtc aaa aaa
agt aaa caa ctt ctt caa aaa gct gta gaa cgt 590Gly Asn Val Lys Lys
Ser Lys Gln Leu Leu Gln Lys Ala Val Glu Arg 160 165 170 gga gca gta
cca cta gaa atg ctg gaa att gcc ctg cgg aat tta aac 638Gly Ala Val
Pro Leu Glu Met Leu Glu Ile Ala Leu Arg Asn Leu Asn 175 180 185 ctc
caa aaa aag cag ctg ctt tca gag gag gaa aag aag aat tta tca 686Leu
Gln Lys Lys Gln Leu Leu Ser Glu Glu Glu Lys Lys Asn Leu Ser 190 195
200 gca tct acg gta tta act gcc caa gaa tca ttt tcc ggt tca ctt ggg
734Ala Ser Thr Val Leu Thr Ala Gln Glu Ser Phe Ser Gly Ser Leu Gly
205 210 215 220 cat tta cag aat agg aac aac agt tgt gat tcc aga gga
cag act act 782His Leu Gln Asn Arg Asn Asn Ser Cys Asp Ser Arg Gly
Gln Thr Thr 225 230 235 aaa gcc agg ttt tta tat gga gag aac atg cca
cca caa gat gca gaa 830Lys Ala Arg Phe Leu Tyr Gly Glu Asn Met Pro
Pro Gln Asp Ala Glu 240 245 250 ata ggt tac cgg aat tca ttg aga caa
act aac aaa act aaa cag tca 878Ile Gly Tyr Arg Asn Ser Leu Arg Gln
Thr Asn Lys Thr Lys Gln Ser 255 260 265 tgc cca ttt gga aga gtc cca
gtt aac ctt cta aat agc cca gat tgt 926Cys Pro Phe Gly Arg Val Pro
Val Asn Leu Leu Asn Ser Pro Asp Cys 270 275 280 gat gtg aag aca gat
gat tca gtt gta cct tgt ttt atg aaa aga caa 974Asp Val Lys Thr Asp
Asp Ser Val Val Pro Cys Phe Met Lys Arg Gln 285 290 295 300 acc tct
aga tca gaa tgc cga gat ttg gtt gtg cct gga tct aaa cca 1022Thr Ser
Arg Ser Glu Cys Arg Asp Leu Val Val Pro Gly Ser Lys Pro 305 310 315
agt gga aat gat tcc tgt gaa tta aga aat tta aag tct gtt caa aat
1070Ser Gly Asn Asp Ser Cys Glu Leu Arg Asn Leu Lys Ser Val Gln Asn
320 325 330 agt cat ttc aag gaa cct ctg gtg tca gat gaa aag agt tct
gaa ctt 1118Ser His Phe Lys Glu Pro Leu Val Ser Asp Glu Lys Ser Ser
Glu Leu 335 340 345 att att act gat tca ata acc ctg aag aat aaa acg
gaa tca agt ctt 1166Ile Ile Thr Asp Ser Ile Thr Leu Lys Asn Lys Thr
Glu Ser Ser Leu 350 355 360 cta gct aaa tta gaa gaa act aaa gag tat
caa gaa cca gag gtt cca 1214Leu Ala Lys Leu Glu Glu Thr Lys Glu Tyr
Gln Glu Pro Glu Val Pro 365 370 375 380 gag agt aac cag aaa cag tgg
caa tct aag aga aag tca gag tgt att 1262Glu Ser Asn Gln Lys Gln Trp
Gln Ser Lys Arg Lys Ser Glu Cys Ile 385 390 395 aac cag aat cct gct
gca tct tca aat cac tgg cag att ccg gag tta 1310Asn Gln Asn Pro Ala
Ala Ser Ser Asn His Trp Gln Ile Pro Glu Leu 400 405 410 gcc cga aaa
gtt aat aca gag cag aaa cat acc act ttt gag caa cct 1358Ala Arg Lys
Val Asn Thr Glu Gln Lys His Thr Thr Phe Glu Gln Pro 415 420 425 gtc
ttt tca gtt tca aaa cag tca cca cca ata tca aca tct aaa tgg 1406Val
Phe Ser Val Ser Lys Gln Ser Pro Pro Ile Ser Thr Ser Lys Trp 430 435
440 ttt gac cca aaa tct att tgt aag aca cca agc agc aat acc ttg gat
1454Phe Asp Pro Lys Ser Ile Cys Lys Thr Pro Ser Ser Asn Thr Leu Asp
445 450 455 460 gat tac atg agc tgt ttt aga act cca gtt gta aag aat
gac ttt cca 1502Asp Tyr Met Ser Cys Phe Arg Thr Pro Val Val Lys Asn
Asp Phe Pro 465 470 475 cct gct tgt cag ttg tca aca cct tat ggc caa
cct gcc tgt ttc cag 1550Pro Ala Cys Gln Leu Ser Thr Pro Tyr Gly Gln
Pro Ala Cys Phe Gln 480 485 490 cag caa cag cat caa ata ctt gcc act
cca ctt caa aat tta cag gtt 1598Gln Gln Gln His Gln Ile Leu Ala Thr
Pro Leu Gln Asn Leu Gln Val 495 500 505 tta gca tct tct tca gca aat
gaa tgc att tcg gtt aaa gga aga att 1646Leu Ala Ser Ser Ser Ala Asn
Glu Cys Ile Ser Val Lys Gly Arg Ile 510 515 520 tat tcc att tta aag
cag ata gga agt gga ggt tca agc aag gta ttt 1694Tyr Ser Ile Leu Lys
Gln Ile Gly Ser Gly Gly Ser Ser Lys Val Phe 525 530 535 540 cag gtg
tta aat gaa aag aaa cag ata tat gct ata aaa tat gtg aac 1742Gln Val
Leu Asn Glu Lys Lys Gln Ile Tyr Ala Ile Lys Tyr Val Asn 545 550 555
tta gaa gaa gca gat aac caa act ctt gat agt tac cgg aac gaa ata
1790Leu Glu Glu Ala Asp Asn Gln Thr Leu Asp Ser Tyr Arg Asn Glu Ile
560 565 570 gct tat ttg aat aaa cta caa caa cac agt gat aag atc atc
cga ctt 1838Ala Tyr Leu Asn Lys Leu Gln Gln His Ser Asp Lys Ile Ile
Arg Leu 575 580 585 tat gat tat gaa atc acg gac cag tac atc tac atg
gta atg gag tgt 1886Tyr Asp Tyr Glu Ile Thr Asp Gln Tyr Ile Tyr Met
Val Met Glu Cys 590 595 600 gga aat att gat ctt aat agt tgg ctt aaa
aag aaa aaa tcc att gat 1934Gly Asn Ile Asp Leu Asn Ser Trp Leu Lys
Lys Lys Lys Ser Ile Asp 605 610 615 620 cca tgg gaa cgc aag agt tac
tgg aaa aat atg tta gag gca gtt cac 1982Pro Trp Glu Arg Lys Ser Tyr
Trp Lys Asn Met Leu Glu Ala Val His 625 630 635 aca atc cat caa cat
ggc att gtt cac agt gat ctt aaa cca gct aac 2030Thr Ile His Gln His
Gly Ile Val His Ser Asp Leu Lys Pro Ala Asn 640 645 650 ttt ctg ata
gtt gat gga atg cta aag cta att gat ttt ggg att gca 2078Phe Leu Ile
Val Asp Gly Met Leu Lys Leu Ile Asp Phe Gly Ile Ala 655 660 665 aac
caa atg caa cca gat aca aca agt gtt gtt aaa gat tct cag gtt 2126Asn
Gln Met Gln Pro Asp Thr Thr Ser Val Val Lys Asp Ser Gln Val 670 675
680 ggc aca gtt aat tat atg cca cca gaa gca atc aaa gat atg tct tcc
2174Gly Thr Val Asn Tyr Met Pro Pro Glu Ala Ile Lys Asp Met Ser Ser
685 690 695 700 tcc aga gag aat ggg aaa tct aag tca aag ata agc ccc
aaa agt gat 2222Ser Arg Glu Asn Gly Lys Ser Lys Ser Lys Ile Ser Pro
Lys Ser Asp 705 710 715 gtt tgg tcc tta gga tgt att ttg tac tat atg
act tac ggg aaa aca 2270Val Trp Ser Leu Gly Cys Ile Leu Tyr Tyr Met
Thr Tyr Gly Lys Thr 720 725 730 cca ttt cag cag ata att aat cag att
tct aaa tta cat gcc ata att 2318Pro Phe Gln Gln Ile Ile Asn Gln Ile
Ser Lys Leu His Ala Ile Ile 735 740 745 gat cct aat cat gaa att gaa
ttt ccc gat att cca gag aaa gat ctt 2366Asp Pro Asn His Glu Ile Glu
Phe Pro Asp Ile Pro Glu Lys Asp Leu 750 755 760 caa gat gtg tta aag
tgt tgt tta aaa agg gac cca aaa cag agg ata 2414Gln Asp Val Leu Lys
Cys Cys Leu Lys Arg Asp Pro Lys Gln Arg Ile 765 770 775 780 tcc att
cct gag ctc ctg gct cat ccc tat gtt caa att caa act cat 2462Ser Ile
Pro Glu Leu Leu Ala His Pro Tyr Val Gln Ile Gln Thr His 785 790 795
cca gtt aac caa atg gcc aag gga acc act gaa gaa atg aaa tat gtt
2510Pro Val Asn Gln Met Ala Lys Gly Thr Thr Glu Glu Met Lys Tyr Val
800 805 810 ctg ggc caa ctt gtt ggt ctg aat tct cct aac tcc att ttg
aaa gct 2558Leu Gly Gln Leu Val Gly Leu Asn Ser Pro Asn Ser Ile Leu
Lys Ala 815 820 825 gct aaa act tta tat gaa cac tat agt ggt ggt gaa
agt cat aat tct 2606Ala Lys Thr Leu Tyr Glu His Tyr Ser Gly Gly Glu
Ser His Asn Ser 830 835 840 tca tcc tcc aag act ttt gaa aaa aaa agg
gga aaa aaa tga 2648Ser Ser Ser Lys Thr Phe Glu Lys Lys Arg Gly Lys
Lys 845 850 855 tttgcagtta ttcgtaatgt caaataccac ctataaaata
tattggactg ttatactctt 2708gaatccctgt ggaaatctac atttgaagac
aacatcactc tgaagtgtta tcagcaaaaa 2768aaattcagta gattatcttt
aaaagaaaac tgtaaaaata gcaaccactt atggtactgt 2828atatattgta
gacttgtttt ctctgtttta tgctcttgtg taatctactt gacatcattt
2888tactcttgga atagtgggtg gatagcaagt atattctaaa aaactttgta
aataaagttt 2948tgtggctaaa atgacactaa aaaaaaaaaa aaaaaa
298416857PRTHomo sapiens 16Met Glu Ser Glu Asp Leu Ser Gly Arg Glu
Leu Thr Ile Asp Ser Ile 1 5 10 15 Met Asn Lys Val Arg Asp Ile Lys
Asn Lys Phe Lys Asn Glu Asp Leu 20 25 30 Thr Asp Glu Leu Ser Leu
Asn Lys Ile Ser Ala Asp Thr Thr Asp Asn 35 40 45 Ser Gly Thr Val
Asn Gln Ile Met Met Met Ala Asn Asn Pro Glu Asp 50 55 60 Trp Leu
Ser Leu Leu Leu Lys Leu Glu Lys Asn Ser Val Pro Leu Ser 65 70 75 80
Asp Ala Leu Leu Asn Lys Leu Ile Gly Arg Tyr Ser Gln Ala Ile Glu 85
90 95 Ala Leu Pro Pro Asp Lys Tyr Gly Gln Asn Glu Ser Phe Ala Arg
Ile 100 105 110 Gln Val Arg Phe Ala Glu Leu Lys Ala Ile Gln Glu Pro
Asp Asp Ala 115 120 125 Arg Asp Tyr Phe Gln Met Ala Arg Ala Asn Cys
Lys Lys Phe Ala Phe 130 135 140 Val His Ile Ser Phe Ala Gln Phe Glu
Leu Ser Gln Gly Asn Val Lys 145 150 155 160 Lys Ser Lys Gln Leu Leu
Gln Lys Ala Val Glu Arg Gly Ala Val Pro 165 170 175 Leu Glu Met Leu
Glu Ile Ala Leu Arg Asn Leu Asn Leu Gln Lys Lys 180 185 190 Gln Leu
Leu Ser Glu Glu Glu Lys Lys Asn Leu Ser Ala Ser Thr Val 195 200 205
Leu Thr Ala Gln Glu Ser Phe Ser Gly Ser Leu Gly His Leu Gln Asn 210
215 220 Arg Asn Asn Ser Cys Asp Ser Arg Gly Gln Thr Thr Lys Ala Arg
Phe 225 230 235 240 Leu Tyr Gly Glu Asn Met Pro Pro Gln Asp Ala Glu
Ile Gly Tyr Arg 245 250 255 Asn Ser Leu Arg Gln Thr Asn Lys Thr Lys
Gln Ser Cys Pro Phe Gly 260 265 270 Arg Val Pro Val Asn Leu Leu Asn
Ser Pro Asp Cys Asp Val Lys Thr 275 280 285 Asp Asp Ser Val Val Pro
Cys Phe Met Lys Arg Gln Thr Ser Arg Ser 290 295 300 Glu Cys Arg Asp
Leu Val Val Pro Gly Ser Lys Pro Ser Gly Asn Asp 305 310 315 320 Ser
Cys Glu Leu Arg Asn Leu Lys Ser Val Gln Asn Ser His Phe Lys 325 330
335 Glu Pro Leu Val Ser Asp Glu Lys Ser Ser Glu Leu Ile Ile Thr Asp
340 345 350 Ser Ile Thr Leu Lys Asn Lys Thr Glu Ser Ser Leu Leu Ala
Lys Leu 355 360 365 Glu Glu Thr Lys Glu Tyr Gln Glu Pro Glu Val Pro
Glu Ser Asn Gln 370 375 380 Lys Gln Trp Gln Ser Lys Arg Lys Ser Glu
Cys Ile Asn Gln Asn Pro 385 390 395 400 Ala Ala Ser Ser Asn His Trp
Gln Ile Pro Glu Leu Ala Arg Lys Val 405 410 415 Asn Thr Glu Gln Lys
His Thr Thr Phe Glu Gln Pro Val Phe Ser Val 420 425 430 Ser Lys Gln
Ser Pro Pro Ile Ser Thr Ser Lys Trp Phe Asp Pro Lys 435 440 445 Ser
Ile Cys Lys Thr Pro Ser Ser Asn Thr Leu Asp Asp Tyr Met Ser 450 455
460 Cys Phe Arg Thr Pro Val Val Lys Asn Asp Phe Pro Pro Ala Cys Gln
465 470 475 480 Leu Ser Thr Pro Tyr Gly Gln Pro Ala Cys Phe Gln Gln
Gln Gln His 485 490 495 Gln Ile Leu Ala Thr Pro Leu Gln Asn Leu Gln
Val Leu Ala Ser Ser 500 505 510
Ser Ala Asn Glu Cys Ile Ser Val Lys Gly Arg Ile Tyr Ser Ile Leu 515
520 525 Lys Gln Ile Gly Ser Gly Gly Ser Ser Lys Val Phe Gln Val Leu
Asn 530 535 540 Glu Lys Lys Gln Ile Tyr Ala Ile Lys Tyr Val Asn Leu
Glu Glu Ala 545 550 555 560 Asp Asn Gln Thr Leu Asp Ser Tyr Arg Asn
Glu Ile Ala Tyr Leu Asn 565 570 575 Lys Leu Gln Gln His Ser Asp Lys
Ile Ile Arg Leu Tyr Asp Tyr Glu 580 585 590 Ile Thr Asp Gln Tyr Ile
Tyr Met Val Met Glu Cys Gly Asn Ile Asp 595 600 605 Leu Asn Ser Trp
Leu Lys Lys Lys Lys Ser Ile Asp Pro Trp Glu Arg 610 615 620 Lys Ser
Tyr Trp Lys Asn Met Leu Glu Ala Val His Thr Ile His Gln 625 630 635
640 His Gly Ile Val His Ser Asp Leu Lys Pro Ala Asn Phe Leu Ile Val
645 650 655 Asp Gly Met Leu Lys Leu Ile Asp Phe Gly Ile Ala Asn Gln
Met Gln 660 665 670 Pro Asp Thr Thr Ser Val Val Lys Asp Ser Gln Val
Gly Thr Val Asn 675 680 685 Tyr Met Pro Pro Glu Ala Ile Lys Asp Met
Ser Ser Ser Arg Glu Asn 690 695 700 Gly Lys Ser Lys Ser Lys Ile Ser
Pro Lys Ser Asp Val Trp Ser Leu 705 710 715 720 Gly Cys Ile Leu Tyr
Tyr Met Thr Tyr Gly Lys Thr Pro Phe Gln Gln 725 730 735 Ile Ile Asn
Gln Ile Ser Lys Leu His Ala Ile Ile Asp Pro Asn His 740 745 750 Glu
Ile Glu Phe Pro Asp Ile Pro Glu Lys Asp Leu Gln Asp Val Leu 755 760
765 Lys Cys Cys Leu Lys Arg Asp Pro Lys Gln Arg Ile Ser Ile Pro Glu
770 775 780 Leu Leu Ala His Pro Tyr Val Gln Ile Gln Thr His Pro Val
Asn Gln 785 790 795 800 Met Ala Lys Gly Thr Thr Glu Glu Met Lys Tyr
Val Leu Gly Gln Leu 805 810 815 Val Gly Leu Asn Ser Pro Asn Ser Ile
Leu Lys Ala Ala Lys Thr Leu 820 825 830 Tyr Glu His Tyr Ser Gly Gly
Glu Ser His Asn Ser Ser Ser Ser Lys 835 840 845 Thr Phe Glu Lys Lys
Arg Gly Lys Lys 850 855 171735DNAHomo sapiensCDS(242)..(913)
17gttatcagag gtgagcccgt gctcttcagc ggagaagatc ccctacctgg ccgccggcca
60ctttctgtgg gccgtggggt cctcaaggag acggcccttg ggctcagggg ctgcgtttcc
120acacgcgcct ttcccagggc tcccgcgccc gttcctgcct ggccgccggc
cgctccaaca 180gcagcacaag gcgggactca gaaccggcgt tcagggccgc
cagcggccgc gaggccctga 240g atg agg ctc caa aga ccc cga cag gcc ccg
gcg ggt ggg agg cgc gcg 289 Met Arg Leu Gln Arg Pro Arg Gln Ala Pro
Ala Gly Gly Arg Arg Ala 1 5 10 15 ccc cgg ggc ggg cgg ggc tcc ccc
tac cgg cca gac ccg ggg aga ggc 337Pro Arg Gly Gly Arg Gly Ser Pro
Tyr Arg Pro Asp Pro Gly Arg Gly 20 25 30 gcg cgg agg ctg cga agg
ttc cag aag ggc ggg gag ggg gcg ccg cgc 385Ala Arg Arg Leu Arg Arg
Phe Gln Lys Gly Gly Glu Gly Ala Pro Arg 35 40 45 gct gac cct ccc
tgg gca ccg ctg ggg acg atg gcg ctg ctc gcc ttg 433Ala Asp Pro Pro
Trp Ala Pro Leu Gly Thr Met Ala Leu Leu Ala Leu 50 55 60 ctg ctg
gtc gtg gcc cta ccg cgg gtg tgg aca gac gcc aac ctg act 481Leu Leu
Val Val Ala Leu Pro Arg Val Trp Thr Asp Ala Asn Leu Thr 65 70 75 80
gcg aga caa cga gat cca gag gac tcc cag cga acg gac gag ggt gac
529Ala Arg Gln Arg Asp Pro Glu Asp Ser Gln Arg Thr Asp Glu Gly Asp
85 90 95 aat aga gtg tgg tgt cat gtt tgt gag aga gaa aac act ttc
gag tgc 577Asn Arg Val Trp Cys His Val Cys Glu Arg Glu Asn Thr Phe
Glu Cys 100 105 110 cag aac cca agg agg tgc aaa tgg aca gag cca tac
tgc gtt ata gcg 625Gln Asn Pro Arg Arg Cys Lys Trp Thr Glu Pro Tyr
Cys Val Ile Ala 115 120 125 gcc gtg aaa ata ttt cca cgt ttt ttc atg
gtt gcg aag cag tgc tcc 673Ala Val Lys Ile Phe Pro Arg Phe Phe Met
Val Ala Lys Gln Cys Ser 130 135 140 gct ggt tgt gca gcg atg gag aga
ccc aag cca gag gag aag cgg ttt 721Ala Gly Cys Ala Ala Met Glu Arg
Pro Lys Pro Glu Glu Lys Arg Phe 145 150 155 160 ctc ctg gaa gag ccc
atg ccc ttc ttt tac ctc aag tgt tgt aaa att 769Leu Leu Glu Glu Pro
Met Pro Phe Phe Tyr Leu Lys Cys Cys Lys Ile 165 170 175 cgc tac tgc
aat tta gag ggg cca cct atc aac tca tca gtg ttc aaa 817Arg Tyr Cys
Asn Leu Glu Gly Pro Pro Ile Asn Ser Ser Val Phe Lys 180 185 190 gaa
tat gct ggg agc atg ggt gag agc tgt ggt ggg ctg tgg ctg gcc 865Glu
Tyr Ala Gly Ser Met Gly Glu Ser Cys Gly Gly Leu Trp Leu Ala 195 200
205 atc ctc ctg ctg ctg gcc tcc att gca gcc ggc ctc agc ctg tct tga
913Ile Leu Leu Leu Leu Ala Ser Ile Ala Ala Gly Leu Ser Leu Ser 210
215 220 gccacgggac tgccacagac tgagccttcc ggagcatgga ctcgctccag
accgttgtca 973cctgttgcat taaacttgtt ttctgttgat tacctcttgg
tttgacttcc cagggtcttg 1033ggatgggaga gtggggatca ggtgcagttg
gctcttaacc ctcaagggtt ctttaactca 1093cattcagagg aagtccagat
ctcctgagta gtgattttgg tgacaagttt ttctctttga 1153aatcaaacct
tgtaactcat ttattgctga tggccactct tttccttgac tcccctctgc
1213ctctgagggc ttcagtattg atggggaggg aggcctaagt accactcatg
gagagtatgt 1273gctgagatgc ttccgacctt tcaggtgacg caggaacact
gggggagtct gaatgattgg 1333ggtgaagaca tccctggagt gaaggactcc
tcagcatggg gggcagtggg gcacacgtta 1393gggctgcccc cattccagtg
gtggaggcgc tgtggatggc tgcttttcct caacctttcc 1453taccagattc
caggaggcag aagataacta attgtgttga agaaacttag acttcaccca
1513ccagctggca caggtgcaca gattcataaa ttcccacacg tgtgtgttca
acatctgaaa 1573cttaggccaa gtagagagca tcagggtaaa tggcgttcat
ttctctgtta agatgcagcc 1633atccatgggg agctgagaaa tcagactcaa
agttccacca aaaacaaata caaggggact 1693tcaaaagttc acgaaaaaat
tgaattaaaa gataaaaatt aa 173518223PRTHomo sapiens 18Met Arg Leu Gln
Arg Pro Arg Gln Ala Pro Ala Gly Gly Arg Arg Ala 1 5 10 15 Pro Arg
Gly Gly Arg Gly Ser Pro Tyr Arg Pro Asp Pro Gly Arg Gly 20 25 30
Ala Arg Arg Leu Arg Arg Phe Gln Lys Gly Gly Glu Gly Ala Pro Arg 35
40 45 Ala Asp Pro Pro Trp Ala Pro Leu Gly Thr Met Ala Leu Leu Ala
Leu 50 55 60 Leu Leu Val Val Ala Leu Pro Arg Val Trp Thr Asp Ala
Asn Leu Thr 65 70 75 80 Ala Arg Gln Arg Asp Pro Glu Asp Ser Gln Arg
Thr Asp Glu Gly Asp 85 90 95 Asn Arg Val Trp Cys His Val Cys Glu
Arg Glu Asn Thr Phe Glu Cys 100 105 110 Gln Asn Pro Arg Arg Cys Lys
Trp Thr Glu Pro Tyr Cys Val Ile Ala 115 120 125 Ala Val Lys Ile Phe
Pro Arg Phe Phe Met Val Ala Lys Gln Cys Ser 130 135 140 Ala Gly Cys
Ala Ala Met Glu Arg Pro Lys Pro Glu Glu Lys Arg Phe 145 150 155 160
Leu Leu Glu Glu Pro Met Pro Phe Phe Tyr Leu Lys Cys Cys Lys Ile 165
170 175 Arg Tyr Cys Asn Leu Glu Gly Pro Pro Ile Asn Ser Ser Val Phe
Lys 180 185 190 Glu Tyr Ala Gly Ser Met Gly Glu Ser Cys Gly Gly Leu
Trp Leu Ala 195 200 205 Ile Leu Leu Leu Leu Ala Ser Ile Ala Ala Gly
Leu Ser Leu Ser 210 215 220 199PRTartificialAn artificially
synthesized peptide sequence 19Ile Tyr Glu Val Met Val Leu Ala Met
1 5 209PRTartificialAn artificially synthesized peptide sequence
20Leu Phe Leu Leu Leu Val Leu Leu Leu 1 5 219PRTartificialAn
artificially synthesized peptide sequence 21Val Phe Arg Glu Ala Glu
Val Thr Leu 1 5 229PRTartificialAn artificially synthesized peptide
sequence 22Leu Tyr Val Glu Val Thr Asn Glu Ala 1 5
239PRTArtificialAn artificially synthesized peptide sequence 23Lys
Tyr Glu Ala His Val Pro Glu Asn 1 5 249PRTArtificialAn artificially
synthesized peptide sequence 24Lys Tyr Glu Leu Phe Gly His Ala Val
1 5 259PRTArtificialAn artificially synthesized peptide sequence
25Arg Ser Leu Lys Glu Arg Asn Pro Leu 1 5 269PRTArtificialAn
artificially synthesized peptide sequence 26Arg Gly Pro Leu Ala Ser
Leu Leu Leu 1 5 279PRTArtificialAn artificially synthesized peptide
sequence 27Lys Gly Gly Phe Ile Leu Pro Val Leu 1 5
289PRTArtificialAn artificially synthesized peptide sequence 28Thr
Tyr Asn Gly Val Val Ala Tyr Ser 1 5 299PRTArtificialAn artificially
synthesized peptide sequence 29Leu Phe Ser Thr Asp Asn Asp Asp Phe
1 5 3010PRTArtificialAn artificially synthesized peptide sequence
30Asp Tyr Leu Asn Glu Trp Gly Ser Arg Phe 1 5 10
3110PRTArtificialAn artificially synthesized peptide sequence 31Thr
Tyr Asn Gly Val Val Ala Tyr Ser Ile 1 5 10 3210PRTArtificialAn
artificially synthesized peptide sequence 32Leu Phe Leu Leu Leu Val
Leu Leu Leu Leu 1 5 10 3310PRTArtificialAn artificially synthesized
peptide sequence 33Asp Phe Glu Ala Lys Asn Gln His Thr Leu 1 5 10
3410PRTArtificialAn artificially synthesized peptide sequence 34Lys
Tyr Glu Ala His Val Pro Glu Asn Ala 1 5 10 3510PRTArtificialAn
artificially synthesized peptide sequence 35Lys Tyr Glu Leu Phe Gly
His Ala Val Ser 1 5 10 3610PRTArtificialAn artificially synthesized
peptide sequence 36Arg Asn Asn Ile Tyr Glu Val Met Val Leu 1 5 10
3710PRTArtificialAn artificially synthesized peptide sequence 37Arg
Gly Pro Leu Ala Ser Leu Leu Leu Leu 1 5 10 3810PRTArtificialAn
artificially synthesized peptide sequence 38Arg Ile Leu Arg Asp Pro
Ala Gly Trp Leu 1 5 10 3910PRTArtificialAn artificially synthesized
peptide sequence 39Cys Asn Gln Ser Pro Val Arg Gln Val Leu 1 5 10
409PRTArtificialAn artificially synthesized peptide sequence 40Val
Tyr Ile Glu Ile Lys Phe Thr Leu 1 5 419PRTArtificialAn artificially
synthesized peptide sequence 41Arg Tyr Ser Val Ala Leu Ala Trp Leu
1 5 429PRTArtificialAn artificially synthesized peptide sequence
42Val Tyr Pro Ala Asn Glu Val Thr Leu 1 5 439PRTArtificialAn
artificially synthesized peptide sequence 43His Tyr Thr Pro Gln Gln
Asn Gly Leu 1 5 449PRTArtificialAn artificially synthesized peptide
sequence 44Phe Tyr Phe Ala Leu Phe Ser Cys Leu 1 5
459PRTArtificialAn artificially synthesized peptide sequence 45Gly
Tyr Gly Asp Phe Ser Glu Pro Leu 1 5 469PRTArtificialAn artificially
synthesized peptide sequence 46Lys Phe Gly Gln Ile Val Asn Met Leu
1 5 479PRTArtificialAn artificially synthesized peptide sequence
47Ala Tyr Thr Thr Arg Gly Gly Lys Ile 1 5 489PRTArtificialAn
artificially synthesized peptide sequence 48Lys Tyr Asn Pro Asn Pro
Asp Gln Ser 1 5 499PRTArtificialAn artificially synthesized peptide
sequence 49Arg Asn Ile Leu Val Asn Ser Asn Leu 1 5
509PRTArtificialAn artificially synthesized peptide sequence 50Lys
Tyr Leu Ser Asp Met Ser Tyr Val 1 5 519PRTArtificialAn artificially
synthesized peptide sequence 51Lys Leu Ile Arg Asn Pro Asn Ser Leu
1 5 529PRTArtificialAn artificially synthesized peptide sequence
52Arg Tyr Lys Asp Asn Phe Thr Ala Ala 1 5 539PRTArtificialAn
artificially synthesized peptide sequence 53Lys Ala Ile Glu Glu Gly
Tyr Arg Leu 1 5 549PRTArtificialAn artificially synthesized peptide
sequence 54Lys Tyr Ser Lys Ala Lys Gln Glu Ala 1 5
559PRTArtificialAn artificially synthesized peptide sequence 55Ala
Phe Gln Asp Val Gly Ala Cys Ile 1 5 569PRTArtificialAn artificially
synthesized peptide sequence 56Trp Leu Val Pro Ile Gly Asn Cys Leu
1 5 579PRTArtificialAn artificially synthesized peptide sequence
57Arg Pro Pro Ser Ala Pro Leu Asn Leu 1 5 589PRTArtificialAn
artificially synthesized peptide sequence 58Lys Cys Pro Leu Thr Val
Arg Asn Leu 1 5 599PRTArtificialAn artificially synthesized peptide
sequence 59Ser Tyr Asn Val Val Cys Lys Lys Cys 1 5
6010PRTArtificialAn artificially synthesized peptide sequence 60Val
Tyr Pro Ala Asn Glu Val Thr Leu Leu 1 5 10 6110PRTArtificialAn
artificially synthesized peptide sequence 61Met Tyr Cys Gly Ala Asp
Gly Glu Trp Leu 1 5 10 6210PRTArtificialAn artificially synthesized
peptide sequence 62Gly Tyr Thr Asp Lys Gln Arg Arg Asp Phe 1 5 10
6310PRTArtificialAn artificially synthesized peptide sequence 63Phe
Tyr Phe Ala Leu Phe Ser Cys Leu Phe 1 5 10 6410PRTArtificialAn
artificially synthesized peptide sequence 64Lys Phe Thr Leu Arg Asp
Cys Asn Ser Leu 1 5 10 6510PRTArtificialAn artificially synthesized
peptide sequence 65Ser Tyr Gly Glu Arg Pro Tyr Trp Asp Met 1 5 10
6610PRTArtificialAn artificially synthesized peptide sequence 66Ile
Phe Tyr Phe Ala Leu Phe Ser Cys Leu 1 5 10
6710PRTArtificialSYGIVMWEVM 67Ser Tyr Gly Ile Val Met Trp Glu Val
Met 1 5 10 6810PRTArtificialAn artificially synthesized peptide
sequence 68Glu Phe Gly Glu Val Cys Ser Gly Arg Leu 1 5 10
6910PRTArtificialAn artificially synthesized peptide sequence 69Lys
Tyr Asn Pro Asn Pro Asp Gln Ser Val 1 5 10 7010PRTArtificialAn
artificially synthesized peptide sequence 70Asn Phe Thr Ala Ala Gly
Tyr Thr Thr Leu 1 5 10 7110PRTArtificialAn artificially synthesized
peptide sequence 71Gln Phe Asp His Pro Asn Ile Ile His Leu 1 5 10
7210PRTArtificialAn artificially synthesized peptide sequence 72Ala
Phe Leu Arg Lys Asn Asp Gly Arg Phe 1 5 10 7310PRTArtificialAn
artificially synthesized peptide sequence 73Lys Gln Glu Ala Asp Glu
Glu Lys His Leu 1 5 10 7410PRTArtificialAn artificially synthesized
peptide sequence 74Arg Gly Ile Gly Ser Gly Met Lys Tyr Leu 1 5 10
7510PRTArtificialAn artificially synthesized peptide sequence 75Arg
Val Tyr Ile Glu Ile Lys Phe Thr Leu 1 5 10 7610PRTArtificialAn
artificially synthesized peptide sequence 76Ser Tyr Val Phe His Val
Arg Ala Arg Thr 1 5 10 7710PRTArtificialAn artificially synthesized
peptide sequence 77Glu Trp Leu Val Pro Ile Gly Asn Cys Leu 1 5 10
7810PRTArtificialAn artificially synthesized peptide sequence 78Arg
Val Tyr Pro Ala Asn Glu Val Thr Leu 1 5 10 7910PRTArtificialAn
artificially synthesized peptide sequence 79Glu Tyr Met Glu Asn Gly
Ser Leu Asp Ala 1 5 10 809PRTArtificialAn artificially synthesized
peptide sequence 80Thr Tyr Pro Pro Phe Val Asn Phe Phe 1 5
819PRTArtificialAn artificially synthesized peptide sequence 81Leu
Tyr Cys Thr Ser Met Met Asn Leu 1 5 829PRTArtificialAn artificially
synthesized peptide seque 82Leu Tyr Val Val Lys Gln Glu Trp Phe 1 5
839PRTArtificialAn artificially synthesized peptide sequence
83Asn
Tyr Val Asn Ile Leu Ala Thr Ile 1 5 849PRTArtificialAn artificially
synthesized peptide sequence 84Ile Tyr Thr Ala Asp Pro Glu Ser Phe
1 5 859PRTArtificialAn artificially synthesized peptide sequence
85Leu Tyr Lys Ala Asp Cys Arg Val Ile 1 5 869PRTArtificialAn
artificially synthesized peptide sequence 86Ser Phe Gln Met Thr Ser
Asp Glu Leu 1 5 879PRTArtificialAn artificially synthesized peptide
sequence 87Ile Phe Leu Lys Tyr Ser Lys Asp Leu 1 5
889PRTArtificialAn artificially synthesized peptide sequence 88Phe
Phe Glu Arg Arg Ser His Thr Leu 1 5 899PRTArtificialAn artificially
synthesized peptide sequence 89Asp Phe Asn Ser Lys Val Thr His Leu
1 5 909PRTArtificialAn artificially synthesized peptide sequence
90Lys Gln Glu Glu Leu Ile Lys Ala Leu 1 5 919PRTArtificialAn
artificially synthesized peptide sequence 91Arg Gly Glu Gln Val Thr
Leu Phe Leu 1 5 929PRTArtificialAn artificially synthesized peptide
sequence 92Arg Leu Pro Ser Val Ala Leu Leu Leu 1 5
939PRTArtificialAn artificially synthesized peptide sequence 93Lys
Pro Glu Cys Gly Arg Gln Ser Leu 1 5 949PRTArtificialAn artificially
synthesized peptide sequence 94Ile Phe Gly Ser Ile Pro Asp Ile Phe
1 5 959PRTArtificialAn artificially synthesized peptide sequence
95Arg Val Ile Gly Pro Pro Val Val Leu 1 5 969PRTArtificialAn
artificially synthesized peptide sequence 96Lys Tyr Ser Lys Asp Leu
Val Lys Thr 1 5 979PRTArtificialAn artificially synthesized peptide
sequence 97Asp Phe Tyr Ala Ala Val Asp Asp Phe 1 5
9810PRTArtificialAn artificially synthesized peptide sequence 98Leu
Tyr Glu Lys Ala Asn Thr Pro Glu Leu 1 5 10 9910PRTArtificialAn
artificially synthesized peptide sequence 99Asn Tyr Val Asn Ile Leu
Ala Thr Ile Ile 1 5 10 10010PRTArtificialAn artificially
synthesized peptide sequence 100Ser Tyr Val Glu Glu Glu Met Pro Gln
Ile 1 5 10 10110PRTArtificialAn artificially synthesized peptide
sequence 101Asp Phe Gln Asp Ser Val Phe Asn Asp Leu 1 5 10
10210PRTArtificialAn artificially synthesized peptide sequence
102Ser Phe Phe Glu Arg Arg Ser His Thr Leu 1 5 10
10310PRTArtificialAn artificially synthesized peptide sequence
103Ser Phe Ser Lys Thr Pro Lys Arg Ala Leu 1 5 10
10410PRTArtificialAn artificially synthesized peptide sequence
104Lys Tyr Leu Pro Leu Gly Asp Glu Arg Cys 1 5 10
10510PRTArtificialAn artificially synthesized peptide sequence
105Glu Phe Glu Gly Leu Asp Ser Pro Glu Phe 1 5 10
10610PRTArtificialAn artificially synthesized peptide sequence
106Lys Val Pro Pro Phe Gln Asp Cys Ile Leu 1 5 10
10710PRTArtificialAn artificially synthesized peptide sequence
107Arg Pro Pro Thr Glu Gln Ala Asn Val Leu 1 5 10
10810PRTArtificialAn artificially synthesized peptide sequence
108Lys Tyr Ser Lys Asp Leu Val Lys Thr Tyr 1 5 10
10910PRTArtificialAn artificially synthesized peptide sequence
109Val Val Glu Glu Asn Ile Val Lys Asp Leu 1 5 10
1109PRTArtificialAn artificially synthesized peptide sequence
110Ile Phe Val Arg Val Met Glu Ser Leu 1 5 1119PRTArtificialAn
artificially synthesized peptide sequence 111Arg Val Met Glu Ser
Leu Glu Gly Leu 1 5 11210PRTArtificialAn artificially synthesized
peptide sequence 112Leu Tyr Leu Leu Gly Val Val Leu Thr Leu 1 5 10
11310PRTArtificialAn artificially synthesized peptide sequence
113Arg Val Met Glu Ser Leu Glu Gly Leu Leu 1 5 10
1149PRTArtificialAn artificially synthesized peptide sequence
114Tyr Leu Leu Gly Val Val Leu Thr Leu 1 5 1159PRTArtificialAn
artificially synthesized peptide sequence 115Val Leu Thr Leu Leu
Ser Ile Phe Val 1 5 1169PRTArtificialAn artificially synthesized
peptide sequence 116Thr Leu Leu Ser Ile Phe Val Arg Val 1 5
1179PRTArtificialAn artificially synthesized peptide sequence
117Val Leu Asn Leu Tyr Leu Leu Gly Val 1 5 1189PRTArtificialAn
artificially synthesized peptide sequence 118Leu Leu Gly Val Val
Leu Thr Leu Leu 1 5 1199PRTArtificialAn artificially synthesized
peptide sequence 119Arg Val Met Glu Ser Leu Glu Gly Leu 1 5
1209PRTArtificialAn artificially synthesized peptide sequence
120Asn Leu Tyr Leu Leu Gly Val Val Leu 1 5 12110PRTArtificialAn
artificially synthesized peptide sequence 121Tyr Leu Leu Gly Val
Val Leu Thr Leu Leu 1 5 10 12210PRTArtificialAn artificially
synthesized peptide sequence 122Val Val Leu Thr Leu Leu Ser Ile Phe
Val 1 5 10 12310PRTArtificialAn artificially synthesized peptide
sequence 123Gly Leu Leu Glu Ser Pro Ser Pro Gly Thr 1 5 10
12410PRTArtificialAn artificially synthesized peptide sequence
124Asn Leu Tyr Leu Leu Gly Val Val Leu Thr 1 5 10
12510PRTArtificialAn artificially synthesized peptide sequence
125Val Leu Asn Leu Tyr Leu Leu Gly Val Val 1 5 10
12610PRTArtificialAn artificially synthesized peptide sequence
126Thr Leu Leu Ser Ile Phe Val Arg Val Met 1 5 10
12710PRTArtificialAn artificially synthesized peptide sequence
127Ser Ile Phe Val Arg Val Met Glu Ser Leu 1 5 10
12810PRTArtificialAn artificially synthesized peptide sequence
128Leu Thr Leu Leu Ser Ile Phe Val Arg Val 1 5 10
1299PRTArtificialAn artificially synthesized peptide sequence
129Leu Tyr Phe Asp Asp Glu Tyr Asn Ile 1 5 1309PRTArtificialAn
artifically synthesized peptide sequence 130Leu Phe Glu Arg Gly Glu
Arg Arg Leu 1 5 1319PRTArtificialAn artificially synthesized
peptide sequence 131Arg Ala Leu Gly Ala Ala Cys Leu Leu 1 5
1329PRTArtificialAn artificially synthesized peptide sequence
132Glu Tyr Asn Ile Val Lys Arg Asp Val 1 5 13310PRTArtificialAn
artificially synthesized peptide sequence 133Leu Tyr Leu Lys Leu
Leu Pro Tyr Val Leu 1 5 10 13410PRTArtificialAn artificially
synthesized peptide sequence 134Leu Phe Val Val Gln Ala Ser Leu Trp
Leu 1 5 10 13510PRTArtificialAn artificially synthesized peptide
sequence 135Arg Gln Gln Phe Phe Ile Asp Phe Arg Leu 1 5 10
13610PRTArtificialDFLEAVKRHI 136Asp Phe Leu Glu Ala Val Lys Arg His
Ile 1 5 10 13710PRTArtificialAn artificially synthesized peptide
sequence 137Arg Ala Leu Gly Ala Ala Cys Leu Leu Leu 1 5 10
13810PRTArtificialAn artificially synthesized peptide sequence
138Arg Pro Phe Val Val Val Gln Ala Arg Leu 1 5 10
13910PRTArtificialAn artificially synthesized peptide sequence
139Ala Tyr Leu Ala Gly Val Pro Gly Ser Ala 1 5 10
1409PRTArtificialAn artificially synthesized peptide sequence
140Ser Leu Trp Leu Tyr Leu Lys Leu Leu 1 5 1419PRTArtificialAn
artificially synthesized peptide sequence 141Leu Leu Leu Leu Ala
Ala Gly Trp Leu 1 5 1429PRTArtificialAn artificially synthesized
peptide sequence 142Asn Leu Phe Val Val Gln Ala Ser Leu 1 5
1439PRTArtificialAn artificially synthesized peptide sequence
143Asn Met Val Glu Lys Arg Val Asp Leu 1 5 1449PRTArtificialAn
artificially synthesized peptide sequence 144Phe Val Val Gln Ala
Ser Leu Trp Leu 1 5 1459PRTArtificialAn artificially synthesized
peptide sequence 145Gln Gln Phe Phe Ile Asp Phe Arg Leu 1 5
1469PRTArtificialAn artificially synthesized peptide sequence
146Arg Leu Gly Asp Ser Arg His Arg Ile 1 5 1479PRTArtificialAn
artificially synthesized peptide sequence 147Val Gln Ala Ser Leu
Trp Leu Tyr Leu 1 5 1489PRTArtificialAn artificially synthesized
peptide sequence 148Glu Leu Ala Val Val Pro Val Phe Val 1 5
1499PRTArtificialAn artificially synthesized peptide sequence
149Arg Leu Ile Gly Trp Asn Asp Trp Ile 1 5 1509PRTArtificialAn
artificially synthesized peptide sequence 150Arg Val Ser Glu Ile
Ile Ser Phe Ala 1 5 1519PRTArtificialAn artificially synthesized
peptide sequence 151Gly Leu Ala Ser Ser Arg Val Arg Leu 1 5
1529PRTArtificialAn artificially synthesized peptide sequence
152Ala Leu Gly Ala Ala Cys Leu Leu Leu 1 5 1539PRTArtificialAn
artificially synthesized peptide sequence 153Val Gln Cys Asp Ser
Cys Gln Glu Leu 1 5 15410PRTArtificialAn artificially synthesized
peptide sequence 154Trp Leu Tyr Leu Lys Leu Leu Pro Tyr Val 1 5 10
15510PRTArtificialAn artificially synthesized peptide sequence
155Asn Leu Cys Cys Arg Gln Gln Phe Phe Ile 1 5 10
15610PRTArtificialAn artificially synthesized peptide sequence
156Ala Leu Phe Glu Arg Gly Glu Arg Arg Leu 1 5 10
15710PRTArtificialAn artificially synthesized peptide sequence
157Met Leu Tyr Phe Asp Asp Glu Tyr Asn Ile 1 5 10
15810PRTArtificialAn artificially synthesized peptide sequence
158Cys Leu Leu Leu Leu Ala Ala Gly Trp Leu 1 5 10
15910PRTArtificialAn artificially synthesized peptide sequence
159Ala Leu Gly Ala Ala Cys Leu Leu Leu Leu 1 5 10
16010PRTArtificialAn artificially synthesized peptide sequence
160Arg Leu Ile Gly Trp Asn Asp Trp Ile Ile 1 5 10
16110PRTArtificialAn artificially synthesized peptide sequence
161Val Val Gln Ala Ser Leu Trp Leu Tyr Leu 1 5 10
16210PRTArtificialAn artificially synthesized peptide sequence
162Gln Glu Leu Ala Val Val Pro Val Phe Val 1 5 10
16310PRTArtificialAn artificially synthesized peptide sequence
163Phe Ile Ser Asn Glu Gly Asn Gln Asn Leu 1 5 10
16410PRTArtificialAn artificially synthesized peptide sequence
164Arg Gln Gln Phe Phe Ile Asp Phe Arg Leu 1 5 10
16510PRTArtificialAn artificially synthesized peptide sequence
165Gly Leu Asn Pro Gly Thr Val Asn Ser Cys 1 5 10
16610PRTArtificialAn artificially synthesized peptide sequence
166Arg Leu Gln Met Arg Gly Arg Pro Asn Ile 1 5 10
16710PRTArtificialAn artificially synthesized peptide sequence
167Arg Val Asp Gly Asp Phe Leu Glu Ala Val 1 5 10
1689PRTArtificialAn artificially synthesized peptide sequence
168Ile Tyr Asn Glu Leu Leu Tyr Asp Leu 1 5
1699PRTArtificialMYEEKLNIL 169Met Tyr Glu Glu Lys Leu Asn Ile Leu 1
5 1709PRTArtificialAn artificially synthesized peptide sequence
170Val Tyr Leu Arg Val Arg Pro Leu Leu 1 5 1719PRTArtificialAn
artificially synthesized peptide sequence 171Lys Phe Ser Ala Ile
Ala Ser Gln Leu 1 5 1729PRTArtificialAn artificially synthesized
peptide sequence 172Ser Phe Phe Glu Ile Tyr Asn Glu Leu 1 5
1739PRTArtificialAn artificially synthesized peptide sequence
173Ile Phe Asn Ser Leu Gln Gly Gln Leu 1 5 1749PRTArtificialAn
artificially synthesized peptide sequence 174Phe Phe Glu Ile Tyr
Asn Glu Leu Leu 1 5 1759PRTArtificialAn artificially synthesized
peptide sequence 175Met Phe Glu Ser Thr Ala Ala Asp Leu 1 5
1769PRTArtificialAn artificially synthesized peptide sequence
176Ser Phe Asp Ser Gly Ile Ala Gly Leu 1 5 1779PRTArtificialAn
artificially synthesized peptide sequence 177Arg Phe Ser Ile Trp
Ile Ser Phe Phe 1 5 1789PRTArtificialAn artificially synthesized
peptide sequence 178Ile Phe Ser Ile Arg Ile Leu His Leu 1 5
1799PRTArtificialAn artificially synthesized peptide sequence
179Lys Ile Glu Glu Leu Glu Ala Leu Leu 1 5 1809PRTArtificialAn
artificially synthesized peptide sequence 180Lys Leu Asn Ile Leu
Lys Glu Ser Leu 1 5 1819PRTArtificialAn artificially synthesized
peptide sequence 181Lys Leu Gln Gln Cys Lys Ala Glu Leu 1 5
1829PRTArtificialAn artificially synthesized peptide sequence
182Phe Thr Ile Asp Val Asp Lys Lys Leu 1 5 1839PRTArtificialAn
artificially synthesized peptide sequence 183Gln Leu Gln Glu Val
Lys Ala Lys Leu 1 5 18410PRTArtificialAn artificially synthesized
peptide sequence 184Ile Tyr Asn Glu Leu Leu Tyr Asp Leu Leu 1 5 10
18510PRTArtificialAn artificially synthesized peptide sequence
185Arg Ser Leu Ala Leu Ile Phe Asn Ser Leu 1 5 10
18610PRTArtificialAn artificially synthesized peptide sequence
186Ser Phe Phe Glu Ile Tyr Asn Glu Leu Leu 1 5 10
18710PRTArtificialAn artificially synthesized peptide sequence
187Arg Leu Leu Arg Thr Glu Leu Gln Lys Leu 1 5 10
18810PRTArtificialAn artificially synthesized peptide sequence
188Lys Asn Ile Arg Leu Leu Arg Thr Glu Leu 1 5 10
18910PRTArtificialAn artificially synthesized peptide sequence
189Arg Gln Glu Glu Met Lys Lys Leu Ser Leu 1 5 10
19010PRTArtificialAn artificially synthesized peptide sequence
190Arg Val Arg Pro Leu Leu Pro Ser Glu Leu 1 5 10
19110PRTArtificialAn artificially synthesized peptide sequence
191Arg Ile Leu Arg Ser Arg Arg Ser Pro Leu 1 5 10
19210PRTArtificialAn artificially synthesized peptide sequence
192Arg Ile Glu Asn Val Glu Thr Leu Val Leu 1 5 10
19310PRTArtificialAn artificially synthesized peptide sequence
193Lys Asn Gln Ser Phe Ala Ser Thr His Leu 1 5 10
19410PRTArtificialAn artificially synthesized peptide sequence
194Lys Val Tyr Leu Arg Val Arg Pro Leu Leu 1 5 10
19510PRTArtificialAn artificially synthesized peptide sequence
195Asp Ser Met Glu Lys Val Lys Val Tyr Leu 1 5 10
1969PRTArtificialAn artificially synthesized peptide sequence
196Lys Tyr Gln Ala Tyr Met Ser Asn Leu 1 5 1979PRTArtificialAn
artificially synthesized peptide sequence 197Val Tyr Val Pro Leu
Lys Glu Leu Leu 1 5 1989PRTArtificialAn artificially synthesized
peptide sequence 198Glu Tyr His Lys Leu Ala Arg Lys Leu 1 5
1999PRTArtificialAn artificially synthesized peptide sequence
199Ser Tyr Glu Leu Pro Asp Thr Lys Phe 1 5 2009PRTArtificialAn
artificially synthesized peptide sequence 200Lys Tyr Glu Lys Lys
Ala Thr Leu Ile 1 5 2019PRTArtificialAn artificially synthesized
peptide sequence 201Lys Tyr Ala Arg Gly Lys Glu Ala Ile 1 5
2029PRTArtificialAn artificially synthesized peptide sequence
202Asp Phe Leu Lys Ile Phe Thr Phe Leu 1 5 2039PRTArtificialAn
artificially synthesized peptide sequence 203Gly Phe Leu Cys Pro
Ser Tyr Glu Leu 1 5 2049PRTArtificialAn artificially synthesized
peptide sequence 204Leu Phe Asn Val Asp Ala Phe Lys Leu 1 5
2059PRTArtificialAn artificially synthesized peptide sequence
205Ser Phe Asp Glu Met Asn Ala Glu Leu 1 5 2069PRTArtificialAn
artificially synthesized peptide sequence 206Ile Phe Thr Phe Leu
Tyr Gly Phe Leu 1 5 2079PRTArtificialAn artificially synthesized
peptide sequence 207Lys Phe Glu Glu Glu Val Pro Arg Ile 1 5
2089PRTArtificialAn artificially synthesized peptide sequence
208Arg Ile Asn His Glu Arg Asn Glu Leu 1 5 2099PRTArtificialAn
artificially synthesized peptide sequence 209Ser Phe
Met Ser Gly Ala Asp Ser Phe 1 5 2109PRTArtificialAn artificially
synthesized peptide sequence 210Ile Phe Lys Asp Leu Gly Tyr Pro Phe
1 5 2119PRTArtificialAn artificially synthesized peptide sequence
211Glu Tyr Gln Leu Val Val Gln Thr Thr 1 5 2129PRTArtificialAn
artificially synthesized peptide sequence 212Lys Ala Leu Asn Lys
Lys Met Gly Leu 1 5 2139PRTArtificialAn artificially synthesized
peptide sequence 213Glu Val Pro Arg Ile Phe Lys Asp Leu 1 5
21410PRTArtificialAn artificially synthesized peptide sequence
214Lys Tyr Arg Ala Gln Val Tyr Val Pro Leu 1 5 10
21510PRTArtificialAn artificially synthesized peptide sequence
215Glu Tyr Glu Glu Cys Met Ser Glu Asp Leu 1 5 10
21610PRTArtificialAn artificially synthesized peptide sequence
216Lys Tyr Ser Val Ala Asp Ile Glu Arg Ile 1 5 10
21710PRTArtificialAn artificially synthesized peptide sequence
217Asp Tyr Thr Ile Lys Cys Tyr Glu Ser Phe 1 5 10
21810PRTArtificialAn artificially synthesized peptide sequence
218Lys Phe Glu Glu Glu Val Pro Arg Ile Phe 1 5 10
21910PRTArtificialAn artificially synthesized peptide sequence
219Ala Phe Ile Gln Gln Cys Ile Arg Gln Leu 1 5 10
22010PRTArtificialAn artificially synthesized peptide sequence
220Arg Ser Gln Asp Val Asn Lys Gln Gly Leu 1 5 10
22110PRTArtificialAn artificially synthesized peptide sequence
221Arg Thr Leu Lys Glu Glu Val Gln Lys Leu 1 5 10
22210PRTArtificialAn artificially synthesized peptide sequence
222Arg Gly Lys Glu Ala Ile Glu Thr Gln Leu 1 5 10
22310PRTArtificialAn artificially synthesized peptide sequence
223Arg Ala Leu Asn Glu Gln Ile Ala Arg Leu 1 5 10
22410PRTArtificialAn artificially synthesized peptide sequence
224Glu Tyr Gln Leu Val Val Gln Thr Thr Thr 1 5 10
22510PRTArtificialAn artificially synthesized peptide sequence
225Glu Thr Glu Glu Glu Ile Asn Lys Ala Leu 1 5 10
22610PRTArtificialAn artificially synthesized peptide sequence
226Leu Leu Glu Ser Thr Val Asn Gln Gly Leu 1 5 10
2279PRTArtificialAn artificially synthesized peptide sequence
227Tyr Met Ser Cys Phe Arg Thr Pro Val 1 5 2289PRTArtificialAn
artificially synthesized peptide sequence 228Lys Gln Ile Tyr Ala
Ile Lys Tyr Val 1 5 2299PRTArtificialAn artificially synthesized
peptide sequence 229Asn Met Leu Glu Ala Val His Thr Ile 1 5
2309PRTArtificialAn artificially synthesized peptide sequence
230Leu Leu Asn Ser Pro Asp Cys Asp Val 1 5 2319PRTArtificialAn
artificially synthesized peptide sequence 231Ile Leu Ala Thr Pro
Leu Gln Asn Leu 1 5 2329PRTArtificialAn artificially synthesized
peptide sequence 232Tyr Val Leu Gly Gln Leu Val Gly Leu 1 5
2339PRTArtificialAn artificially synthesized peptide sequence
233Ser Leu Gly Cys Ile Leu Tyr Tyr Met 1 5 2349PRTArtificialAn
artificially synthesized peptide sequence 234Gln Met Gln Pro Asp
Thr Thr Ser Val 1 5 2359PRTArtificialAn artificially synthesized
peptide sequence 235Gly Thr Thr Glu Glu Met Lys Tyr Val 1 5
2369PRTArtificialAn artificially synthesized peptide sequence
236Leu Ile Val Asp Gly Met Leu Lys Leu 1 5 2379PRTArtificialAn
artificially synthesized peptide sequence 237Ser Leu Leu Ala Lys
Leu Glu Glu Thr 1 5 2389PRTArtificialAn artificially synthesized
peptide sequence 238Leu Phe Glu Arg Gly Glu Arg Arg Leu 1 5
2399PRTArtificialAn artificially synthesized peptide sequence
239Leu Leu Ala His Pro Tyr Val Gln Ile 1 5 2409PRTArtificialAn
artificially synthesized peptide sequence 240Lys Leu Ile Gly Arg
Tyr Ser Gln Ala 1 5 2419PRTArtificialAn artificially synthesized
peptide sequence 241Asn Leu Asn Leu Gln Lys Lys Gln Leu 1 5
2429PRTArtificialAn artificially synthesized peptide sequence
242Met Gln Pro Asp Thr Thr Ser Val Val 1 5 2439PRTArtificialAn
artificially synthesized peptide sequence 243Lys Leu Gln Gln His
Ser Asp Lys Ile 1 5 2449PRTArtificialAn artificially synthesized
peptide sequence 244Phe Ala Phe Val His Ile Ser Phe Ala 1 5
2459PRTArtificialAn artificially synthesized peptide sequence
245Cys Glu Leu Arg Asn Leu Lys Ser Val 1 5 2469PRTArtificialAn
artificially synthesized peptide sequence 246Ser Ile Leu Lys Ala
Ala Lys Thr Leu 1 5 24710PRTArtificialAn artificially synthesized
peptide sequence 247Leu Leu Leu Lys Leu Glu Lys Asn Ser Val 1 5 10
24810PRTArtificialAn artificially synthesized peptide sequence
248Asn Leu Leu Asn Ser Pro Asp Cys Asp Val 1 5 10
24910PRTArtificialAn artificially synthesized peptide sequence
249Phe Leu Ile Val Asp Gly Met Leu Lys Leu 1 5 10
25010PRTArtificialAn artificially synthesized peptide sequence
250Thr Thr Phe Glu Gln Pro Val Phe Ser Val 1 5 10
25110PRTArtificialAn artificially synthesized peptide sequence
251Val Leu Asn Glu Lys Lys Gln Ile Tyr Ala 1 5 10
25210PRTArtificialAn artificially synthesized peptide sequence
252Gly Met Leu Lys Leu Ile Asp Phe Gly Ile 1 5 10
25310PRTArtificialAn artificially synthesized peptide sequence
253Leu Leu Ser Glu Glu Glu Lys Lys Asn Leu 1 5 10
25410PRTArtificialAn artificially synthesized peptide sequence
254Tyr Met Ser Cys Phe Arg Thr Pro Val Val 1 5 10
25510PRTArtificialAn artificially synthesized peptide sequence
255Met Met Ala Asn Asn Pro Glu Asp Trp Leu 1 5 10
25610PRTArtificialAn artificially synthesized peptide sequence
256Met Val Met Glu Cys Gly Asn Ile Asp Leu 1 5 10
25710PRTArtificialAn artificially synthesized peptide sequence
257Tyr Met Pro Pro Glu Ala Ile Lys Asp Met 1 5 10
25810PRTArtificialAn artificially synthesized peptide sequence
258Lys Leu Ile Gly Arg Tyr Ser Gln Ala Ile 1 5 10
25910PRTArtificialAn artificially synthesized peptide sequence
259Asn Gln Met Gln Pro Asp Thr Thr Ser Val 1 5 10
26010PRTArtificialAn artificially synthesized peptide sequence
260Gln Ile Leu Ala Thr Pro Leu Gln Asn Leu 1 5 10
26110PRTArtificialAn artificially synthesized peptide sequence
261Leu Ile Val Asp Gly Met Leu Lys Leu Ile 1 5 10
26210PRTArtificialAn artificially synthesized peptide sequence
262Asn Leu Asn Leu Gln Lys Lys Gln Leu Leu 1 5 10
26310PRTArtificialAn artificially synthesized peptide sequence
263Gln Met Gln Pro Asp Thr Thr Ser Val Val 1 5 10
26410PRTArtificialAn artificially synthesized peptide sequence
264Lys Gly Thr Thr Glu Glu Met Lys Tyr Val 1 5 10
26510PRTArtificialAn artificially synthesized peptide sequenc
265Leu Thr Ile Asp Ser Ile Met Asn Lys Val 1 5 10
26610PRTArtificialAn artificially synthesized peptide sequence
266Lys Leu Gln Gln His Ser Asp Lys Ile Ile 1 5 10
2679PRTArtificialAn artificially synthesized peptide sequence
267Lys Ile Phe Pro Arg Phe Phe Met Val 1 5 2689PRTArtificialAn
artificially synthesized peptide sequence 268Gly Leu Trp Leu Ala
Ile Leu Leu Leu 1 5 2699PRTArtificialAn artificially synthesized
peptide sequence 269Leu Leu Val Val Ala Leu Pro Arg Val 1 5
2709PRTArtificialAn artificially synthesized peptide sequence
270Ala Leu Leu Ala Leu Leu Leu Val Val 1 5 2719PRTArtificialAn
artificially synthesized peptide sequence 271Trp Leu Ala Ile Leu
Leu Leu Leu Ala 1 5 2729PRTArtificialAn artificially synthesized
peptide sequence 272Leu Leu Ala Ser Ile Ala Ala Gly Leu 1 5
2739PRTArtificialAn artificially synthesized peptide sequence
273Leu Leu Leu Leu Ala Ser Ile Ala Ala 1 5 2749PRTArtificialAn
artificially synthesized peptide sequence 274Phe Met Val Ala Lys
Gln Cys Ser Ala 1 5 2759PRTArtificialAn artificially synthesized
peptide sequence 275Thr Met Ala Leu Leu Ala Leu Leu Leu 1 5
2769PRTArtificialAn artificially synthesized peptide sequence
276Met Ala Leu Leu Ala Leu Leu Leu Val 1 5 2779PRTArtificialAn
artificially synthesized peptide sequence 277Ala Ile Leu Leu Leu
Leu Ala Ser Ile 1 5 2789PRTArtificialAn artificially synthesized
peptide sequence 278Ala Leu Pro Arg Val Trp Thr Asp Ala 1 5
2799PRTArtificialAn artificially synthesized peptide sequence
279Ser Met Gly Glu Ser Cys Gly Gly Leu 1 5 2809PRTArtificialAn
artificially synthesized peptide sequence 280Leu Leu Ala Leu Leu
Leu Val Val Ala 1 5 2819PRTArtificialAn artificially synthesized
peptide sequence 281Val Val Ala Leu Pro Arg Val Trp Thr 1 5
2829PRTArtificialAn artificially synthesized peptide sequence
282Arg Val Trp Thr Asp Ala Asn Leu Thr 1 5 2839PRTArtificialAn
artificially synthesized peptide sequence 283Phe Leu Leu Glu Glu
Pro Met Pro Phe 1 5 2849PRTArtificialAn artificially synthesized
peptide sequence 284Leu Ala Leu Leu Leu Val Val Ala Leu 1 5
2859PRTArtificialAn artificially synthesized peptide sequence
285Gly Thr Met Ala Leu Leu Ala Leu Leu 1 5 28610PRTArtificialAn
artificially synthesized peptide sequence 286Leu Leu Leu Val Val
Ala Leu Pro Arg Val 1 5 10 28710PRTArtificialAn artificially
synthesized peptide sequence 287Gly Leu Trp Leu Ala Ile Leu Leu Leu
Leu 1 5 10 28810PRTArtificialAn artificially synthesized peptide
sequence 288Leu Leu Leu Ala Ser Ile Ala Ala Gly Leu 1 5 10
28910PRTArtificialAn artificially synthesized peptide sequence
289Thr Met Ala Leu Leu Ala Leu Leu Leu Val 1 5 10
29010PRTArtificialAn artificially synthesized peptide sequence
290Leu Leu Ala Leu Leu Leu Val Val Ala Leu 1 5 10
29110PRTArtificialAn artificially synthesized peptide sequence
291Phe Leu Leu Glu Glu Pro Met Pro Phe Phe 1 5 10
29210PRTArtificialAn artificially synthesized peptide sequence
292Ile Leu Leu Leu Leu Ala Ser Ile Ala Ala 1 5 10
29310PRTArtificialAn artificially synthesized peptide sequence
293Lys Ile Phe Pro Arg Phe Phe Met Val Ala 1 5 10
29410PRTArtificialAn artificially synthesized peptide sequence
294Ala Leu Leu Ala Leu Leu Leu Val Val Ala 1 5 10
29510PRTArtificialAn artificially synthesized peptide sequence
295Leu Val Val Ala Leu Pro Arg Val Trp Thr 1 5 10
29610PRTArtificialAn artificially synthesized peptide sequence
296Met Ala Leu Leu Ala Leu Leu Leu Val Val 1 5 10
29710PRTArtificialAn artificially synthesized peptide sequence
297Arg Leu Gln Arg Pro Arg Gln Ala Pro Ala 1 5 10
29810PRTArtificialAn artificially synthesized peptide sequence
298Cys Gln Asn Pro Arg Arg Cys Lys Trp Thr 1 5 10
29910PRTArtificialAn artificially synthesized peptide sequence
299Arg Val Trp Thr Asp Ala Asn Leu Thr Ala 1 5 10
30010PRTArtificialAn artificially synthesized peptide sequence
300Trp Ala Pro Leu Gly Thr Met Ala Leu Leu 1 5 10
30110PRTArtificialAn artificially synthesized peptide sequence
301Thr Glu Pro Tyr Cys Val Ile Ala Ala Val 1 5 10
30210PRTArtificialAn artificially synthesized peptide sequence
302Leu Glu Glu Pro Met Pro Phe Phe Tyr Leu 1 5 10
30310PRTArtificialAn artificially synthesized peptide sequence
303Leu Glu Gly Pro Pro Ile Asn Ser Ser Val 1 5 10
30410PRTArtificialAn artificially synthesized peptide sequence
304Tyr Leu Lys Cys Cys Lys Ile Arg Tyr Cys 1 5 10
30510PRTArtificialAn artificially synthesized peptide sequence
305Val Lys Ile Phe Pro Arg Phe Phe Met Val 1 5 10
3069PRTArtificialAn artificially synthesized peptide sequence
306Lys Ile Phe Pro Ser Lys Arg Ile Leu 1 5 3079PRTArtificialAn
artificially synthesized peptide sequence 307Arg Gly Ser Val Leu
Glu Gly Val Leu 1 5 3089PRTArtificialAn artificially synthesized
peptide sequence 308Phe Leu Leu Leu Val Leu Leu Leu Leu 1 5
3099PRTArtificialAn artificially synthesized peptide sequence
309Ile Gly Asn Phe Ile Ile Glu Asn Leu 1 5 3109PRTArtificialAn
artificially synthesized peptide sequence 310Thr Ala Val Ala Val
Val Glu Ile Leu 1 5 3119PRTArtificialAn artificially synthesized
peptide sequence 311Asn Gln Ser Pro Val Arg Gln Val Leu 1 5
3129PRTArtificialAn artificially synthesized peptide sequence
312Lys Gln Asp Thr Tyr Asp Val His Leu 1 5 3139PRTArtificialAn
artificially synthesized peptide sequence 313Asp Tyr Glu Gly Ser
Gly Ser Asp Ala 1 5 3149PRTArtificialAn artificially synthesized
peptide sequence 314Gly Trp Leu Leu Leu Asn Lys Pro Leu 1 5
3159PRTArtificialAn artificially synthesized peptide sequence
315Ile Leu Pro Val Leu Gly Ala Val Leu 1 5 3169PRTArtificialAn
artificially synthesized peptide sequence 316Thr Ala Pro Pro Tyr
Asp Thr Leu Leu 1 5 3179PRTArtificialAn artificially synthesized
peptide sequence 317Val Val Leu Ser Leu Lys Lys Phe Leu 1 5
3189PRTArtificialAn artificially synthesized peptide sequence
318Ala Leu Leu Phe Leu Leu Leu Val Leu 1 5 3199PRTArtificialAn
artificially synthesized peptide sequence 319Val Thr Asn Glu Ala
Pro Phe Val Leu 1 5 3209PRTArtificialAn artificially synthesized
peptide sequence 320Ala Val Leu Ala Leu Leu Phe Leu Leu 1 5
3219PRTArtificialAn artificially synthesized peptide sequence
321Asp Thr Tyr Asp Val His Leu Ser Leu 1 5 3229PRTArtificialAn
artificially synthesized peptide sequence 322Gly Pro Leu Ala Ser
Leu Leu Leu Leu 1 5 3239PRTArtificialAn artificially synthesized
peptide sequence 323Val Leu Asn Ile Thr Asp Lys Asp Leu 1 5
3249PRTArtificialAn artificially synthesized peptide sequence
324Ala Val Glu Lys Glu Thr Gly Trp Leu 1 5 3259PRTArtificialAn
artificially synthesized peptide sequence 325Asn Asn Ile Tyr Glu
Val Met Val Leu 1 5 3269PRTArtificialAn artificially synthesized
peptide sequence 326Leu Leu Leu Leu Gln Val Cys Trp Leu 1 5
3279PRTArtificialAn artificially synthesized peptide sequence
327Gly Cys Pro Gly Gln Glu Pro Ala Leu 1 5 3289PRTArtificialAn
artificially synthesized peptide sequence 328Glu Tyr Thr Leu Thr
Ile Gln Ala Thr 1 5 3299PRTArtificialAn artificially synthesized
peptide sequence 329Glu Thr Val Gln Glu Arg Arg Ser Leu 1 5
3309PRTArtificialAn artificially synthesized peptide sequence
330Ser Tyr Arg Ile Leu Arg Asp Pro Ala 1 5 3319PRTArtificialAn
artificially synthesized peptide sequence 331Gly Gln Val Thr Ala
Val Gly Thr Leu 1 5 3329PRTArtificialAn artificially synthesized
peptide sequence 332Gly Ala Val Leu Ala Leu Leu Phe Leu 1 5
3339PRTArtificialAn artificially synthesized peptide sequence
333Gly Ile Leu Thr Thr Arg Lys Gly Leu 1 5 3349PRTArtificialAn
artificially synthesized peptide sequence 334His Pro Glu Ser Asn
Gln Gly Ile Leu 1 5
3359PRTArtificialAn artificially synthesized peptide sequence
335Val Leu Ala Leu Leu Phe Leu Leu Leu 1 5 3369PRTArtificialAn
artificially synthesized peptide sequence 336Glu Gly Asp Thr Val
Val Leu Ser Leu 1 5 3379PRTArtificialAn artificially synthesized
peptide sequence 337Thr Ile Ser Val Ile Ser Ser Gly Leu 1 5
3389PRTArtificialAn artificially synthesized peptide sequence
338Val Leu Gly Ala Val Leu Ala Leu Leu 1 5 3399PRTArtificialAn
artificially synthesized peptide sequence 339Glu Trp Gly Ser Arg
Phe Lys Lys Leu 1 5 3409PRTArtificialAn artificially synthesized
peptide sequence 340Lys Val Val Glu Val Gln Glu Gly Ile 1 5
3419PRTArtificialAn artificially synthesized peptide sequence
341Thr Tyr Asp Val His Leu Ser Leu Ser 1 5 3429PRTArtificialAn
artificially synthesized peptide sequence 342Phe Tyr Ser Ile Thr
Gly Pro Gly Ala 1 5 3439PRTArtificialAn artificially synthesized
peptide sequence 343Ile Tyr Thr Tyr Asn Gly Val Val Ala 1 5
34410PRTArtificialAn artificially synthesized peptide sequence
344Phe Ile Leu Pro Val Leu Gly Ala Val Leu 1 5 10
34510PRTArtificialAn artificially synthesized peptide sequence
345Ala Val Leu Ala Leu Leu Phe Leu Leu Leu 1 5 10
34610PRTArtificialAn artificially synthesized peptide sequence
346Gly Thr Ile Ser Val Ile Ser Ser Gly Leu 1 5 10
34710PRTArtificialAn artificially synthesized peptide sequence
347Asp Tyr Glu Gly Ser Gly Ser Asp Ala Ala 1 5 10
34810PRTArtificialAn artificially synthesized peptide sequence
348Thr Val Val Leu Ser Leu Lys Lys Phe Leu 1 5 10
34910PRTArtificialAn artificially synthesized peptide sequence
349Phe Ala Val Glu Lys Glu Thr Gly Trp Leu 1 5 10
35010PRTArtificialAn artificially synthesized peptide sequence
350Ala Leu Leu Phe Leu Leu Leu Val Leu Leu 1 5 10
35110PRTArtificialAn artificially synthesized peptide sequence
351Ser Gln Glu Pro Lys Asp Pro His Asp Leu 1 5 10
35210PRTArtificialAn artificially synthesized peptide sequence
352Leu Ala Leu Leu Phe Leu Leu Leu Val Leu 1 5 10
35310PRTArtificialAn artificially synthesized peptide sequence
353Gly Ala Glu Gln Glu Pro Gly Gln Ala Leu 1 5 10
35410PRTArtificialAn artificially synthesized peptide sequence
354Gly Ala Val Leu Ala Leu Leu Phe Leu Leu 1 5 10
35510PRTArtificialAn artificially synthesized peptide sequence
355Val Asn Glu Glu Gly Asp Thr Val Val Leu 1 5 10
35610PRTArtificialAn artificially synthesized peptide sequence
356Asn Ala Val Gly His Glu Val Gln Arg Leu 1 5 10
35710PRTArtificialAn artificially synthesized peptide sequence
357Thr Asn Glu Ala Pro Phe Val Leu Lys Leu 1 5 10
35810PRTArtificialAn artificially synthesized peptide sequence
358Glu Asn Gln Lys Ile Ser Tyr Arg Ile Leu 1 5 10
35910PRTArtificialAn artificially synthesized peptide sequence
359Ser Leu Leu Leu Leu Gln Val Cys Trp Leu 1 5 10
36010PRTArtificialAn artificially synthesized peptide sequence
360Gly Leu Glu Ala Arg Pro Glu Val Val Leu 1 5 10
36110PRTArtificialAn artificially synthesized peptide sequence
361Glu Val Gln Arg Leu Thr Val Thr Asp Leu 1 5 10
36210PRTArtificialAn artificially synthesized peptide sequence
362Gly Leu Pro Arg Gly Pro Leu Ala Ser Leu 1 5 10
36310PRTArtificialAn artificially synthesized peptide sequence
363Leu Pro Val Leu Gly Ala Val Leu Ala Leu 1 5 10
36410PRTArtificialAn artificially synthesized peptide sequence
364Gln Val Leu Asn Ile Thr Asp Lys Asp Leu 1 5 10
36510PRTArtificialAn artificially synthesized peptide sequence
365Ala Val Glu Lys Glu Thr Gly Trp Leu Leu 1 5 10
36610PRTArtificialAn artificially synthesized peptide sequence
366Ser Gly Gln Val Thr Ala Val Gly Thr Leu 1 5 10
36710PRTArtificialAn artificially synthesized peptide sequence
367Gln Gly Ile Leu Thr Thr Arg Lys Gly Leu 1 5 10
36810PRTArtificialAn artificially synthesized peptide sequence
368Ser Pro Pro Thr Thr Gly Thr Gly Thr Leu 1 5 10
36910PRTArtificialAn artificially synthesized peptide sequence
369Asn Ser Pro Ala Trp Arg Ala Thr Tyr Leu 1 5 10
37010PRTArtificialAn artificially synthesized peptide sequence
370Gly Pro Phe Pro Gln Arg Leu Asn Gln Leu 1 5 10
37110PRTArtificialAn artificially synthesized peptide sequence
371Glu Ile Gly Asn Phe Ile Ile Glu Asn Leu 1 5 10
37210PRTArtificialAn artificially synthesized peptide sequence
372Thr Thr Ala Val Ala Val Val Glu Ile Leu 1 5 10
37310PRTArtificialAn artificially synthesized peptide sequence
373Ile Tyr Thr Tyr Asn Gly Val Val Ala Tyr 1 5 10
37410PRTArtificialAn artificially synthesized peptide sequence
374Asp Tyr Asp Tyr Leu Asn Glu Trp Gly Ser 1 5 10
3759PRTArtificialAn artificially synthesized peptide sequence
375Ala Leu Phe Ser Cys Leu Phe Gly Ile 1 5 3769PRTArtificialAn
artificially synthesized peptide sequence 376Gly Leu Asn Pro Leu
Thr Ser Tyr Val 1 5 3779PRTArtificialAn artificially synthesized
peptide sequence 377Cys Leu Phe Gly Ile Cys Asp Ala Val 1 5
3789PRTArtificialAn artificially synthesized peptide sequence
378Gln Met His Gly Arg Met Val Pro Val 1 5 3799PRTArtificialAn
artificially synthesized peptide sequence 379Lys Leu Asn Thr Glu
Ile Arg Asp Val 1 5 3809PRTArtificialAn artificially synthesized
peptide sequence 380Trp Leu Val Pro Ile Gly Asn Cys Leu 1 5
3819PRTArtificialAn artificially synthesized peptide sequence
381Lys Leu Ile Arg Asn Pro Asn Ser Leu 1 5 3829PRTArtificialAn
artificially synthesized peptide sequence 382Val Val Ile Leu Ile
Ala Ala Phe Val 1 5 3839PRTArtificialAn artificially synthesized
peptide sequence 383Val Met Trp Glu Val Met Ser Tyr Gly 1 5
3849PRTArtificialAn artificially synthesized peptide sequence
384Gly Ile Gly Ser Gly Met Lys Tyr Leu 1 5 3859PRTArtificialAn
artificially synthesized peptide sequence 385Asn Ile Leu Val Asn
Ser Asn Leu Val 1 5 3869PRTArtificialAn artificially synthesized
peptide sequence 386Thr Thr Leu Glu Ala Val Val His Val 1 5
3879PRTArtificialAn artificially synthesized peptide sequence
387Tyr Leu Leu Gly Val Val Leu Thr Leu 1 5 3889PRTArtificialAn
artificially synthesized peptide sequence 388Leu Tyr Leu Leu Gly
Val Val Leu Thr 1 5 3899PRTArtificialAn artificially synthesized
peptide sequence 389Val Met Glu Ser Leu Glu Gly Leu Leu 1 5
3909PRTArtificialAn artificially synthesized peptide sequence
390Leu Leu Gly Val Val Leu Thr Leu Leu 1 5 39110PRTArtificialAn
artificially synthesized peptide sequence 391Tyr Leu Leu Gly Val
Val Leu Thr Leu Leu 1 5 10 39210PRTArtificialAn artificially
synthesized peptide sequence 392Leu Asn Leu Tyr Leu Leu Gly Val Val
Leu 1 5 10 39310PRTArtificialAn artificially synthesized peptide
sequence 393Leu Ala Asn Thr Glu Pro Thr Lys Gly Leu 1 5 10
39410PRTArtificialAn artificially synthesized peptide sequence
394Ser Ile Phe Val Arg Val Met Glu Ser Leu 1 5 10
3959PRTArtificialAn artificially synthesized peptide sequence
395Leu Tyr Leu Lys Leu Leu Pro Tyr Val 1 5 3969PRTArtificialAn
artificially synthesized peptide sequence 396Ile Ser Asn Glu Gly
Asn Gln Asn Leu 1 5 3979PRTArtificialAn artificially synthesized
peptide sequence 397Arg Ser Gly Trp His Thr Phe Pro Leu 1 5
3989PRTArtificialAn artificially synthesized peptide sequence
398Ala Ser Leu Trp Leu Tyr Leu Lys Leu 1 5 3999PRTArtificialAn
artificially synthesized peptide sequence 399Ala Tyr Leu Ala Gly
Val Pro Gly Ser 1 5 4009PRTArtificialAn artificially synthesized
peptide sequence 400Asn Met Val Glu Lys Arg Val Asp Leu 1 5
4019PRTArtificialAn artificially synthesized peptide sequence
401Ala Met Val Thr Ala Leu Arg Lys Leu 1 5 4029PRTArtificialAn
artificially synthesized peptide sequence 402Val Gln Cys Asp Ser
Cys Gln Glu Leu 1 5 4039PRTArtificialAn artificially synthesized
peptide sequence 403Asn Ser Cys Cys Ile Pro Thr Lys Leu 1 5
4049PRTArtificialAn artificially synthesized peptide sequence
404Asn Tyr Cys Glu Gly Ser Cys Pro Ala 1 5 4059PRTArtificialAn
artificially synthesized peptide sequence 405Phe Val Val Gln Ala
Ser Leu Trp Leu 1 5 4069PRTArtificialAn artificially synthesized
peptide sequence 406Val Asn Gln Tyr Arg Met Arg Gly Leu 1 5
4079PRTArtificialAn artificially synthesized peptide sequence
407Gln Phe Phe Ile Asp Phe Arg Leu Ile 1 5 4089PRTArtificialAn
artificially synthesized peptide sequence 408Leu Leu Leu Leu Ala
Ala Gly Trp Leu 1 5 4099PRTArtificialAn artificially synthesized
peptide sequence 409Gln Gln Phe Phe Ile Asp Phe Arg Leu 1 5
4109PRTArtificialAn artificially synthesized peptide sequence
410Asn Leu Phe Val Val Gln Ala Ser Leu 1 5 4119PRTArtificialAn
artificially synthesized peptide sequence 411Tyr Tyr Gly Asn Tyr
Cys Glu Gly Ser 1 5 41210PRTArtificialAn artificially synthesized
peptide sequence 412Gln Asn Leu Phe Val Val Gln Ala Ser Leu 1 5 10
41310PRTArtificialAn artificially synthesized peptide sequence
413Asp Val Gln Cys Asp Ser Cys Gln Glu Leu 1 5 10
41410PRTArtificialAn artificially synthesized peptide sequence
414Val Val Gln Ala Ser Leu Trp Leu Tyr Leu 1 5 10
41510PRTArtificialAn artificially synthesized peptide sequence
415Leu Tyr Phe Asp Asp Glu Tyr Asn Ile Val 1 5 10
41610PRTArtificialAn artificially synthesized peptide sequence
416Phe Pro Leu Thr Glu Ala Ile Gln Ala Leu 1 5 10
41710PRTArtificialAn artificially synthesized peptide sequence
417Arg Thr Asn Leu Cys Cys Arg Gln Gln Phe 1 5 10
41810PRTArtificialAn artificially synthesized peptide sequence
418Ala Ala Met Val Thr Ala Leu Arg Lys Leu 1 5 10
41910PRTArtificialAn artificially synthesized peptide sequence
419Val Asn Ser Cys Cys Ile Pro Thr Lys Leu 1 5 10
42010PRTArtificialAn artificially synthesized peptide sequence
420Cys Leu Leu Leu Leu Ala Ala Gly Trp Leu 1 5 10
42110PRTArtificialAn artificially synthesized peptide sequence
421Val Val Asn Gln Tyr Arg Met Arg Gly Leu 1 5 10
42210PRTArtificialAn artificially synthesized peptide sequence
422Asp Gly Leu Ala Ser Ser Arg Val Arg Leu 1 5 10
42310PRTArtificialAn artificially synthesized peptide sequence
423Gly Leu Glu Cys Asp Gly Arg Thr Asn Leu 1 5 10
42410PRTArtificialAn artificially synthesized peptide sequence
424Asn Tyr Cys Glu Gly Ser Cys Pro Ala Tyr 1 5 10
42510PRTArtificialAn artificially synthesized peptide sequence
425Ala Ser Leu Trp Leu Tyr Leu Lys Leu Leu 1 5 10
42610PRTArtificialAn artificially synthesized peptide sequence
426Trp Asn Met Val Glu Lys Arg Val Asp Leu 1 5 10
42710PRTArtificialAn artificially synthesized peptide sequence
427Phe Leu Glu Ala Val Lys Arg His Ile Leu 1 5 10
42810PRTArtificialAn artificially synthesized peptide sequence
428Tyr Cys Glu Gly Ser Cys Pro Ala Tyr Leu 1 5 10
42910PRTArtificialAn artificially synthesized peptide sequence
429Ala Val Lys Arg His Ile Leu Ser Arg Leu 1 5 10
43010PRTArtificialAn artificially synthesized peptide sequence
430Gln Ala Ser Leu Trp Leu Tyr Leu Lys Leu 1 5 10
43110PRTArtificialAn artificially synthesized peptide sequence
431Gly Tyr Tyr Gly Asn Tyr Cys Glu Gly Ser 1 5 10
43210PRTArtificialAn artificially synthesized peptide sequence
432Leu Tyr Phe Phe Ile Ser Asn Glu Gly Asn 1 5 10
43310PRTArtificialAn artificially synthesized peptide sequence
433Tyr Tyr Gly Asn Tyr Cys Glu Gly Ser Cys 1 5 10
4349PRTArtificialAn artificially synthesized peptide sequence
434Ile Leu Leu Leu Leu Ala Ser Ile Ala 1 5 4353218DNAHomo
sapiensCDS(47)..(1270) 435actcggctcg cctcgcggcg ggcgccctcg
tcgccagcgg cgcacc atg gac ggg 55 Met Asp Gly 1 ctg ccc ggt cgg gcg
ctg ggg gcc gcc tgc ctt ctg ctg ctg gcg gcc 103Leu Pro Gly Arg Ala
Leu Gly Ala Ala Cys Leu Leu Leu Leu Ala Ala 5 10 15 ggc tgg ctg ggg
cct gag gcc tgg ggc tca ccc acg ccc ccg ccg acg 151Gly Trp Leu Gly
Pro Glu Ala Trp Gly Ser Pro Thr Pro Pro Pro Thr 20 25 30 35 cct gcc
gcg ccg ccg cca ccc ccg cca ccc gga tcc ccg ggt ggc tcg 199Pro Ala
Ala Pro Pro Pro Pro Pro Pro Pro Gly Ser Pro Gly Gly Ser 40 45 50
cag gac acc tgt acg tcg tgc ggc ggc ttc cgg cgg cca gag gag ctc
247Gln Asp Thr Cys Thr Ser Cys Gly Gly Phe Arg Arg Pro Glu Glu Leu
55 60 65 ggc cga gtg gac ggc gac ttc ctg gag gcg gtg aag cgg cac
atc ttg 295Gly Arg Val Asp Gly Asp Phe Leu Glu Ala Val Lys Arg His
Ile Leu 70 75 80 agc cgc ctg cag atg cgg ggc cgg ccc aac atc acg
cac gcc gtg cct 343Ser Arg Leu Gln Met Arg Gly Arg Pro Asn Ile Thr
His Ala Val Pro 85 90 95 aag gcc gcc atg gtc acg gcc ctg cgc aag
ctg cac gcg ggc aag gtg 391Lys Ala Ala Met Val Thr Ala Leu Arg Lys
Leu His Ala Gly Lys Val 100 105 110 115 cgc gag gac ggc cgc gtg gag
atc ccg cac ctc gac ggc cac gcc agc 439Arg Glu Asp Gly Arg Val Glu
Ile Pro His Leu Asp Gly His Ala Ser 120 125 130 ccg ggc gcc gac ggc
cag gag cgc gtt tcc gaa atc atc agc ttc gcc 487Pro Gly Ala Asp Gly
Gln Glu Arg Val Ser Glu Ile Ile Ser Phe Ala 135 140 145 gag aca gat
ggc ctc gcc tcc tcc cgg gtc cgc cta tac ttc ttc atc 535Glu Thr Asp
Gly Leu Ala Ser Ser Arg Val Arg Leu Tyr Phe Phe Ile 150 155 160 tcc
aac gaa ggc aac cag aac ctg ttt gtg gtc cag gcc agc ctg tgg 583Ser
Asn Glu Gly Asn Gln Asn Leu Phe Val Val Gln Ala Ser Leu Trp 165 170
175 ctt tac ctg aaa ctc ctg ccc tac gtc ctg gag aag ggc agc cgg cgg
631Leu Tyr Leu Lys Leu Leu Pro Tyr Val Leu Glu Lys Gly Ser Arg Arg
180 185 190 195 aag gtg cgg gtc aaa gtg tac ttc cag gag cag ggc cac
ggt gac agg 679Lys Val Arg Val Lys Val Tyr Phe Gln Glu Gln Gly His
Gly Asp Arg 200 205 210 tgg aac atg gtg gag aag agg gtg gac ctc aag
cgc agc ggc tgg cat 727Trp Asn Met Val Glu Lys Arg Val Asp Leu Lys
Arg Ser Gly Trp His 215 220 225 acc ttc cca ctc acg gag gcc atc cag
gcc ttg ttt gag cgg ggc gag 775Thr Phe Pro Leu Thr Glu Ala Ile Gln
Ala Leu Phe Glu Arg Gly Glu 230 235 240 cgg cga ctc aac cta gac gtg
cag tgt gac agc tgc cag gag ctg gcc 823Arg Arg Leu Asn Leu Asp Val
Gln Cys Asp Ser Cys Gln Glu Leu Ala 245 250 255
gtg gtg ccg gtg ttc gtg gac cca ggc gaa gag tcg cac cgg ccc ttt
871Val Val Pro Val Phe Val Asp Pro Gly Glu Glu Ser His Arg Pro Phe
260 265 270 275 gtg gtg gtg cag gct cgg ctg ggc gac agc agg cac cgc
att cgc aag 919Val Val Val Gln Ala Arg Leu Gly Asp Ser Arg His Arg
Ile Arg Lys 280 285 290 cga ggc ctg gag tgc gat ggc cgg acc aac ctc
tgt tgc agg caa cag 967Arg Gly Leu Glu Cys Asp Gly Arg Thr Asn Leu
Cys Cys Arg Gln Gln 295 300 305 ttc ttc att gac ttc cgc ctc atc ggc
tgg aac gac tgg atc ata gca 1015Phe Phe Ile Asp Phe Arg Leu Ile Gly
Trp Asn Asp Trp Ile Ile Ala 310 315 320 ccc acc ggc tac tac ggg aac
tac tgt gag ggc agc tgc cca gcc tac 1063Pro Thr Gly Tyr Tyr Gly Asn
Tyr Cys Glu Gly Ser Cys Pro Ala Tyr 325 330 335 ctg gca ggg gtc ccc
ggc tct gcc tcc tcc ttc cac acg gct gtg gtg 1111Leu Ala Gly Val Pro
Gly Ser Ala Ser Ser Phe His Thr Ala Val Val 340 345 350 355 aac cag
tac cgc atg cgg ggt ctg aac ccc ggc acg gtg aac tcc tgc 1159Asn Gln
Tyr Arg Met Arg Gly Leu Asn Pro Gly Thr Val Asn Ser Cys 360 365 370
tgc att ccc acc aag ctg agc acc atg tcc atg ctg tac ttc gat gat
1207Cys Ile Pro Thr Lys Leu Ser Thr Met Ser Met Leu Tyr Phe Asp Asp
375 380 385 gag tac aac atc gtc aag cgg gac gtg ccc aac atg att gtg
gag gag 1255Glu Tyr Asn Ile Val Lys Arg Asp Val Pro Asn Met Ile Val
Glu Glu 390 395 400 tgc ggc tgc gcc tga cagtgcaagg caggggcacg
gtggtggggc acggagggca 1310Cys Gly Cys Ala 405 gtcccgggtg ggcttcttcc
agcccccgcg ggaacggggg tacacggtgg gctgagtaca 1370gtcattctgt
tgggctgtgg agatagtgcc agggtgcggc ctgagatatt tttctacagc
1430ttcatagagc aaccagtcaa aaccagagcg agaaccctca actgacatga
aatactttaa 1490aatgcacacg tagccacgca cagccagacg catcctgcca
cccacacagc agcctccagg 1550ataccagcaa atggatgcgg tgacaaatgg
cagcttagct acaaatgcct gtcagtcgga 1610gagaatgggg tgagcagcca
ccattcccac cagctggccc ggccactctg aattgcgcct 1670tccgagcaca
cataaaagca caaagacaga gacgcagaga gagagagaga gccacggaga
1730ggaaaagcag atgcaggggt ggggagcgca gctcggcgga ggctgcgtgt
gccccgtggc 1790ttttaccagg cctgctctgc ctggctcgat gtctgcttct
tccccagcct gggatccttc 1850gtgcttcaag gcctggggag cctgtccttc
catgcccttg tcgagggaaa gagacccaga 1910aaggacacaa cccgtcagag
acctgggagc aggggcaatg accgtttgac tgtttgtggc 1970ttgggcctct
gacatgactt atgtgtgtgt gtgtttttgg ggtggggagg gagggagaga
2030agagggggct aaatttgatg ctttaactga tctccaacag ttgacaggtc
atccttgcca 2090gttgtataac tgaaaaagga cttttctacc aggtatgacc
ttttaagtga aaatctgaat 2150tgttctaaat ggaaagaaaa aaagttgcaa
tctgtgccct tcattgggga cattcctcta 2210ggactggttt ggggacgggt
gggaatgacc cctaggcaag gggatgagac cgcaggagga 2270aatggcgggg
aggaggcatt cttgaactgc tgaggatggg gggtgtcccc tcagcggagg
2330ccaagggagg ggagcagcct agttggtctt ggagagatgg ggaaggcttt
cagctgattt 2390gcagaagttg cccatgtggg ccccagccat cagggctggc
cgtggacgtg gcccctgccc 2450actcacctgc ccgcctgccc gcccgcccgc
atagcacttg cagacctgcc tgaacgcaca 2510tgacatagca cttgccgatc
tgcgtgtgtc cagaagtggc ccttggccga gcgccgaact 2570cgctcgccct
ctagatgtcc aagtgccacg tgaactatgc aatttaaagg gttgacccac
2630actagacgaa actggactcg tacgactctt tttatatttt ttatacttga
aatgaaatcc 2690tttgcttctt ttttaagcga atgattgctt ttaatgtttg
cactgattta gttgcatgat 2750tagtcagaaa ctgccatttg aaaaaaagtt
atttttatag cagcaaaaaa aaaaaaaaaa 2810gaatacagtt aaatgtatta
tacataattt tggaaccaaa gaggccaaca gatcagtttt 2870aattttatta
gacggtgagg ccatctgaga tgaggtggac gttctgagca gtcccttgag
2930tggcctgcca acgtttcagg gtatgaatgg attttgttta ttcggtttga
tgtgtctttt 2990ccatccttac acacccagaa ggtagagtaa aaatgactat
gatagaatgc aggtgtgtat 3050ccttaaatcc tcatctttat gtttatttaa
taaagctccc cttagattct gtttcataat 3110aatttaaaac caaacaattt
tcccatagac ttgctgttaa agtattgtac gtttgtgtac 3170agtttaagaa
aataaaagat tgagtgccac gggaaaaaaa aaaaaaaa 3218436407PRTHomo sapiens
436Met Asp Gly Leu Pro Gly Arg Ala Leu Gly Ala Ala Cys Leu Leu Leu
1 5 10 15 Leu Ala Ala Gly Trp Leu Gly Pro Glu Ala Trp Gly Ser Pro
Thr Pro 20 25 30 Pro Pro Thr Pro Ala Ala Pro Pro Pro Pro Pro Pro
Pro Gly Ser Pro 35 40 45 Gly Gly Ser Gln Asp Thr Cys Thr Ser Cys
Gly Gly Phe Arg Arg Pro 50 55 60 Glu Glu Leu Gly Arg Val Asp Gly
Asp Phe Leu Glu Ala Val Lys Arg 65 70 75 80 His Ile Leu Ser Arg Leu
Gln Met Arg Gly Arg Pro Asn Ile Thr His 85 90 95 Ala Val Pro Lys
Ala Ala Met Val Thr Ala Leu Arg Lys Leu His Ala 100 105 110 Gly Lys
Val Arg Glu Asp Gly Arg Val Glu Ile Pro His Leu Asp Gly 115 120 125
His Ala Ser Pro Gly Ala Asp Gly Gln Glu Arg Val Ser Glu Ile Ile 130
135 140 Ser Phe Ala Glu Thr Asp Gly Leu Ala Ser Ser Arg Val Arg Leu
Tyr 145 150 155 160 Phe Phe Ile Ser Asn Glu Gly Asn Gln Asn Leu Phe
Val Val Gln Ala 165 170 175 Ser Leu Trp Leu Tyr Leu Lys Leu Leu Pro
Tyr Val Leu Glu Lys Gly 180 185 190 Ser Arg Arg Lys Val Arg Val Lys
Val Tyr Phe Gln Glu Gln Gly His 195 200 205 Gly Asp Arg Trp Asn Met
Val Glu Lys Arg Val Asp Leu Lys Arg Ser 210 215 220 Gly Trp His Thr
Phe Pro Leu Thr Glu Ala Ile Gln Ala Leu Phe Glu 225 230 235 240 Arg
Gly Glu Arg Arg Leu Asn Leu Asp Val Gln Cys Asp Ser Cys Gln 245 250
255 Glu Leu Ala Val Val Pro Val Phe Val Asp Pro Gly Glu Glu Ser His
260 265 270 Arg Pro Phe Val Val Val Gln Ala Arg Leu Gly Asp Ser Arg
His Arg 275 280 285 Ile Arg Lys Arg Gly Leu Glu Cys Asp Gly Arg Thr
Asn Leu Cys Cys 290 295 300 Arg Gln Gln Phe Phe Ile Asp Phe Arg Leu
Ile Gly Trp Asn Asp Trp 305 310 315 320 Ile Ile Ala Pro Thr Gly Tyr
Tyr Gly Asn Tyr Cys Glu Gly Ser Cys 325 330 335 Pro Ala Tyr Leu Ala
Gly Val Pro Gly Ser Ala Ser Ser Phe His Thr 340 345 350 Ala Val Val
Asn Gln Tyr Arg Met Arg Gly Leu Asn Pro Gly Thr Val 355 360 365 Asn
Ser Cys Cys Ile Pro Thr Lys Leu Ser Thr Met Ser Met Leu Tyr 370 375
380 Phe Asp Asp Glu Tyr Asn Ile Val Lys Arg Asp Val Pro Asn Met Ile
385 390 395 400 Val Glu Glu Cys Gly Cys Ala 405
* * * * *
References