U.S. patent application number 14/810476 was filed with the patent office on 2015-11-12 for tolerogenic synthetic nanocarriers for inducing regulatory b cells.
This patent application is currently assigned to Selecta Biosciences, Inc.. The applicant listed for this patent is Selecta Biosciences, Inc.. Invention is credited to Roberto A. Maldonado.
Application Number | 20150320870 14/810476 |
Document ID | / |
Family ID | 47068065 |
Filed Date | 2015-11-12 |
United States Patent
Application |
20150320870 |
Kind Code |
A1 |
Maldonado; Roberto A. |
November 12, 2015 |
TOLEROGENIC SYNTHETIC NANOCARRIERS FOR INDUCING REGULATORY B
CELLS
Abstract
Disclosed are synthetic nanocarrier methods, and related
compositions, comprising B cell and/or MHC Class II-restricted
epitopes and immunosuppressants in order to generate tolerogenic
immune responses, such as the generation of antigen-specific
regulatory B cells.
Inventors: |
Maldonado; Roberto A.;
(Jamaica Plain, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Selecta Biosciences, Inc. |
Watertown |
MA |
US |
|
|
Assignee: |
Selecta Biosciences, Inc.
Watertown
MA
|
Family ID: |
47068065 |
Appl. No.: |
14/810476 |
Filed: |
July 27, 2015 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14161660 |
Jan 22, 2014 |
|
|
|
14810476 |
|
|
|
|
13458179 |
Apr 27, 2012 |
8652487 |
|
|
14161660 |
|
|
|
|
61531147 |
Sep 6, 2011 |
|
|
|
61531153 |
Sep 6, 2011 |
|
|
|
61531164 |
Sep 6, 2011 |
|
|
|
61531168 |
Sep 6, 2011 |
|
|
|
61531175 |
Sep 6, 2011 |
|
|
|
61531180 |
Sep 6, 2011 |
|
|
|
61531194 |
Sep 6, 2011 |
|
|
|
61531204 |
Sep 6, 2011 |
|
|
|
61531209 |
Sep 6, 2011 |
|
|
|
61531215 |
Sep 6, 2011 |
|
|
|
61513514 |
Jul 29, 2011 |
|
|
|
61480946 |
Apr 29, 2011 |
|
|
|
Current U.S.
Class: |
424/184.1 ;
435/325 |
Current CPC
Class: |
A61P 37/00 20180101;
A61K 9/5115 20130101; A61P 37/02 20180101; A61K 9/5146 20130101;
A61K 47/52 20170801; B82Y 40/00 20130101; A61K 9/14 20130101; A61K
2039/577 20130101; A61K 9/51 20130101; A61K 2039/55555 20130101;
G01N 33/56972 20130101; A61P 7/06 20180101; A61K 39/35 20130101;
A61K 47/50 20170801; A61P 37/06 20180101; G01N 33/505 20130101;
A61K 39/385 20130101; G01N 2333/70517 20130101; A61K 47/6929
20170801; B82Y 5/00 20130101; A61P 11/06 20180101; A61K 39/0008
20130101; A61K 39/36 20130101; A61K 47/6923 20170801; A61P 1/16
20180101; A61K 38/13 20130101; A61P 29/00 20180101; A61K 39/001
20130101; A61K 47/544 20170801; G01N 2333/7051 20130101; A61K
31/192 20130101; A61K 47/6937 20170801; A61P 11/02 20180101; A61P
35/00 20180101; A61K 38/38 20130101; A61K 38/1816 20130101; A61K
47/643 20170801; A61K 9/127 20130101; A61K 47/69 20170801; A61P
43/00 20180101; Y02A 50/30 20180101; A61P 37/04 20180101; A61P
37/08 20180101; A61K 9/5153 20130101; A61K 39/00 20130101; G01N
2333/70514 20130101; A61K 2039/55511 20130101; A61K 47/593
20170801; A61P 41/00 20180101; A61K 2039/5154 20130101; A61P 15/00
20180101; A61P 17/00 20180101; A61K 39/36 20130101; A61K 2300/00
20130101 |
International
Class: |
A61K 47/48 20060101
A61K047/48; A61K 39/00 20060101 A61K039/00 |
Claims
1. A method comprising: (A) administering to a subject a
composition that comprises: (i) a first population of synthetic
nanocarriers coupled to immunosuppressants, and (ii) B cell and/or
MHC Class II-restricted epitopes of an antigen, wherein the
composition is in an amount effective to generate antigen-specific
regulatory B cells in the subject; or (B) generating
antigen-specific regulatory B cells in a subject by administering a
composition that comprises: (i) a first population of synthetic
nanocarriers coupled to immunosuppressants, and (ii) B cell and/or
MHC Class II-restricted epitopes of an antigen; or (C)
administering to a subject a composition according to a protocol
that was previously shown to generate antigen-specific regulatory B
cells in one or more test subjects; wherein the composition
comprises: (i) a first population of synthetic nanocarriers coupled
to immunosuppressants, and (ii) B cell and/or MHC Class
II-restricted epitopes of an antigen.
2-39. (canceled)
40. A composition comprising isolated antigen-specific regulatory B
cells produced according to the method of claim 1.
41-42. (canceled)
43. A dosage form comprising the composition of claim 40.
44-45. (canceled)
46. A composition comprising: (i) a first population of synthetic
nanocarriers coupled to immunosuppressants, and (ii) B cell and/or
MHC Class II-restricted epitopes of an antigen, for use in therapy
or prophylaxis.
47-49. (canceled)
50. Use of the composition of claim 40 for the manufacture of a
medicament for use in promoting a tolerogenic immune response or
generating antigen-specific regulatory B cells in a subject.
51. (canceled)
52. A method comprising: (i) producing a first population of
synthetic nanocarriers coupled to immunosuppressants, and (ii)
producing or obtaining B cell and/or MHC Class II-restricted
epitopes of an antigen, wherein the composition is in an amount
effective to generate antigen-specific regulatory B cells in a
subject.
53-60. (canceled)
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 14/161,660, filed Jan. 22, 2014, pending,
which is a continuation of U.S. patent application Ser. No.
13/458,179, filed Apr. 27, 2012, granted as U.S. Pat. No. 8,652,487
on Feb. 18, 2014, which claims the benefit under 35 U.S.C.
.sctn.119 of U.S. Provisional Patent Application 61/480,946, filed
Apr. 29, 2011, 61/513,514, filed Jul. 29, 2011, 61/531,147, filed
Sep. 6, 2011, 61/531,153, filed Sep. 6, 2011, 61/531,164, filed
Sep. 6, 2011, 61/531,168, filed Sep. 6, 2011, 61/531,175, filed
Sep. 6, 2011, 61/531,180, filed Sep. 6, 2011, 61/531,194, filed
Sep. 6, 2011, 61/531,204, filed Sep. 6, 2011, 61/531,209, filed
Sep. 6, 2011, 61/531,215, filed Sep. 6, 2011, the entire contents
of each of which are incorporated herein by reference.
FIELD OF THE INVENTION
[0002] This invention relates to methods of administering synthetic
nanocarrier compositions with immunosuppressants and B cell and/or
MHC Class II-restricted epitopes of an antigen wherein the
compositions can generate regulatory B cells, and related
compositions. The methods allow for efficient uptake by APCs to
shift the immune response in favor of regulatory B cell development
specific to antigens.
BACKGROUND OF THE INVENTION
[0003] Conventional strategies for generating immunosuppression
associated with an undesired immune response are based on
broad-acting immunosuppressive drugs. Additionally, in order to
maintain immunosuppression, immunosuppressant drug therapy is
generally a life-long proposition. Unfortunately, the use of
broad-acting immunosuppressants are associated with a risk of
severe side effects, such as tumors, infections, nephrotoxicity and
metabolic disorders. Accordingly, new immunosuppressant therapies
would be beneficial.
SUMMARY OF THE INVENTION
[0004] In one aspect, a method comprising administering to a
subject a composition that comprises: (i) a first population of
synthetic nanocarriers coupled to immunosuppressants, and (ii) a
second population of synthetic nanocarriers coupled to comprise B
cell and/or MHC Class II-restricted epitopes of an antigen is
provided. In another aspect, a method comprising generating
antigen-specific regulatory B cells in a subject by administering a
composition that comprises (i) a first population of synthetic
nanocarriers coupled to immunosuppressants, and (ii) a second
population of synthetic nanocarriers coupled to B cell and/or MHC
Class II-restricted epitopes of an antigen is provided. In another
aspect, a method comprising: administering to a subject a
composition according to a protocol that was previously shown to
generate antigen-specific regulatory B cells in one or more test
subjects; wherein the composition comprises: (i) a first population
of synthetic nanocarriers coupled to immunosuppressants, and (ii) a
second population of synthetic nanocarriers coupled to B cell
and/or MHC Class II-restricted epitopes of an antigen is provided.
In one embodiment, the composition is in an amount effective to
generate antigen-specific regulatory B cells in the subject. In
another embodiment, the composition is administered in an amount
effective to generate antigen-specific regulatory B cells in the
subject.
[0005] In one embodiment, the first population and the second
population are the same population. In another embodiment, the
first population and second population are different
populations.
[0006] In another embodiment, the method further comprises
providing or identifying the subject.
[0007] In another embodiment, the antigen is a therapeutic protein,
an autoantigen or an allergen, or is associated with an
inflammatory disease, an autoimmune disease, organ or tissue
rejection or graft versus host disease.
[0008] In another embodiment, the method further comprises
assessing the generation of antigen-specific regulatory B cells in
the subject prior to and/or after the administration of the
composition.
[0009] In another embodiment, the subject has or is at risk of
having an inflammatory disease, an autoimmune disease, an allergy,
organ or tissue rejection or graft versus host disease. In another
embodiment, the subject has undergone or will undergo
transplantation. In another embodiment, the subject has or is at
risk of having an undesired immune response against a therapeutic
protein that is being administered or will be administered to the
subject.
[0010] In another embodiment, one or more maintenance doses of the
composition comprising the first population and second population
of synthetic nanocarriers is administered to the subject.
[0011] In another embodiment, the administering is by intravenous,
intraperitoneal, transmucosal, oral, subcutaneous, pulmonary,
intranasal, intradermal or intramuscular administration. In another
embodiment, the administering is by inhalation or intravenous,
subcutaneous or transmucosal administration.
[0012] In another embodiment, the method further comprises
administering a transplantable graft or therapeutic protein. In
another embodiment, the administering of the transplantable graft
or therapeutic protein, when the therapeutic protein is provided as
one or more cells, is by parenteral, intraarterial, intranasal or
intravenous administration or by injection to lymph nodes or
anterior chamber of the eye or by local administration to an organ
or tissue of interest.
[0013] In another embodiment, the method further comprises
collecting the generated antigen-specific regulatory B cells.
[0014] In another embodiment, the immunosuppressants comprise a
statin, an mTOR inhibitor, a TGF-.beta. signaling agent, a
corticosteroid, an inhibitor of mitochondrial function, a P38
inhibitor, an NF-.kappa..beta. inhibitor, an adenosine receptor
agonist, a prostaglandin E2 agonist, a phosphodiesterasse 4
inhibitor, an HDAC inhibitor or a proteasome inhibitor. In another
embodiment, the mTOR inhibitor is rapamycin or a rapamycin
analog.
[0015] In another embodiment, the load of the immunosuppressants
and/or antigens on average across the first and/or second
population of synthetic nanocarriers is between 0.0001% and 50%
(weight/weight). In another embodiment, the load of the
immunosuppressants and/or antigens on average across the first
and/or second population of synthetic nanocarriers is between 0.1%
and 10% (weight/weight).
[0016] In another embodiment, the synthetic nanocarriers of the
first population and/or second population comprise lipid
nanoparticles, polymeric nanoparticles, metallic nanoparticles,
surfactant-based emulsions, dendrimers, buckyballs, nanowires,
virus-like particles or peptide or protein particles. In another
embodiment, the synthetic nanocarriers of the first population
and/or second population comprise lipid nanoparticles. In another
embodiment, the synthetic nanocarriers of the first population
and/or second population comprise liposomes. In another embodiment,
the synthetic nanocarriers of the first population and/or second
population comprise metallic nanoparticles. In another embodiment,
the metallic nanoparticles comprise gold nanoparticles. In another
embodiment, the synthetic nanocarriers of the first population
and/or second population comprise polymeric nanoparticles. In
another embodiment, the polymeric nanoparticle comprises polymer
that is a non-methoxy-terminated, pluronic polymer. In another
embodiment, the polymeric nanoparticles comprise a polyester, a
polyester coupled to a polyether, polyamino acid, polycarbonate,
polyacetal, polyketal, polysaccharide, polyethyloxazoline or
polyethyleneimine. In another embodiment, the polyester comprises a
poly(lactic acid), poly(glycolic acid), poly(lactic-co-glycolic
acid) or polycaprolactone. In another embodiment, the polymeric
nanoparticles comprise a polyester and a polyester coupled to a
polyether. In another embodiment, the polyether comprises
polyethylene glycol or polypropylene glycol.
[0017] In another embodiment, the mean of a particle size
distribution obtained using dynamic light scattering of the
synthetic nanocarriers of the first and/or second population is a
diameter greater than 100 nm. In another embodiment, the diameter
is greater than 150 nm. In another embodiment, the diameter is
greater than 200 nm. In another embodiment, the diameter is greater
than 250 nm. In another embodiment, the diameter is greater than
300 nm. In another embodiment, the aspect ratio of the synthetic
nanocarriers of the first population and/or second population is
greater than 1:1, 1:1.2, 1:1.5, 1:2, 1:3, 1:5, 1:7 or 1:10.
[0018] In another embodiment, the method further comprises making a
dosage form comprising the collected antigen-specific regulatory B
cells. In another embodiment, the method further comprises making
the collected antigen-specific regulatory B cells available or
dosage form available to a subject for administration.
[0019] In another embodiment, the second population of synthetic
nanocarriers are also coupled to antigens that comprise MHC
I-restricted epitopes.
[0020] In another aspect, a composition comprising the isolated
antigen-specific regulatory B cells produced according to any of
the methods provided herein is provided.
[0021] In another embodiment, the composition further comprises a
pharmaceutically acceptable excipient.
[0022] In another aspect, a dosage form comprising the any of the
compositions provided herein is provided.
[0023] In another aspect, a composition comprising isolated
antigen-specific regulatory B cells for use in therapy or
prophylaxis, said isolated antigen-specific regulatory B cells
being obtainable by a process comprising the steps of: (a)
generating antigen-specific regulatory B cells in a subject by
administering a composition that comprises: (i) a first population
of synthetic nanocarriers coupled to immunosuppressants, and (ii) a
second population of synthetic nanocarriers coupled to B cell
and/or MHC Class II-restricted epitopes of an antigen; and (b)
collecting the generated antigen-specific regulatory B cells is
provided. In one embodiment, the composition is any of the
compositions provided herein or used in any of the methods
provided.
[0024] In another aspect, a composition comprising: (i) a first
population of synthetic nanocarriers coupled to immunosuppressants,
and (ii) a second population of synthetic nanocarriers coupled to B
cell and/or MHC Class II-restricted epitopes of an antigen, for use
in therapy or prophylaxis is provided. In one embodiment, the
composition is any of the compositions provided herein or used in
any of the methods provided.
[0025] In another aspect, any of the compositions provided may be
for use in therapy or prophylaxis.
[0026] In another aspect, any of the compositions provided may be
for use in promoting a tolerogenic immune response or generating
antigen-specific regulatory B cells in a subject.
[0027] In another aspect, a use of any of the compositions or
dosage forms provided for the manufacture of a medicament for use
in promoting a tolerogenic immune response or generating
antigen-specific regulatory B cells in a subject is provided.
[0028] In another embodiment, the composition further comprises a
transplantable graft or therapeutic protein.
[0029] In another aspect, a dosage form comprising any of the
compositions provided is provided.
[0030] In another aspect, a method comprising: (i) producing a
first population of synthetic nanocarriers coupled to
immunosuppressants, and (ii) producing a second population of
synthetic nanocarriers coupled to B cell and/or MHC Class
II-restricted epitopes of an antigen is provided.
[0031] In one embodiment, the composition is in an amount effective
to generate antigen-specific regulatory B cells in a subject. In
another embodiment, the first population and second population are
the same population. In another embodiment, the first population
and second population are different populations.
[0032] In another embodiment, the method further comprises making a
dosage form comprising the first population and second population
of synthetic nanocarriers.
[0033] In another embodiment, the method further comprises making a
composition comprising the first population and second population
of synthetic nanocarriers or the dosage form available to a subject
for administration.
[0034] In another embodiment, the method further comprises
assessing the generation of antigen-specific regulatory B cells
with a composition comprising the first population and second
population of synthetic nanocarriers.
[0035] In another embodiment, the first population and second
population of synthetic nanocarriers that are produced are as
defined in any of the compositions and methods provided herein.
[0036] In another aspect, a process for producing a composition or
dosage form comprising the steps of: (i) coupling a first
population of synthetic nanocarriers to immunosuppressants, and
(ii) coupling a second population of synthetic nanocarriers to B
cell and/or MHC Class II-restricted epitopes of an antigen is
provided. In one embodiment, the composition or dosage form is in
an amount effective to generate regulatory B cells in a subject. In
one embodiment, the steps are as defined in any of the methods
provided herein.
[0037] In another aspect, a composition or dosage form obtainable
by any of the methods or processes provided herein is provided.
[0038] In another embodiment, the methods further comprise
including a transplantable graft or therapeutic protein with the
collected antigen-specific regulatory B cells or dosage form.
[0039] In another embodiment, the first and/or second population of
synthetic nanocarriers of any of the compositions or methods
provided are also coupled to MHC Class I-restricted epitopes. In an
embodiment of any of the compositions and methods provided herein,
the antigens comprise substantially no MHC Class I-restricted
epitopes.
[0040] In an embodiment of any of the compositions and methods
provided herein, antigens that are proteins that comprise the
aforementioned epitopes can be coupled to the synthetic
nanocarriers. In another embodiment, polypeptides or peptides that
comprise the aforementioned epitopes but additional amino acids
that flank one or both ends of the epitope(s) can be coupled to the
synthetic nanocarriers. In another embodiment, the epitopes
themselves are coupled to the synthetic nanocarriers.
BRIEF DESCRIPTION OF FIGURES
[0041] FIG. 1 provides a representative example of a flow
cytometric analysis of Treg cells.
[0042] FIG. 2 demonstrates that synthetic nanocarriers of the
invention resulted in higher proportions of antigen-specific
Bregs.
[0043] FIG. 3 shows a decrease in IgE production with synthetic
nanocarriers of the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0044] Before describing the present invention in detail, it is to
be understood that this invention is not limited to particularly
exemplified materials or process parameters as such may, of course,
vary. It is also to be understood that the terminology used herein
is for the purpose of describing particular embodiments of the
invention only, and is not intended to be limiting of the use of
alternative terminology to describe the present invention.
[0045] All publications, patents and patent applications cited
herein, whether supra or infra, are hereby incorporated by
reference in their entirety for all purposes.
[0046] As used in this specification and the appended claims, the
singular forms "a," "an" and "the" include plural referents unless
the content clearly dictates otherwise. For example, reference to
"a polymer" includes a mixture of two or more such molecules or a
mixture of differing molecular weights of a single polymer species,
reference to "a synthetic nanocarrier" includes a mixture of two or
more such synthetic nanocarriers or a plurality of such synthetic
nanocarriers, reference to "a DNA molecule" includes a mixture of
two or more such DNA molecules or a plurality of such DNA
molecules, reference to "an immunosuppressant" includes a mixture
of two or more such materials or a plurality of an
immunosuppressant molecules, and the like.
[0047] As used herein, the term "comprise" or variations thereof
such as "comprises" or "comprising" are to be read to indicate the
inclusion of any recited integer (e.g. a feature, element,
characteristic, property, method/process step or limitation) or
group of integers (e.g. features, element, characteristics,
properties, method/process steps or limitations) but not the
exclusion of any other integer or group of integers. Thus, as used
herein, the term "comprising" is inclusive and does not exclude
additional, unrecited integers or method/process steps.
[0048] In embodiments of any of the compositions and methods
provided herein, "comprising" may be replaced with "consisting
essentially of" or "consisting of". The phrase "consisting
essentially of" is used herein to require the specified integer(s)
or steps as well as those which do not materially affect the
character or function of the claimed invention. As used herein, the
term "consisting" is used to indicate the presence of the recited
integer (e.g. a feature, element, characteristic, property,
method/process step or limitation) or group of integers (e.g.
features, element, characteristics, properties, method/process
steps or limitations) alone.
A. INTRODUCTION
[0049] Delivering immunosuppressants and antigens via synthetic
nanocarrier compositions that comprise B cell and/or MHC Class
II-restricted epitopes more directly at the sites of action in
cells of interest, in particular APCs, can stimulate regulatory B
cells and result in beneficial tolerogenic immune responses
specific to the antigens. As shown in the examples, synthetic
nanocarrier compositions were found to be effective in generating
CD24+ B cells that produce IL-10 and TGF-.beta. and also recognize
antigen, which speaks to the antigen-specific nature of the
response. Additionally, the same synthetic nanocarrier compositions
were found to decrease IgE production, which suggests a downstream
result of the regulatory B cell production. The compositions
provided, without being bound to any particular theory, can provide
tolerogenic immune effects directly as a result of the recognition
of epitopes, which may cause the generation of the regulatory
cells, the switching of B cells to a regulatory phenotype, etc. The
tolerogenic immune responses may also be a result of the production
of cytokines and/or the production of other regulatory immune cells
stimulated by such cytokines. The generation of regulatory B cells
can also lead to downstream tolerogenic effects. For example,
regulatory B cells can reduce the proliferative capacity of
effector T cells, such as CD4+ T cells, and/or enhance FoxP3 and
CTLA-4 expression in regulatory T cells. Regulatory B cells can
also produce the regulatory cytokine, IL-10. In embodiments, the
antigen-specific itDCs provided herein may have these effects, for
example, on a subject.
[0050] The generation of such regulatory B cells evidences the
ability of the compositions of the invention to generate
antigen-specific tolerogenic immune responses that can have utility
in the treatment or prophylaxis of a variety of diseases, disorders
or conditions. Therefore, this invention is useful, for example, to
promote tolerogenic immune responses in subjects who have or are at
risk of having an allergy, autoimmune disease, an inflammatory
disease, organ or tissue rejection or graft versus host disease.
This invention is also useful for promoting tolerogenic immune
responses in subjects who have undergone or will undergo
transplantation. This invention is also useful for promoting
tolerogenic immune responses in subjects that have received, are
receiving or will receive a therapeutic protein against which
undesired immune responses are generated or are expected to be
generated. The present invention, in some embodiments, prevents or
suppresses undesired immune responses that may neutralize the
beneficial effect of certain therapeutic treatments.
[0051] The inventors have unexpectedly and surprisingly discovered
that the problems and limitations noted above can be overcome by
practicing the invention disclosed herein. In particular, the
inventors have unexpectedly discovered that it is possible to
provide synthetic nanocarrier compositions, and related methods,
that induce a tolerogenic immune response through the the increase
in the number and/or activity of regulatory B cells. The method
described herein include administering to a subject a composition
that comprises (i) a first population of synthetic nanocarriers
coupled to immunosuppressants, and (ii) a second population of
synthetic nanocarriers coupled to B cell and/or MHC Class
II-restricted epitopes of an antigen. Preferably, the composition
is in an amount effective to effect antigen-specific regulatory B
cell proliferation and/or activity in the subject. In another
aspect, a method comprising generating antigen-specific regulatory
B cells in a subject by administering a composition that comprises
(i) a first population of synthetic nanocarriers coupled to
immunosuppressants, and (ii) a second population of synthetic
nanocarriers coupled to B cell and/or MHC Class II-restricted
epitopes of an antigen is provided. In another aspect, a method
comprising administering to a subject a composition according to a
protocol that was previously shown to generate antigen-specific
regulatory B cells in one or more test subjects wherein the
composition comprises (i) a first population of synthetic
nanocarriers coupled to immunosuppressants, and (ii) a second
population of synthetic nanocarriers coupled to B cell and/or MHC
Class II-restricted epitopes of an antigen is provided.
[0052] Transplantable grafts, therapeutic proteins, etc. may also
be administered to the subjects as provided herein. Such
compositions may be administered to a subject prior to,
concomitantly with or after the administration of the first and
second populations of synthetic nanocarriers. Such additional
agents may or may not be coupled to the first or second population
of synthetic nanocarriers or another population of synthetic
nanocarriers. In embodiments, the compositions provided may also be
administered as one or more maintenance doses to a subject. In such
embodiments, the compositions provided are administered such that
the generation of a desired immune response is maintained or an
undesired immune response is reduced for a certain length of time.
Examples of such lengths of time are provided elsewhere herein.
[0053] In another aspect, a composition comprising isolated
antigen-specific regulatory B cells produced according to any of
the methods provided is provided. In yet another aspect, the
compositions comprising the first population and second population
of synthetic nanocarriers are also provided. In embodiments, the
compositions further comprise a transplantable graft or a
therapeutic protein that may or may not be coupled to the first or
second population of synthetic nanocarriers or another population
of synthetic nanocarriers.
[0054] In another aspect, dosage forms of any of the compositions
herein are provided. Such dosage forms may be administered to a
subject, for example, one in need of antigen-specific regulatory B
cell generation.
[0055] In yet another aspect, a method of (i) producing a first
population of synthetic nanocarriers coupled to immunosuppressants,
and (ii) producing a second population of synthetic nanocarriers
coupled to B cell and/or MHC Class II-restricted epitopes of an
antigen is provided. In one embodiment, the method further
comprises producing a dosage form comprising the first and second
populations of synthetic nanocarriers. In another embodiment, the
method further comprises ensuring the second population of
synthetic nanocarriers comprises B cell and/or MHC Class
II-restricted epitopes of an antigen. In one embodiment, the
presence of a B cell epitope and/or MHC Class II-restricted can be
confirmed by the ability of such epitope to stimulate immune
responses such as antibody production or B cell proliferation
and/or activity for the B cell epitopes and CD4+ T cell immune
responses for the MHC Class II-restricted epitopes. Methods for
determining these activity are well known to those of ordinary
skill in the art or are provided elsewhere herein. In still another
embodiment, the method further comprises assessing the generation
of regulatory B cells with a composition or dosage form comprising
the first population and second population of synthetic
nanocarriers. In one embodiment, this assessment is performed in
vitro. In another embodiment, this assessment is performed in vivo,
such as with a sample obtained from a subject. In yet another
embodiment, the method further comprises making a composition
comprising the first population and second population of synthetic
nanocarriers or the dosage form available to a subject for
administration.
[0056] In another aspect, the compositions or dosage forms produced
by any of the methods provided herein are also provided.
[0057] The invention will now be described in more detail
below.
B. DEFINITIONS
[0058] "Administering" or "administration" means providing a
material to a subject in a manner that is pharmacologically
useful.
[0059] "Allergens" are any substances that can cause an undesired
(e.g., a Type 1 hypersensitive) immune response (i.e., an allergic
response or reaction) in a subject. Allergens include, but are not
limited to, plant allergens (e.g., pollen, ragweed allergen),
insect allergens, insect sting allergens (e.g., bee sting
allergens), animal allergens (e.g., pet allergens, such as animal
dander or cat Fel d 1 antigen), latex allergens, mold allergens,
fungal allergens, cosmetic allergens, drug allergens, food
allergens, dust, insect venom, viruses, bacteria, etc. Food
allergens include, but are not limited to milk allergens, egg
allergens, nut allergens (e.g., peanut or tree nut allergens, etc.
(e.g., walnuts, cashews, etc.)), fish allergens, shellfish
allergens, soy allergens, legume allergens, seed allergens and
wheat allergens. Insect sting allergens include allergens that are
or are associated with bee stings, wasp stings, hornet stings,
yellow jacket stings, etc. Insect allergens also include house dust
mite allergens (e.g., Der P1 antigen) and cockroach allergens. Drug
allergens include allergens that are or are associated with
antibiotics, NSAIDs, anaesthetics, etc. Pollen allergens include
grass allergens, tree allergens, weed allergens, flower allergens,
etc. Subjects that develop or are at risk of developing an
undesired immune response to any of the allergens provided herein
may be treated with any of the compositions and methods provided
herein. Subjects that may be treated with any of the compositions
and methods provided also include those who have or are at risk of
having an allergy to any of the allergens provided.
[0060] An "allergy" also referred to herein as an "allergic
condition," is any condition where there is an undesired (e.g., a
Type 1 hypersensitive) immune response (i.e., allergic response or
reaction) to a substance. Such substances are referred to herein as
allergens. Allergies or allergic conditions include, but are not
limited to, allergic asthma, hay fever, hives, eczema, plant
allergies, bee sting allergies, pet allergies, latex allergies,
mold allergies, cosmetic allergies, food allergies, allergic
rhinitis or coryza, topic allergic reactions, anaphylaxis, atopic
dermatitis, hypersensitivity reactions and other allergic
conditions. The allergic reaction may be the result of an immune
reaction to any allergen. In some embodiments, the allergy is a
food allergy. Food allergies include, but are not limited to, milk
allergies, egg allergies, nut allergies, fish allergies, shellfish
allergies, soy allergies or wheat allergies.
[0061] "Amount effective" in the context of a composition or dosage
form for administration to a subject refers to an amount of the
composition or dosage form that produces one or more desired immune
responses in the subject, for example, the generation of regulatory
B cells. Therefore, in some embodiments, an amount effective is any
amount of a composition provided herein that produces such a
desired immune response. This amount can be for in vitro or in vivo
purposes. For in vivo purposes, the amount can be one that a
clinician would believe may have a clinical benefit for a subject
in need of antigen-specific tolerization. Such subjects include
those that have or are at risk of having an inflammatory disease,
an autoimmune disease, an allergy, organ or tissue rejection or
graft versus host disease. Such subjects also include those that
have undergone or will undergo transplantation. Such subjects
further include those that have experienced, are experiencing or
are expected to experience an undesired immune response against a
therapeutic protein. Other subjects include those described
elsewhere herein.
[0062] Amounts effective can involve only reducing the level of an
undesired immune response, although in some embodiments, it
involves preventing an undesired immune response altogether.
Amounts effective can also involve delaying the occurrence of an
undesired immune response. Amounts effective can also involve
generating a desired immune response. An amount that is effective
can also be an amount of a composition provided herein that
produces a desired therapeutic endpoint or a desired therapeutic
result. Amounts effective, preferably, result in a tolerogenic
immune response in a subject to an antigen. Preferably, the amounts
effective result in the increased proliferation and/or activity of
regulatory B cells. The achievement of any of the foregoing can be
monitored by routine methods.
[0063] In some embodiments of any of the compositions and methods
provided, the amount effective is one in which the desired immune
response persists in the subject for at least 1 week, at least 2
weeks, at least 1 month, at least 2 months, at least 3 months, at
least 4 months, at least 5 months, at least 6 months, at least 9
months, at least 1 year, at least 2 years, at least 5 years, or
longer. In other embodiments of any of the compositions and methods
provided, the amount effective is one which produces a measurable
desired immune response, for example, a measurable decrease in an
immune response (e.g., to a specific antigen), for at least 1 week,
at least 2 weeks, at least 1 month, at least 2 months, at least 3
months, at least 4 months, at least 5 months, at least 6 months, at
least 9 months, at least 1 year, at least 2 years, at least 5
years, or longer.
[0064] Amounts effective will depend, of course, on the particular
subject being treated; the severity of a condition, disease or
disorder; the individual patient parameters including age, physical
condition, size and weight; the duration of the treatment; the
nature of concurrent therapy (if any); the specific route of
administration and like factors within the knowledge and expertise
of the health practitioner. These factors are well known to those
of ordinary skill in the art and can be addressed with no more than
routine experimentation. It is generally preferred that a maximum
dose be used, that is, the highest safe dose according to sound
medical judgment. It will be understood by those of ordinary skill
in the art, however, that a patient may insist upon a lower dose or
tolerable dose for medical reasons, psychological reasons or for
virtually any other reason.
[0065] In general, doses of the immunosuppressants and/or antigens
in the compositions of the invention can range from about 10
.mu.g/kg to about 100,000 .mu.g/kg. In some embodiments, the doses
can range from about 0.1 mg/kg to about 100 mg/kg. In still other
embodiments, the doses can range from about 0.1 mg/kg to about 25
mg/kg, about 25 mg/kg to about 50 mg/kg, about 50 mg/kg to about 75
mg/kg or about 75 mg/kg to about 100 mg/kg. Alternatively, the dose
can be administered based on the number of synthetic nanocarriers
that provide the desired amount of immunosuppressants and/or
antigens. For example, useful doses include greater than 10.sup.6,
10.sup.7, 10.sup.8, 10.sup.9 or 10.sup.10 synthetic nanocarriers
per dose. Other examples of useful doses include from about
1.times.10.sup.6 to about 1.times.10.sup.10, about 1.times.10.sup.7
to about 1.times.10.sup.9 or about 1.times.10.sup.8 to about
1.times.10.sup.9 synthetic nanocarriers per dose.
[0066] "Antigen" means a B cell antigen or T cell antigen. "Type(s)
of antigens" means molecules that share the same, or substantially
the same, antigenic characteristics. In some embodiments, antigens
may be proteins, polypeptides, peptides, lipoproteins, glycolipids,
polynucleotides, polysaccharides or are contained or expressed in
cells. In some embodiments, such as when the antigens are not well
defined or characterized, the antigens may be contained within a
cell or tissue preparation, cell debris, cell exosomes, conditioned
media, etc. An antigen can be combined with the synthetic
nanocarriers in the same form as what a subject is exposed to that
causes an undesired immune response but may also be a fragment or
derivative thereof. When a fragment or derivative, however, a
desired immune response to the form encountered by such a subject
is the preferable result with the compositions and methods
provided. Preferably, the antigens comprise epitopes such that
regulatory B cells are generated, recruited or activated with the
compositions provided herein. Whether or not this occurs can be
established through measurement of cytokine production (e.g.,
IL-10) and/or regulatory B cell proliferation using conventional
techniques.
[0067] "Antigen-specific" refers to any immune response that
results from the presence of the antigen, or portion thereof, or
that generates molecules that specifically recognize or bind the
antigen. For example, where the immune response is antigen-specific
antibody production, antibodies are produced that specifically bind
the antigen. As another example, where the immune response is
antigen-specific regulatory B cell proliferation and/or activity,
the proliferation and/or activity results from recognition of the
antigen, or portion thereof, alone or in complex with MHC
molecules, by B cells, etc.
[0068] "Antigens associated" with a disease, disorder or condition
provided herein are antigens that can generate an undesired immune
response against, as a result of, or in conjunction with the
disease, disorder or condition; the cause of the disease, disorder
or condition (or a symptom or effect thereof); and/or can generate
an undesired immune response that is a symptom, result or effect of
the disease, disorder or condition. Preferably, in some
embodiments, the use of an antigen associated with a disease,
disorder or condition, etc. in the compositions and methods
provided herein will lead to a tolerogenic immune response against
the antigen and/or the cells, by, on or in which the antigen is
expressed. The antigens can be in the same form as expressed in a
subject with the disease, disorder or condition but may also be a
fragment or derivative thereof. When a fragment or derivative,
however, a desired immune response to the form expressed in such a
subject is the preferable result with the compositions and methods
provided.
[0069] In one embodiment, the antigen is an antigen associated with
an inflammatory disease, autoimmune disease, organ or tissue
rejection or graft versus host disease. Such antigens include
autoantigens, such as myelin basic protein, collagen (e.g.,
collagen type 11), human cartilage gp 39, chromogranin A,
gp130-RAPS, proteolipid protein, fibrillarin, nuclear proteins,
nucleolar proteins (e.g., small nucleolar protein), thyroid
stimulating factor receptor, histones, glycoprotein gp 70,
ribosomal proteins, pyruvate dehydrogenase dehydrolipoamide
acetyltransferase, hair follicle antigens, human tropomyosin
isoform 5, mitochondrial proteins, pancreatic .beta.-cell proteins,
myelin oligodendrocyte glycoprotein, insulin, glutamic acid
decarboxylase (GAD), gluten, and fragments or derivatives thereof.
Other autoantigens are provided in Table 1 below.
[0070] Antigens also include those associated with organ or tissue
rejection. Examples of such antigens include, but are not limited
to, antigens from allogeneic cells, e.g., antigens from an
allogeneic cell extract and antigens from other cells, such as
endothelial cell antigens.
[0071] Antigens also include those associated with an allergy. Such
antigens include the allergens described elsewhere herein.
[0072] Antigens also include those associated with a transplantable
graft. Such antigens are associated with a transplantable graft, or
an undesired immune response in a recipient of a transplantable
graft that is generated as a result of the introduction of the
transplantable graft in the recipient, that can be presented for
recognition by cells of the immune system and that can generate an
undesired immune response. Transplant antigens include those
associated with organ or tissue rejection or graft versus host
disease. Transplant antigens may be obtained or derived from cells
of a biological material or from information related to a
transplantable graft. Transplant antigens generally include
proteins, polypeptides, peptides, lipoproteins, glycolipids,
polynucleotides or are contained or expressed in cells. Information
related to a transplantable graft is any information about a
transplantable graft that can be used to obtain or derive
transplant antigens. Such information includes information about
antigens that would be expected to be present in or on cells of a
transplantable graft such as, for example, sequence information,
types or classes of antigens and/or their MHC Class I, MHC Class II
or B cell presentation restrictions. Such information may also
include information about the type of transplantable graft (e.g.,
autograft, allograft, xenograft), the molecular and cellular
composition of the graft, the bodily location from which the graft
is derived or to which the graft to be transplanted (e.g., whole or
partial organ, skin, bone, nerves, tendon, neurons, blood vessels,
fat, cornea, etc.).
[0073] Antigens also include antigens associated with a therapeutic
protein that can be presented for recognition by cells of the
immune system and that can generate an undesired immune response
against the therapeutic protein. Therapeutic protein antigens
generally include proteins, polypeptides, peptides, lipoproteins,
or are contained or expressed in, by or on cells.
[0074] Antigens, can be antigens that are fully defined or
characterized. However, in some embodiments, an antigen is not
fully defined or characterized. Antigens, therefore, also include
those that are contained within a cell or tissue preparation, cell
debris, cell exosome or conditioned media and can be delivered in
such form in some embodiments.
[0075] "Assessing an immune response" refers to any measurement or
determination of the level, presence or absence, reduction,
increase in, etc. of an immune response in vitro or in vivo. Such
measurements or determinations may be performed on one or more
samples obtained from a subject. Such assessing can be performed
with any of the methods provided herein or otherwise known in the
art.
[0076] An "at risk" subject is one in which a health practitioner
believes has a chance of having a disease, disorder or condition as
provided herein or is one a health practitioner believes has a
chance of experiencing an undesired immune response as provided
herein.
[0077] An "autoimmune disease" is any disease where the immune
system mounts an undesired immune response against self (e.g., one
or more autoantigens). In some embodiments, an autoimmune disease
comprises an aberrant destruction of cells of the body as part of
the self-targeted immune response. In some embodiments, the
destruction of self manifests in the malfunction of an organ, for
example, the colon or pancreas. Examples of autoimmune diseases are
described elsewhere herein. Additional autoimmune diseases will be
known to those of skill in the art and the invention is not limited
in this respect.
[0078] "Average", as used herein, refers to the arithmetic mean
unless otherwise noted.
[0079] "B cell antigen" means any antigen that triggers an immune
response in a B cell (e.g., an antigen that is specifically
recognized by a B cell or a receptor thereon). In some embodiments,
an antigen that is a T cell antigen is also a B cell antigen. In
other embodiments, the T cell antigen is not also a B cell antigen.
B cell antigens include, but are not limited to proteins, peptides,
small molecules, and carbohydrates. In some embodiments, the B cell
antigen comprises a non-protein antigen (i.e., not a protein or
peptide antigen). In some embodiments, the B cell antigen comprises
a autoantigen. In other embodiments, the B cell antigen is obtained
or derived from an allergen, autoantigen, therapeutic protein, or
transplantable graft.
[0080] "Concomitantly" means administering two or more substances
to a subject in a manner that is correlated in time, preferably
sufficiently correlated in time so as to provide a modulation in an
immune response. In embodiments, concomitant administration may
occur through administration of two or more substances in the same
dosage form. In other embodiments, concomitant administration may
encompass administration of two or more substances in different
dosage forms, but within a specified period of time, preferably
within 1 month, more preferably within 1 week, still more
preferably within 1 day, and even more preferably within 1
hour.
[0081] "Couple" or "Coupled" or "Couples" (and the like) means to
chemically associate one entity (for example a moiety) with
another. In some embodiments, the coupling is covalent, meaning
that the coupling occurs in the context of the presence of a
covalent bond between the two entities. In non-covalent
embodiments, the non-covalent coupling is mediated by non-covalent
interactions including but not limited to charge interactions,
affinity interactions, metal coordination, physical adsorption,
host-guest interactions, hydrophobic interactions, TT stacking
interactions, hydrogen bonding interactions, van der Waals
interactions, magnetic interactions, electrostatic interactions,
dipole-dipole interactions, and/or combinations thereof. In
embodiments, encapsulation is a form of coupling.
[0082] "Derived" means prepared from a material or information
related to a material but is not "obtained" from the material. Such
materials may be substantially modified or processed forms of
materials taken directly from a biological material. Such materials
also include materials produced from information related to a
biological material.
[0083] "Dosage form" means a pharmacologically and/or
immunologically active material in a medium, carrier, vehicle, or
device suitable for administration to a subject.
[0084] "Encapsulate" means to enclose at least a portion of a
substance within a synthetic nanocarrier. In some embodiments, a
substance is enclosed completely within a synthetic nanocarrier. In
other embodiments, most or all of a substance that is encapsulated
is not exposed to the local environment external to the synthetic
nanocarrier. In other embodiments, no more than 50%, 40%, 30%, 20%,
10% or 5% (weight/weight) is exposed to the local environment.
Encapsulation is distinct from absorption, which places most or all
of a substance on a surface of a synthetic nanocarrier, and leaves
the substance exposed to the local environment external to the
synthetic nanocarrier.
[0085] "Epitope", also known as an antigenic determinant, is the
part of an antigen that is recognized by the immune system,
specifically by, for example, antibodies, B cells, or T cells. As
used herein, "MHC Class I-restricted epitopes" are epitopes that
are presented to immune cells by MHC class I molecules found on
nucleated cells. "MHC Class II-restricted epitopes" are epitopes
that are presented to immune cells by MHC class II molecules found
on antigen-presenting cells (APCs), for example, on professional
antigen-presenting immune cells, such as on macrophages, B cells,
and dendritic cells, or on non-hematopoietic cells, such as
hepatocytes. "B cell epitopes" are molecular structures that are
recognized by antibodies or B cells. In some embodiments, the
epitope itself is an antigen.
[0086] A number of epitopes are known to those of skill in the art,
and exemplary epitopes suitable according to some aspects of this
invention include, but are not limited to those listed in the
Immune Epitope Database (www.immuneepitope.org, Vita R, Zarebski L,
Greenbaum J A, Emami H, Hoof I, Salimi N, Damle R, Sette A, Peters
B. The immune epitope database 2.0. Nucleic Acids Res. 2010
January; 38(Database issue):D854-62; the entire contents of which
as well as all database entries of IEDB version 2.4, August 2011,
and particularly all epitopes disclosed therein, are incorporated
herein by reference). Epitopes can also be identified with publicly
available algorithms, for example, the algorithms described in Wang
P, Sidney J, Kim Y, Sette A, Lund O, Nielsen M, Peters B. 2010.
peptide binding predictions for HLA DR, DP and DQ molecules. BMC
Bioinformatics 2010, 11:568; Wang P, Sidney J, Dow C, Mothe B,
Sette A, Peters B. 2008. A systematic assessment of MHC class II
peptide binding predictions and evaluation of a consensus approach.
PLoS Comput Biol. 4(4):e1000048; Nielsen M, Lund O. 2009. NN-align.
An artificial neural network-based alignment algorithm for MHC
class II peptide binding prediction. BMC Bioinformatics. 10:296;
Nielsen M, Lundegaard C, Lund O. 2007. Prediction of MHC class II
binding affinity using SMM-align, a novel stabilization matrix
alignment method. BMC Bioinformatics. 8:238; Bui H H, Sidney J,
Peters B, Sathiamurthy M, Sinichi A, Purton K A, Mothe B R, Chisari
F V, Watkins D I, Sette A. 2005. Immunogenetics. 57:304-314;
Sturniolo T, Bono E, Ding J, Raddrizzani L, Tuereci O, Sahin U,
Braxenthaler M, Gallazzi F, Protti M P, Sinigaglia F, Hammer J.
1999. Generation of tissue-specific and promiscuous HLA ligand
databases using DNA microarrays and virtual HLA class II matrices.
Nat Biotechnol. 17(6):555-561; Nielsen M, Lundegaard C, Worning P,
Lauemoller S L, Lamberth K, Buus S, Brunak S, Lund O. 2003.
Reliable prediction of T-cell epitopes using neural networks with
novel sequence representations. Protein Sci 12:1007-1017; Bui H H,
Sidney J, Peters B, Sathiamurthy M, Sinichi A, Purton K A, Mothe B
R, Chisari F V, Watkins D I, Sette A. 2005. Automated generation
and evaluation of specific MHC binding predictive tools: ARB matrix
applications. Immunogenetics 57:304-314; Peters B, Sette A. 2005.
Generating quantitative models describing the sequence specificity
of biological processes with the stabilized matrix method. BMC
Bioinformatics 6:132; Chou P Y, Fasman G D. 1978. Prediction of the
secondary structure of proteins from their amino acid sequence. Adv
Enzymol Relat Areas Mol Biol 47:45-148; Emini E A, Hughes J V,
Perlow D S, Boger J. 1985. Induction of hepatitis A
virus-neutralizing antibody by a virus-specific synthetic peptide.
J Virol 55:836-839; Karplus P A, Schulz G E. 1985. Prediction of
chain flexibility in proteins. Naturwissenschaften 72:212-213;
Kolaskar A S, Tongaonkar P C. 1990. A semi-empirical method for
prediction of antigenic determinants on protein antigens. FEBS Lett
276:172-174; Parker J M, Guo D, Hodges R S. 1986. New
hydrophilicity scale derived from high-performance liquid
chromatography peptide retention data: correlation of predicted
surface residues with antigenicity and X-ray-derived accessible
sites. Biochemistry 25:5425-5432; Larsen J E, Lund O, Nielsen M.
2006. Improved method for predicting linear B-cell epitopes.
Immunome Res 2:2; Ponomarenko J V, Bourne P E. 2007.
Antibody-protein interactions: benchmark datasets and prediction
tools evaluation. BMC Struct Biol 7:64; Haste Andersen P, Nielsen
M, Lund O. 2006. Prediction of residues in discontinuous B-cell
epitopes using protein 3D structures. Protein Sci 15:2558-2567;
Ponomarenko J V, Bui H, Li W, Fusseder N, Bourne P E, Sette A,
Peters B. 2008. ElliPro: a new structure-based tool for the
prediction of antibody epitopes. BMC Bioinformatics 9:514; Nielsen
M, Lundegaard C, Blicher T, Peters B, Sette A, Justesen S, Buus S,
and Lund O. 2008. PLoS Comput Biol. 4(7)e1000107. Quantitative
predictions of peptide binding to any HLA-DR molecule of known
sequence: NetMHCIIpan; the entire contents of each of which are
incorporated herein by reference for disclosure of methods and
algorithms for the identification of epitopes.
[0087] Other examples of epitopes include any of the MHC Class
I-restricted, MHC Class II-restricted and B cell epitopes as
provided as SEQ ID NOs: 1-943. Without wishing to being bound by
any particular theory, MHC Class I-restricted epitopes include
those set forth in SEQ ID NOs: 1-186, MHC Class II-restricted
epitopes include those set forth in SEQ ID NOs: 187-537, and B cell
epitopes include those set forth in SEQ ID NOs: 538-943. These
epitopes include MHC Class I-restricted autoantigens, MHC Class
II-restricted epitopes of allergens and B cell epitopes of
autoantigens and allergens.
[0088] "Generating" means causing an action, such as an immune
response (e.g., a tolerogenic immune response) to occur, either
directly oneself or indirectly, such as, but not limited to, an
unrelated third party that takes an action through reliance on
one's words or deeds.
[0089] "Identifying" is any action or set of actions that allows a
clinician to recognize a subject as one who may benefit from the
methods and compositions provided herein. Preferably, the
identified subject is one who is in need of a tolerogenic immune
response as provided herein. The action or set of actions may be
either directly oneself or indirectly, such as, but not limited to,
an unrelated third party that takes an action through reliance on
one's words or deeds.
[0090] "Immunosuppressant" means a compound that causes an APC to
have an immunosuppressive (e.g., tolerogenic effect). An
immunosuppressive effect generally refers to the production or
expression of cytokines or other factors by the APC that reduces,
inhibits or prevents an undesired immune response but can also
include the production or expression of cytokines or other factors
by the APC that increases, stimulates or promotes a desired immune
response. When the APC results in an immunosuppressive effect on
immune cells that recognize an antigen presented by the APC, the
immunosuppressive effect is said to be specific to the presented
antigen. Such effect is also referred to herein as a tolerogenic
effect. Without being bound by any particular theory, it is thought
that the immunosuppressive or tolerogenic effect is a result of the
immunosuppressant being delivered to the APC, preferably in the
presence of an antigen (e.g., an administered antigen or one that
is already present in vivo). Accordingly, the immunosuppressant
includes compounds that provide a tolerogenic immune response to an
antigen that may or may not be provided in the same composition or
a different composition. In one embodiment, the immunosuppressant
is one that causes an APC to promote a regulatory phenotype in one
or more immune effector cells. For example, the regulatory
phenotype may be characterized by the production, induction,
stimulation or recruitment of Treg cells, etc. This may be the
result of the conversion of B cells to a regulatory phenotype. This
may also be the result of the stimulation, recruitment, etc. of
regulatory B cells. This may also be the result of induction of
FoxP3 in other immune cells, such as macrophages and iNKT cells. In
one embodiment, the immunosuppressant is one that affects the
response of the APC after it processes an antigen. In another
embodiment, the immunosuppressant is not one that interferes with
the processing of the antigen. In a further embodiment, the
immunosuppressant is not an apoptotic-signaling molecule. In
another embodiment, the immunosuppressant is not a
phospholipid.
[0091] Immunosuppressants include, but are not limited to, statins;
mTOR inhibitors, such as rapamycin or a rapamycin analog;
TGF-.beta. signaling agents; TGF-.beta. receptor agonists; histone
deacetylase inhibitors, such as Trichostatin A; corticosteroids;
inhibitors of mitochondrial function, such as rotenone; P38
inhibitors; NF-.kappa..beta. inhibitors, such as 6Bio,
Dexamethasone, TCPA-1, IKK VII; adenosine receptor agonists;
prostaglandin E2 agonists (PGE2), such as Misoprostol;
phosphodiesterase inhibitors, such as phosphodiesterase 4 inhibitor
(PDE4), such as Rolipram; proteasome inhibitors; kinase inhibitors;
G-protein coupled receptor agonists; G-protein coupled receptor
antagonists; glucocorticoids; retinoids; cytokine inhibitors;
cytokine receptor inhibitors; cytokine receptor activators;
peroxisome proliferator-activated receptor antagonists; peroxisome
proliferator-activated receptor agonists; histone deacetylase
inhibitors; calcineurin inhibitors; phosphatase inhibitors; PI3 KB
inhibitors, such as TGX-221; autophagy inhibitors, such as
3-Methyladenine; aryl hydrocarbon receptor inhibitors; proteasome
inhibitor I (PSI); and oxidized ATPs, such as P2X receptor
blockers. Immunosuppressants also include IDO, vitamin D3,
cyclosporins, such as cyclosporine A, aryl hydrocarbon receptor
inhibitors, resveratrol, azathiopurine (Aza), 6-mercaptopurine
(6-MP), 6-thioguanine (6-TG), FK506, sanglifehrin A, salmeterol,
mycophenolate mofetil (MMF), aspirin and other COX inhibitors,
niflumic acid, estriol and triptolide. In embodiments, the
immunosuppressant may comprise any of the agents provided
herein.
[0092] The immunosuppressant can be a compound that directly
provides the immunosuppressive (e.g., tolerogenic) effect on APCs
or it can be a compound that provides the immunosuppressive (e.g.,
tolerogenic) effect indirectly (i.e., after being processed in some
way after administration). Immunosuppressants, therefore, include
prodrug forms of any of the compounds provided herein.
[0093] Immunosuppressants also include nucleic acids that encode
the peptides, polypeptides or proteins provided herein that result
in an immunosuppressive (e.g., tolerogenic) immune response. In
embodiments, therefore, the immunosuppressant is a nucleic acid
that encodes a peptide, polypeptide or protein that results in an
immunosuppressive (e.g., tolerogenic) immune response, and it is
the nucleic acid that is coupled to the synthetic nanocarrier.
[0094] The nucleic acid may be DNA or RNA, such as mRNA. In
embodiments, the inventive compositions comprise a complement, such
as a full-length complement, or a degenerate (due to degeneracy of
the genetic code) of any of the nucleic acids provided herein. In
embodiments, the nucleic acid is an expression vector that can be
transcribed when transfected into a cell line. In embodiments, the
expression vector may comprise a plasmid, retrovirus, or an
adenovirus amongst others. Nucleic acids can be isolated or
synthesized using standard molecular biology approaches, for
example by using a polymerase chain reaction to produce a nucleic
acid fragment, which is then purified and cloned into an expression
vector. Additional techniques useful in the practice of this
invention may be found in Current Protocols in Molecular Biology
2007 by John Wiley and Sons, Inc.; Molecular Cloning: A Laboratory
Manual (Third Edition) Joseph Sambrook, Peter MacCallum Cancer
Institute, Melbourne, Australia; David Russell, University of Texas
Southwestern Medical Center, Dallas, Cold Spring Harbor.
[0095] In embodiments, the immunosuppressants provided herein are
coupled to synthetic nanocarriers. In preferable embodiments, the
immunosuppressant is an element that is in addition to the material
that makes up the structure of the synthetic nanocarrier. For
example, in one embodiment, where the synthetic nanocarrier is made
up of one or more polymers, the immunosuppressant is a compound
that is in addition and coupled to the one or more polymers. As
another example, in one embodiment, where the synthetic nanocarrier
is made up of one or more lipids, the immunosuppressant is again in
addition and coupled to the one or more lipids. In embodiments,
such as where the material of the synthetic nanocarrier also
results in an immunosuppressive (e.g., tolerogenic) effect, the
immunosuppressant is an element present in addition to the material
of the synthetic nanocarrier that results in an immunosuppressive
(e.g., tolerogenic) effect.
[0096] Other exemplary immunosuppressants include, but are not
limited, small molecule drugs, natural products, antibodies (e.g.,
antibodies against CD20, CD3, CD4), biologics-based drugs,
carbohydrate-based drugs, nanoparticles, liposomes, RNAi, antisense
nucleic acids, aptamers, methotrexate, NSAIDs; fingolimod;
natalizumab; alemtuzumab; anti-CD3; tacrolimus (FK506), etc.
Further immunosuppressants, are known to those of skill in the art,
and the invention is not limited in this respect.
[0097] "Inflammatory disease" means any disease, disorder or
condition in which undesired inflammation occurs.
[0098] "Load" of the immunosuppressant or antigen is the amount of
the immunosuppressant or antigen coupled to a synthetic nanocarrier
based on the total weight of materials in an entire synthetic
nanocarrier (weight/weight). Generally, the load is calculated as
an average across a population of synthetic nanocarriers. In one
embodiment, the load of the immunosuppressant on average across the
first population of synthetic nanocarriers is between 0.0001% and
50%. In another embodiment, the load of the antigen on average
across the first and/or second population of synthetic nanocarriers
is between 0.0001% and 50%. In yet another embodiment, the load of
the immunosuppressant and/or antigen is between 0.01% and 20%. In a
further embodiment, the load of the immunosuppressant and/or
antigen is between 0.1% and 10%. In still a further embodiment, the
load of the immunosuppressant and/or antigen is between 1% and 10%.
In yet another embodiment, the load of the immunosuppressant and/or
the antigen is at least 0.1%, at least 0.2%, at least 0.3%, at
least 0.4%, at least 0.5%, at least 0.6%, at least 0.7%, at least
0.8%, at least 0.9%, at least 1%, at least 2%, at least 3%, at
least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at
least 9%, at least 10%, at least 11%, at least 12%, at least 13%,
at least 14%, at least 15%, at least 16%, at least 17%, at least
18%, at least 19% or at least 20% on average across a population of
synthetic nanocarriers. In yet a further embodiment, the load of
the immunosuppressant and/or the antigen is 0.1%, 0.2%, 0.3%, 0.4%,
0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%,
10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% on average
across a population of synthetic nanocarriers. In some embodiments
of the above embodiments, the load of the immunosuppressant and/or
the antigen is no more than 25% on average across a population of
synthetic nanocarriers. In embodiments, the load is calculated as
described in the Examples.
[0099] In embodiments of any of the compositions and methods
provided, the load may be calculated as follows: Approximately 3 mg
of synthetic nanocarriers are collected and centrifuged to separate
supernatant from synthetic nanocarrier pellet. Acetonitrile is
added to the pellet, and the sample is sonicated and centrifuged to
remove any insoluble material. The supernatant and pellet are
injected on RP-HPLC and absorbance is read at 278 nm. The .mu.g
found in the pellet is used to calculate % entrapped (load), .mu.g
in supernatant and pellet are used to calculate total .mu.g
recovered.
[0100] "Maintenance dose" refers to a dose that is administered to
a subject, after an initial dose has resulted in an
immunosuppressive (e.g., tolerogenic) response in a subject, to
sustain a desired immunosuppressive (e.g., tolerogenic) response. A
maintenance dose, for example, can be one that maintains the
tolerogenic effect achieved after the initial dose, prevents an
undesired immune response in the subject, or prevents the subject
becoming a subject at risk of experiencing an undesired immune
response, including an undesired level of an immune response. In
some embodiments, the maintenance dose is one that is sufficient to
sustain an appropriate level of a desired immune response. In some
embodiments, the maintenance dose is one that is sufficient to
sustain an appropriate antibody titre or level of regulatory B cell
number and/or activity necessary to sustain a tolerogenic immune
response or defend against a challenge with an agent that results
in an undesired immune response.
[0101] "Maximum dimension of a synthetic nanocarrier" means the
largest dimension of a nanocarrier measured along any axis of the
synthetic nanocarrier. "Minimum dimension of a synthetic
nanocarrier" means the smallest dimension of a synthetic
nanocarrier measured along any axis of the synthetic nanocarrier.
For example, for a spheroidal synthetic nanocarrier, the maximum
and minimum dimension of a synthetic nanocarrier would be
substantially identical, and would be the size of its diameter.
Similarly, for a cuboidal synthetic nanocarrier, the minimum
dimension of a synthetic nanocarrier would be the smallest of its
height, width or length, while the maximum dimension of a synthetic
nanocarrier would be the largest of its height, width or length. In
an embodiment, a minimum dimension of at least 75%, preferably at
least 80%, more preferably at least 90%, of the synthetic
nanocarriers in a sample, based on the total number of synthetic
nanocarriers in the sample, is equal to or greater than 100 nm. In
an embodiment, a maximum dimension of at least 75%, preferably at
least 80%, more preferably at least 90%, of the synthetic
nanocarriers in a sample, based on the total number of synthetic
nanocarriers in the sample, is equal to or less than 5 .mu.m.
Preferably, a minimum dimension of at least 75%, preferably at
least 80%, more preferably at least 90%, of the synthetic
nanocarriers in a sample, based on the total number of synthetic
nanocarriers in the sample, is greater than 110 nm, more preferably
greater than 120 nm, more preferably greater than 130 nm, and more
preferably still greater than 150 nm. Aspects ratios of the maximum
and minimum dimensions of inventive synthetic nanocarriers may vary
depending on the embodiment. For instance, aspect ratios of the
maximum to minimum dimensions of the synthetic nanocarriers may
vary from 1:1 to 1,000,000:1, preferably from 1:1 to 100,000:1,
more preferably from 1:1 to 10,000:1, more preferably from 1:1 to
1000:1, still more preferably from 1:1 to 100:1, and yet more
preferably from 1:1 to 10:1. Preferably, a maximum dimension of at
least 75%, preferably at least 80%, more preferably at least 90%,
of the synthetic nanocarriers in a sample, based on the total
number of synthetic nanocarriers in the sample is equal to or less
than 3 .mu.m, more preferably equal to or less than 2 .mu.m, more
preferably equal to or less than 1 .mu.m, more preferably equal to
or less than 800 nm, more preferably equal to or less than 600 nm,
and more preferably still equal to or less than 500 nm. In
preferred embodiments, a minimum dimension of at least 75%,
preferably at least 80%, more preferably at least 90%, of the
synthetic nanocarriers in a sample, based on the total number of
synthetic nanocarriers in the sample, is equal to or greater than
100 nm, more preferably equal to or greater than 120 nm, more
preferably equal to or greater than 130 nm, more preferably equal
to or greater than 140 nm, and more preferably still equal to or
greater than 150 nm. Measurement of synthetic nanocarrier
dimensions (e.g., diameter) is obtained by suspending the synthetic
nanocarriers in a liquid (usually aqueous) media and using dynamic
light scattering (DLS) (e.g. using a Brookhaven ZetaPALS
instrument). For example, a suspension of synthetic nanocarriers
can be diluted from an aqueous buffer into purified water to
achieve a final synthetic nanocarrier suspension concentration of
approximately 0.01 to 0.1 mg/mL. The diluted suspension may be
prepared directly inside, or transferred to, a suitable cuvette for
DLS analysis. The cuvette may then be placed in the DLS, allowed to
equilibrate to the controlled temperature, and then scanned for
sufficient time to acquire a stable and reproducible distribution
based on appropriate inputs for viscosity of the medium and
refractive indicies of the sample. The effective diameter, or mean
of the distribution, is then reported. "Dimension" or "size" or
"diameter" of synthetic nanocarriers means the mean of a particle
size distribution obtained using dynamic light scattering.
[0102] "MHC" refers to major histocompatibility complex, a large
genomic region or gene family found in most vertebrates that
encodes MHC molecules that display fragments or epitopes of
processed proteins on the cell surface. The presentation of
MHC:peptide on cell surfaces allows for surveillance by immune
cells, usually a T cell. There are two general classes of MHC
molecules: Class I and Class II. Generally, Class I MHC molecules
are found on nucleated cells and present peptides to cytotoxic T
cells. Class II MHC molecules are found on certain immune cells,
chiefly macrophages, B cells and dendritic cells, collectively
known as professional APCs. The best-known genes in the MHC region
are the subset that encodes antigen-presenting proteins on the cell
surface. In humans, these genes are referred to as human leukocyte
antigen (HLA) genes.
[0103] "Non-methoxy-terminated polymer" means a polymer that has at
least one terminus that ends with a moiety other than methoxy. In
some embodiments, the polymer has at least two termini that ends
with a moiety other than methoxy. In other embodiments, the polymer
has no termini that ends with methoxy. "Non-methoxy-terminated,
pluronic polymer" means a polymer other than a linear pluronic
polymer with methoxy at both termini. Polymeric nanoparticles as
provided herein can comprise non-methoxy-terminated polymers or
non-methoxy-terminated, pluronic polymers.
[0104] "Obtained" means taken directly from a material and used
with substantially no modification and/or processing.
[0105] "Pharmaceutically acceptable excipient" means a
pharmacologically inactive material used together with the recited
synthetic nanocarriers to formulate the inventive compositions.
Pharmaceutically acceptable excipients comprise a variety of
materials known in the art, including but not limited to
saccharides (such as glucose, lactose, and the like), preservatives
such as antimicrobial agents, reconstitution aids, colorants,
saline (such as phosphate buffered saline), and buffers.
[0106] "Protocol" refers to any dosing regimen of one or more
substances to a subject. A dosing regimen may include the amount,
frequency and/or mode of administration. In some embodiments, such
a protocol may be used to administer one or more compositions of
the invention to one or more test subjects. Immune responses in
these test subject can then be assessed to determine whether or not
the protocol was effective in reducing an undesired immune response
or generating a desired immune response (e.g., the promotion of a
tolerogenic effect). Any other therapeutic and/or prophylactic
effect may also be assessed instead of or in addition to the
aforementioned immune responses. Whether or not a protocol had a
desired effect can be determined using any of the methods provided
herein or otherwise known in the art. For example, a population of
cells may be obtained from a subject to which a composition
provided herein has been administered according to a specific
protocol in order to determine whether or not specific immune
cells, cytokines, antibodies, etc. were reduced, generated,
activated, etc. Useful methods for detecting the presence and/or
number of immune cells include, but are not limited to, flow
cytometric methods (e.g., FACS) and immunohistochemistry methods.
Antibodies and other binding agents for specific staining of immune
cell markers, are commercially available. Such kits typically
include staining reagents for multiple antigens that allow for
FACS-based detection, separation and/or quantitation of a desired
cell population from a heterogeneous population of cells.
[0107] "Providing a subject" is any action or set of actions that
causes a clinician to come in contact with a subject and administer
a composition provided herein thereto or to perform a method
provided herein thereupon. Preferably, the subject is one who is in
need of a tolerogenic immune response as provided herein. The
action or set of actions may be either directly oneself or
indirectly, such as, but not limited to, an unrelated third party
that takes an action through reliance on one's words or deeds.
[0108] "Regulatory B cells" are art-recognized and refer to a type
of tolerogenic B cells or that have suppressive regulatory
function. Surface markers and chemokine profiles characteristic of
regulatory B cells, as well as subsets of regulatory B cells (e.g.,
IL-10 producing Bregs) are known to those of skill in the art
(e.g., as described in DiLillo et al., B10 cells and regulatory B
cells balance immune response during inflammation, autoimmunity,
and cancer, Ann N. Y. Acad. Sci. 1183 (2010) 38-57, ISSN 0077-8923;
the entire contents of which are incorporated herein by reference).
The presence of regulatory B cells can be determined by
intracellular staining for IL-10 by flow cytometry. For example,
after treatment B cells can be stained for surface markers, then
fixed and permeabilized and stained for intracellular IL-10 and
analyzed by flow cytometry.
[0109] "Subject" means animals, including warm blooded mammals such
as humans and primates; avians; domestic household or farm animals
such as cats, dogs, sheep, goats, cattle, horses and pigs;
laboratory animals such as mice, rats and guinea pigs; fish;
reptiles; zoo and wild animals; and the like.
[0110] "Substantially no MHC Class I-restricted epitopes" refers to
the absence of MHC Class I-restricted epitopes in an amount that by
itself, within the context of the antigen, in conjunction with a
carrier or in conjunction with an inventive composition stimulates
substantial activation of an immune response specific to the
epitope presented in the Class I context. In embodiments, a
composition with substantially no MHC I-restricted epitopes does
not contain a measurable amount of MHC Class I-restricted epitopes
of an antigen. In other embodiments, such a composition may
comprise a measurable amount of MHC Class I-restricted epitopes of
an antigen but said amount is not effective to generate a
measurable cytotoxic T cell immune response (by itself, within the
context of the antigen, in conjunction with a carrier or in
conjunction with an inventive composition) or is not effective to
generate a significant measurable cytotoxic T cell immune response
(by itself, within the context of the antigen, in conjunction with
a carrier or in conjunction with an inventive composition). In some
embodiments, a significant measurable cytotoxic T cell immune
response is one that produces or would be expected to produce an
adverse clinical result in a subject. In other embodiments, a
significant measurable cytotoxic T cell immune response is one that
is greater than the level of the same type of immune response
produced by a control antigen (e.g., one known not to comprise MHC
Class I-restricted epitopes or to stimulate cytotoxic T cell immune
responses).
[0111] In embodiments, the compositions do not comprise MHC Class
I-restricted epitopes (by itself, within the context of the
antigen, in conjunction with a carrier or in conjunction with an
inventive composition) that generate antigen-specific cytotoxic T
cell immune responses or an undesired level thereof. In some
embodiments, to ensure that a composition does not comprise such
epitopes, antigens are selected such that they do not comprise MHC
Class I-restricted epitopes for coupling to the synthetic
nanocarriers as provided herein. In other embodiments, to ensure
that a composition does not comprise such epitopes, synthetic
nanocarriers coupled to antigen are produced and tested for
cytotoxic T cell immune responses. The appropriate synthetic
nanocarriers may then be selected.
[0112] "Synthetic nanocarrier(s)" means a discrete object that is
not found in nature, and that possesses at least one dimension that
is less than or equal to 5 microns in size. Albumin nanoparticles
are generally included as synthetic nanocarriers, however in
certain embodiments the synthetic nanocarriers do not comprise
albumin nanoparticles. In embodiments, inventive synthetic
nanocarriers do not comprise chitosan. In other embodiments,
inventive synthetic nanocarriers are not lipid-based nanoparticles.
In further embodiments, inventive synthetic nanocarriers do not
comprise a phospholipid.
[0113] A synthetic nanocarrier can be, but is not limited to, one
or a plurality of lipid-based nanoparticles (also referred to
herein as lipid nanoparticles, i.e., nanoparticles where the
majority of the material that makes up their structure are lipids),
polymeric nanoparticles, metallic nanoparticles, surfactant-based
emulsions, dendrimers, buckyballs, nanowires, virus-like particles
(i.e., particles that are primarily made up of viral structural
proteins but that are not infectious or have low infectivity),
peptide or protein-based particles (also referred to herein as
protein particles, i.e., particles where the majority of the
material that makes up their structure are peptides or proteins)
(such as albumin nanoparticles) and/or nanoparticles that are
developed using a combination of nanomaterials such as
lipid-polymer nanoparticles. Synthetic nanocarriers may be a
variety of different shapes, including but not limited to
spheroidal, cuboidal, pyramidal, oblong, cylindrical, toroidal, and
the like. Synthetic nanocarriers according to the invention
comprise one or more surfaces. Exemplary synthetic nanocarriers
that can be adapted for use in the practice of the present
invention comprise: (1) the biodegradable nanoparticles disclosed
in U.S. Pat. No. 5,543,158 to Gref et al., (2) the polymeric
nanoparticles of Published US Patent Application 20060002852 to
Saltzman et al., (3) the lithographically constructed nanoparticles
of Published US Patent Application 20090028910 to DeSimone et al.,
(4) the disclosure of WO 2009/051837 to von Andrian et al., (5) the
nanoparticles disclosed in Published US Patent Application
2008/0145441 to Penades et al., (6) the protein nanoparticles
disclosed in Published US Patent Application 20090226525 to de los
Rios et al., (7) the virus-like particles disclosed in published US
Patent Application 20060222652 to Sebbel et al., (8) the nucleic
acid coupled virus-like particles disclosed in published US Patent
Application 20060251677 to Bachmann et al., (9) the virus-like
particles disclosed in WO2010047839A1 or WO2009106999A2, (10) the
nanoprecipitated nanoparticles disclosed in P. Paolicelli et al.,
"Surface-modified PLGA-based Nanoparticles that can Efficiently
Associate and Deliver Virus-like Particles" Nanomedicine.
5(6):843-853 (2010), or (11) apoptotic cells, apoptotic bodies or
the synthetic or semisynthetic mimics disclosed in U.S. Publication
2002/0086049. In embodiments, synthetic nanocarriers may possess an
aspect ratio greater than 1:1, 1:1.2, 1:1.5, 1:2, 1:3, 1:5, 1:7, or
greater than 1:10.
[0114] Synthetic nanocarriers according to the invention that have
a minimum dimension of equal to or less than about 100 nm,
preferably equal to or less than 100 nm, do not comprise a surface
with hydroxyl groups that activate complement or alternatively
comprise a surface that consists essentially of moieties that are
not hydroxyl groups that activate complement. In a preferred
embodiment, synthetic nanocarriers according to the invention that
have a minimum dimension of equal to or less than about 100 nm,
preferably equal to or less than 100 nm, do not comprise a surface
that substantially activates complement or alternatively comprise a
surface that consists essentially of moieties that do not
substantially activate complement. In a more preferred embodiment,
synthetic nanocarriers according to the invention that have a
minimum dimension of equal to or less than about 100 nm, preferably
equal to or less than 100 nm, do not comprise a surface that
activates complement or alternatively comprise a surface that
consists essentially of moieties that do not activate complement.
In embodiments, synthetic nanocarriers exclude virus-like
particles. In embodiments, synthetic nanocarriers may possess an
aspect ratio greater than 1:1, 1:1.2, 1:1.5, 1:2, 1:3, 1:5, 1:7, or
greater than 1:10.
[0115] "T cell antigen" means a CD4+ T-cell antigen or CD8+ cell
antigen. "CD4+ T-cell antigen" means any antigen that is recognized
by and triggers an immune response in a CD4+ T-cell e.g., an
antigen that is specifically recognized by a T-cell receptor on a
CD4+ T cell via presentation of the antigen or portion thereof
bound to a Class II major histocompatability complex molecule
(MHC). "CD8+ T cell antigen" means any antigen that is recognized
by and triggers an immune response in a CD8+ T-cell e.g., an
antigen that is specifically recognized by a T-cell receptor on a
CD8+ T cell via presentation of the antigen or portion thereof
bound to a Class I major histocompatability complex molecule (MHC).
In some embodiments, an antigen that is a T cell antigen is also a
B cell antigen. In other embodiments, the T cell antigen is not
also a B cell antigen. T cell antigens generally are proteins or
peptides.
[0116] A "therapeutic protein" refers to any protein or
protein-based therapy that may be administered to a subject and
have a therapeutic effect. Such therapies include protein
replacement and protein supplementation therapies. Such therapies
also include the administration of exogenous or foreign protein,
antibody therapies, and cell or cell-based therapies. Therapeutic
proteins include enzymes, enzyme cofactors, hormones, blood
clotting factors, cytokines, growth factors, monoclonal antibodies
and polyclonal antibodies. Examples of other therapeutic proteins
are provided elsewhere herein. Therapeutic proteins may be produced
in, on or by cells and may be obtained from such cells or
administered in the form of such cells. In embodiments, the
therapeutic protein is produced in, on or by mammalian cells,
insect cells, yeast cells, bacteria cells, plant cells, transgenic
animal cells, transgenic plant cells, etc. The therapeutic protein
may be recombinantly produced in such cells. The therapeutic
protein may be produced in, on or by a virally transformed cell.
The therapeutic protein may also be produced in, on or by
autologous cells that have been transfected, transduced or
otherwise manipulated to express it. Alternatively, the therapeutic
protein may be administered as a nucleic acid or by introducing a
nucleic acid into a virus, VLP, liposome, etc. Alternatively, the
therapeutic protein may be obtained from such forms and
administered as the therapeutic protein itself. Subjects,
therefore, include any subject that has received, is receiving or
will receive any of the foregoing. Such subject includes subjects
that have received, is receiving or will receive gene therapy,
autologous cells that have been transfected, transduced or
otherwise manipulated to express a therapeutic protein, polypeptide
or peptide; or cells that express a therapeutic protein,
polypeptide or peptide.
[0117] "Therapeutic protein antigen" means an antigen that is
associated with a therapeutic protein that can be, or a portion of
which can be, presented for recognition by cells of the immune
system and can generate an undesired immune response (e.g., the
production of therapeutic protein-specific antibodies) against the
therapeutic protein. Therapeutic protein antigens generally include
proteins, polypeptides, peptides, lipoproteins, or are contained or
expressed in, on or by cells.
[0118] "Tolerogenic immune response" means any immune response that
can lead to immune suppression specific to an antigen or a cell,
tissue, organ, etc. that expresses such an antigen. Such immune
responses include any reduction, delay or inhibition in an
undesired immune response specific to the antigen or cell, tissue,
organ, etc. that expresses such antigen. Such immune responses also
include any stimulation, production, induction, promotion or
recruitment in a desired immune response specific to the antigen or
cell, tissue, organ, etc. that expresses such antigen. Tolerogenic
immune responses, therefore, include the absence of or reduction in
an undesired immune response to an antigen that can be mediated by
antigen reactive cells as well as the presence or promotion of
suppressive cells. Tolerogenic immune responses as provided herein
include immunological tolerance. To "generate a tolerogenic immune
response" refers to the generation of any of the foregoing immune
responses specific to an antigen or cell, tissue, organ, etc. that
expresses such antigen. The tolerogenic immune response can be the
result of MHC Class I-restricted presentation and/or MHC Class
II-restricted presentation and/or B cell presentation and/or
presentation by CD1d, etc. In embodiments, to ensure that a
composition comprises desired epitopes, antigens are selected such
that they comprise such epitopes for coupling to the synthetic
nanocarriers as provided herein. In other embodiments, to ensure
that a composition comprises such epitopes, synthetic nanocarriers
coupled to an antigen are produced and tested for regulatory B cell
immune responses, such as activation or generation. The appropriate
synthetic nanocarriers may then be selected.
[0119] Tolerogenic immune responses include any reduction, delay or
inhibition in CD4+ T cell, CD8+ T cell or B cell proliferation
and/or activity. Tolerogenic immune responses also include a
reduction in antigen-specific antibody production. Tolerogenic
immune responses can also include any response that leads to the
stimulation, induction, production or recruitment of regulatory
cells, such as CD4+ Treg cells, CD8+ Treg cells, Breg cells, etc.
In some embodiments, the tolerogenic immune response, is one that
results in the conversion to a regulatory phenotype characterized
by the production, induction, stimulation or recruitment of
regulatory cells.
[0120] Tolerogenic immune responses also include any response that
leads to the stimulation, production or recruitment of CD4+ Treg
cells and/or CD8+ Treg cells. CD4+ Treg cells can express the
transcription factor FoxP3 and inhibit inflammatory responses and
auto-immune inflammatory diseases (Human regulatory T cells in
autoimmune diseases. Cvetanovich G L, Hafler D A. Curr Opin
Immunol. 2010 December; 22(6):753-60. Regulatory T cells and
autoimmunity. Vila J, Isaacs J D, Anderson A E. Curr Opin Hematol.
2009 July; 16(4):274-9). Such cells also suppress T-cell help to
B-cells and induce tolerance to both self and foreign antigens
(Therapeutic approaches to allergy and autoimmunity based on FoxP3+
regulatory T-cell activation and expansion. Miyara M, Wing K,
Sakaguchi S. J Allergy Clin Immunol. 2009 April; 123(4):749-55).
CD4+ Treg cells recognize antigen when presented by Class II
proteins on APCs. CD8+ Treg cells, which recognize antigen
presented by Class I (and Qa-1), can also suppress T-cell help to
B-cells and result in activation of antigen-specific suppression
inducing tolerance to both self and foreign antigens. Disruption of
the interaction of Qa-1 with CD8+ Treg cells has been shown to
dysregulate immune responses and results in the development of
auto-antibody formation and an auto-immune lethal
systemic-lupus-erythematosus (Kim et al., Nature. 2010 Sep. 16, 467
(7313): 328-32). CD8+ Treg cells have also been shown to inhibit
models of autoimmune inflammatory diseases including rheumatoid
arthritis and colitis (CD4+CD25+ regulatory T cells in autoimmune
arthritis. Oh S, Rankin A L, Caton A J. Immunol Rev. 2010 January;
233(1):97-111. Regulatory T cells in inflammatory bowel disease.
Boden E K, Snapper S B. Curr Opin Gastroenterol. 2008 November;
24(6):733-41). In some embodiments, the compositions provided can
effectively result in both types of responses (CD4+ Treg and CD8+
Treg). In other embodiments, FoxP3 can be induced in other immune
cells, such as macrophages, iNKT cells, etc., and the compositions
provided herein can result in one or more of these responses as
well.
[0121] Tolerogenic immune responses also include, but are not
limited to, the induction of regulatory cytokines, such as Treg
cytokines; induction of inhibitory cytokines; the inhibition of
inflammatory cytokines (e.g., IL-4, IL-1b, IL-5, TNF-.alpha., IL-6,
GM-CSF, IFN-.gamma., IL-2, IL-9, IL-12, IL-17, IL-18, IL-21, IL-22,
IL-23, M-CSF, C reactive protein, acute phase protein, chemokines
(e.g., MCP-1, RANTES, MIP-1.alpha., MIP-1.beta., MIG, ITAC or
IP-10), the production of anti-inflammatory cytokines (e.g., IL-4,
IL-13, IL-10, etc.), chemokines (e.g., CCL-2, CXCL8), proteases
(e.g., MMP-3, MMP-9), leukotrienes (e.g., CysLT-1, CysLT-2),
prostaglandins (e.g., PGE2) or histamines; the inhibition of
polarization to a Th17, Th1 or Th2 immune response; the inhibition
of effector cell-specific cytokines: Th17 (e.g., IL-17, IL-25), Th1
(IFN-.gamma.), Th2 (e.g., IL-4, IL-13); the inhibition of Th1-,
Th2- or TH17-specific transcription factors; the inhibition of
proliferation of effector T cells; the induction of apoptosis of
effector T cells; the induction of tolerogenic dendritic
cell-specific genes, the induction of FoxP3 expression, the
inhibition of IgE induction or IgE-mediated immune responses; the
inhibition of antibody responses (e.g., antigen-specific antibody
production); the inhibition of T helper cell response; the
production of TGF-.beta. and/or IL-10; the inhibition of effector
function of autoantibodies (e.g., inhibition in the depletion of
cells, cell or tissue damage or complement activation); etc.
[0122] Any of the foregoing may be measured in vivo in one or more
animal models or may be measured in vitro. One of ordinary skill in
the art is familiar with such in vivo or in vitro measurements.
Undesired immune responses or tolerogenic immune responses can be
monitored using, for example, methods of assessing immune cell
number and/or function, tetramer analysis, ELISPOT, flow
cytometry-based analysis of cytokine expression, cytokine
secretion, cytokine expression profiling, gene expression
profiling, protein expression profiling, analysis of cell surface
markers, PCR-based detection of immune cell receptor gene usage
(see T. Clay et al., "Assays for Monitoring Cellular Immune
Response to Active Immunotherapy of Cancer" Clinical Cancer
Research 7:1127-1135 (2001)), etc. Undesired immune responses or
tolerogenic immune responses may also be monitored using, for
example, methods of assessing protein levels in plasma or serum,
immune cell proliferation and/or functional assays, etc. In some
embodiments, tolerogenic immune responses can be monitored by
assessing the induction of FoxP3. In addition, specific methods are
described in more detail in the Examples.
[0123] Preferably, tolerogenic immune responses lead to the
inhibition of the development, progression or pathology of the
diseases, disorders or conditions described herein. Whether or not
the inventive compositions can lead to the inhibition of the
development, progression or pathology of the diseases, disorders or
conditions described herein can be measured with animal models of
such diseases, disorders or conditions. In some embodiments, the
reduction of an undesired immune response or generation of a
tolerogenic immune response may be assessed by determining clinical
endpoints, clinical efficacy, clinical symptoms, disease biomarkers
and/or clinical scores. Undesired immune responses or tolerogenic
immune responses can also be assessed with diagnostic tests to
assess the presence or absence of a disease, disorder or condition
as provided herein. Undesired or desired immune responses can
further be assessed by methods of measuring therapeutic proteins
levels and/or function in a subject. In embodiments, methods for
monitoring or assessing undesired allergic responses include
assessing an allergic response in a subject by skin reactivity
and/or allergen-specific antibody production.
[0124] In some embodiments, monitoring or assessing the generation
of an undesired immune response or a tolerogenic immune response in
a subject can be prior to the administration of a composition of
synthetic nanocarriers provided herein and/or prior to
administration of a transplantable graft or therapeutic protein or
exposure to an allergen. In other embodiments, assessing the
generation of an undesired immune response or tolerogenic immune
response can be after administration of a composition of synthetic
nanocarriers provided herein and/or after administration of a
transplantable graft or therapeutic protein or exposure to an
allergen. In some embodiments, the assessment is done after
administration of the composition of synthetic nanocarriers, but
prior to administration of a transplantable graft or therapeutic
protein or exposure to an allergen. In other embodiments, the
assessment is done after administration of a transplantable graft
or therapeutic protein or exposure to an allergen, but prior to
administration of the composition. In still other embodiments, the
assessment is performed prior to both the administration of the
synthetic nanocarriers and administration of a transplantable graft
or therapeutic protein or exposure to an allergen, while in yet
other embodiments the assessment is performed after both the
administration of synthetic nanocarriers and the administration of
a transplantable graft or therapeutic protein or exposure to an
allergen. In further embodiments, the assessment is performed both
prior to and after the administration of the synthetic nanocarriers
and/or administration of a transplantable graft or therapeutic
protein or exposure to an allergen. In still other embodiments, the
assessment is performed more than once on the subject to determine
that a desirable immune state is maintained in the subject, such as
a subject that has or is at risk of having an inflammatory disease,
an autoimmune disease, an allergy, organ or tissue rejection or
graft verus host disease. Other subjects include those that have
undergone or will undergo transplantation as well as those that
have received, are receiving or will receive a therapeutic protein
against which they have experienced, are experiencing or are
expected to experience an undesired immune response.
[0125] An antibody response can be assessed by determining one or
more antibody titers. "Antibody titer" means a measurable level of
antibody production. Methods for measuring antibody titers are
known in the art and include Enzyme-linked Immunosorbent Assay
(ELISA). In embodiments, the antibody response can be quantitated,
for example, as the number of antibodies, concentration of
antibodies or titer. The values can be absolute or they can be
relative. Assays for quantifying an antibody response include
antibody capture assays, enzyme-linked immunosorbent assays
(ELISAs), inhibition liquid phase absorption assays (ILPAAs),
rocket immunoelectrophoresis (RIE) assays and line
immunoelectrophoresis (LIE) assays. When an antibody response is
compared to another antibody response the same type of quantitative
value (e.g., titer) and method of measurement (e.g., ELISA) is
preferably used to make the comparison.
[0126] An ELISA method for measuring an antibody titer, for
example, a typical sandwich ELISA, may consist of the following
steps (i) preparing an ELISA-plate coating material such that the
antibody target of interest is coupled to a substrate polymer or
other suitable material (ii) preparing the coating material in an
aqueous solution (such as PBS) and delivering the coating material
solution to the wells of a multiwell plate for overnight deposition
of the coating onto the multiwell plate (iii) thoroughly washing
the multiwell plate with wash buffer (such as 0.05% Tween-20 in
PBS) to remove excess coating material (iv) blocking the plate for
nonspecific binding by applying a diluent solution (such as 10%
fetal bovine serum in PBS), (v) washing the blocking/diluent
solution from the plate with wash buffer (vi) diluting the serum
sample(s) containing antibodies and appropriate standards (positive
controls) with diluent as required to obtain a concentration that
suitably saturates the ELISA response (vii) serially diluting the
plasma samples on the multiwell plate such to cover a range of
concentrations suitable for generating an ELISA response curve
(viii) incubating the plate to provide for antibody-target binding
(ix) washing the plate with wash buffer to remove antibodies not
bound to antigen (x) adding an appropriate concentration of a
secondary detection antibody in same diluent such as a
biotin-coupled detection antibody capable of binding the primary
antibody (xi) incubating the plate with the applied detection
antibody, followed by washing with wash buffer (xii) adding an
enzyme such as streptavidin-HRP (horse radish peroxidase) that will
bind to biotin found on biotinylated antibodies and incubating
(xiii) washing the multiwell plate (xiv) adding substrate(s) (such
as TMB solution) to the plate (xv) applying a stop solution (such
as 2N sulfuric acid) when color development is complete (xvi)
reading optical density of the plate wells at a specific wavelength
for the substrate (450 nm with subtraction of readings at 570 nm)
(xvi) applying a suitable multiparameter curve fit to the data and
defining half-maximal effective concentration (EC50) as the
concentration on the curve at which half the maximum OD value for
the plate standards is achieved.
[0127] A "transplantable graft" refers to a biological material,
such as cells, tissues and organs (in whole or in part) that can be
administered to a subject. Transplantable grafts may be autografts,
allografts, or xenografts of, for example, a biological material
such as an organ, tissue, skin, bone, nerves, tendon, neurons,
blood vessels, fat, cornea, pluripotent cells, differentiated cells
(obtained or derived in vivo or in vitro), etc. In some
embodiments, a transplantable graft is formed, for example, from
cartilage, bone, extracellular matrix, or collagen matrices.
Transplantable grafts may also be single cells, suspensions of
cells and cells in tissues and organs that can be transplanted.
Transplantable cells typically have a therapeutic function, for
example, a function that is lacking or diminished in a recipient
subject. Some non-limiting examples of transplantable cells are
.beta.-cells, hepatocytes, hematopoietic stem cells, neuronal stem
cells, neurons, glial cells, or myelinating cells. Transplantable
cells can be cells that are unmodified, for example, cells obtained
from a donor subject and usable in transplantation without any
genetic or epigenetic modifications. In other embodiments,
transplantable cells can be modified cells, for example, cells
obtained from a subject having a genetic defect, in which the
genetic defect has been corrected, or cells that are derived from
reprogrammed cells, for example, differentiated cells derived from
cells obtained from a subject.
[0128] "Transplantation" refers to the process of transferring
(moving) a transplantable graft into a recipient subject (e.g.,
from a donor subject, from an in vitro source (e.g., differentiated
autologous or heterologous native or induced pluripotent cells))
and/or from one bodily location to another bodily location in the
same subject.
[0129] "Undesired immune response" refers to any undesired immune
response that results from exposure to an antigen, promotes or
exacerbates a disease, disorder or condition provided herein (or a
symptom thereof), or is symptomatic of a disease, disorder or
condition provided herein. Such immune responses generally have a
negative impact on a subject's health or is symptomatic of a
negative impact on a subject's health.
C. INVENTIVE COMPOSITIONS
[0130] Provided herein are methods and compositions for generating
regulatory B cells. Regulatory B cells are believed to modulate
immune responses, for example, in that IL-10 release from
regulatory B cells exerts anti-inflammatory and immunosuppressive
effects on most hematopoietic cells. IL-10 also suppresses
pro-inflammatory cytokine production by monocytes and macrophages
and the proliferation of antigen-specific CD4+ T cells. Without
wishing to be bound by any particular theory, it is believed that
the compositions provided herein can affect the number and/or
activity of regulatory B cells in vitro and/or in vivo by, for
example, interacting with naive B cells or regulatory B cell
precursors, resulting in an induction of regulatory B cell
maturation or an increase in regulatory B cell proliferation and/or
activity. The effect on regulatory B cell number and/or activity
may also be a result of the generation of other regulatory cell or
tolerogenic immune responses, such as through regulatory T cell
production and/or activation.
[0131] Provided herein are tolerogenic methods that include the
administration of synthetic nanocarrier compositions comprising
immunosuppressants and B cell and/or MHC Class II-restricted
epitopes of an antigen and related compositions. Such methods and
compositions are useful for generating, stimulating or recruiting
regulatory B cells and promoting the generation of tolerogenic
immune responses. The methods provided may also include the
administration therapeutic proteins and/or transplantable grafts.
The compositions provided, therefore, may also include the
therapeutic proteins and/or transplantable grafts. The compositions
may be administered to subjects in which a tolerogenic immune
response is desired. Such subjects include those that have or are
at risk of having an inflammatory disease, an autoimmune disease,
an allergy, organ or tissue rejection or graft versus host disease.
Such subjects also include those that have been, are being or will
be administered a therapeutic protein against which the subject has
experienced or is expected to experience an undesired immune
response. Such subjects also include those that have undergone or
will undergo transplantation.
[0132] Preferably, the compositions of the invention result in the
generation, recruitment or activation of regulatory B cells. One
hallmark of regulatory B cells is the ability to produce and
secrete IL-10. Some regulatory B cells are CD1d+ or CD1dhigh. Some
regulatory B cells are CD5+ and/or CD19+. For example, some
regulatory B cells are CD1dhighCD5+CD19+. Some regulatory B cells
are CD24+ or CD24high; and/or CD38+ or CD38high. For example, some
regulatory B cells are CD19+CD24highCD38high. Some regulatory B
cells produce and secrete IL-10. Additional surface markers and
chemokine secretion profiles that can be used to identify
regulatory B cells are known to those of skill in the art. Based on
the knowledge of surface markers useful for the identification of
various regulatory B cell populations, those of skill in the art
are able to identify and enumerate regulatory B cells in a
heterogeneous population of cells, for example, in a population of
cells in culture or in a population of cells obtained from a
subject.
[0133] As mentioned above, the synthetic nanocarriers are designed
to comprise B cell eptiopes and/or MHC Class II-restricted epitopes
and/or immunosuppressants. A wide variety of synthetic nanocarriers
can be used according to the invention. In some embodiments,
synthetic nanocarriers are spheres or spheroids. In some
embodiments, synthetic nanocarriers are flat or plate-shaped. In
some embodiments, synthetic nanocarriers are cubes or cubic. In
some embodiments, synthetic nanocarriers are ovals or ellipses. In
some embodiments, synthetic nanocarriers are cylinders, cones, or
pyramids.
[0134] In some embodiments, it is desirable to use a population of
synthetic nanocarriers that is relatively uniform in terms of size,
shape, and/or composition so that each synthetic nanocarrier has
similar properties. For example, at least 80%, at least 90%, or at
least 95% of the synthetic nanocarriers, based on the total number
of synthetic nanocarriers, may have a minimum dimension or maximum
dimension that falls within 5%, 10%, or 20% of the average diameter
or average dimension of the synthetic nanocarriers. In some
embodiments, a population of synthetic nanocarriers may be
heterogeneous with respect to size, shape, and/or composition.
[0135] Synthetic nanocarriers can be solid or hollow and can
comprise one or more layers. In some embodiments, each layer has a
unique composition and unique properties relative to the other
layer(s). To give but one example, synthetic nanocarriers may have
a core/shell structure, wherein the core is one layer (e.g. a
polymeric core) and the shell is a second layer (e.g. a lipid
bilayer or monolayer). Synthetic nanocarriers may comprise a
plurality of different layers.
[0136] In some embodiments, synthetic nanocarriers may optionally
comprise one or more lipids. In some embodiments, a synthetic
nanocarrier may comprise a liposome. In some embodiments, a
synthetic nanocarrier may comprise a lipid bilayer. In some
embodiments, a synthetic nanocarrier may comprise a lipid
monolayer. In some embodiments, a synthetic nanocarrier may
comprise a micelle. In some embodiments, a synthetic nanocarrier
may comprise a core comprising a polymeric matrix surrounded by a
lipid layer (e.g., lipid bilayer, lipid monolayer, etc.). In some
embodiments, a synthetic nanocarrier may comprise a non-polymeric
core (e.g., metal particle, quantum dot, ceramic particle, bone
particle, viral particle, proteins, nucleic acids, carbohydrates,
etc.) surrounded by a lipid layer (e.g., lipid bilayer, lipid
monolayer, etc.).
[0137] In other embodiments, synthetic nanocarriers may comprise
metal particles, quantum dots, ceramic particles, etc. In some
embodiments, a non-polymeric synthetic nanocarrier is an aggregate
of non-polymeric components, such as an aggregate of metal atoms
(e.g., gold atoms).
[0138] In some embodiments, synthetic nanocarriers may optionally
comprise one or more amphiphilic entities. In some embodiments, an
amphiphilic entity can promote the production of synthetic
nanocarriers with increased stability, improved uniformity, or
increased viscosity. In some embodiments, amphiphilic entities can
be associated with the interior surface of a lipid membrane (e.g.,
lipid bilayer, lipid monolayer, etc.). Many amphiphilic entities
known in the art are suitable for use in making synthetic
nanocarriers in accordance with the present invention. Such
amphiphilic entities include, but are not limited to,
phosphoglycerides; phosphatidylcholines; dipalmitoyl
phosphatidylcholine (DPPC); dioleylphosphatidyl ethanolamine
(DOPE); dioleyloxypropyltriethylammonium (DOTMA);
dioleoylphosphatidylcholine; cholesterol; cholesterol ester;
diacylglycerol; diacylglycerolsuccinate; diphosphatidyl glycerol
(DPPG); hexanedecanol; fatty alcohols such as polyethylene glycol
(PEG); polyoxyethylene-9-lauryl ether; a surface active fatty acid,
such as palmitic acid or oleic acid; fatty acids; fatty acid
monoglycerides; fatty acid diglycerides; fatty acid amides;
sorbitan trioleate (Span.RTM.85) glycocholate; sorbitan monolaurate
(Span.RTM.20); polysorbate 20 (Tween.RTM.20); polysorbate 60
(Tween.RTM.60); polysorbate 65 (Tween.RTM.65); polysorbate 80
(Tween.RTM.80); polysorbate 85 (Tween.RTM.85); polyoxyethylene
monostearate; surfactin; a poloxomer; a sorbitan fatty acid ester
such as sorbitan trioleate; lecithin; lysolecithin;
phosphatidylserine; phosphatidylinositol; sphingomyelin;
phosphatidylethanolamine (cephalin); cardiolipin; phosphatidic
acid; cerebrosides; dicetylphosphate;
dipalmitoylphosphatidylglycerol; stearylamine; dodecylamine;
hexadecyl-amine; acetyl palmitate; glycerol ricinoleate; hexadecyl
sterate; isopropyl myristate; tyloxapol; poly(ethylene
glycol)5000-phosphatidylethanolamine; poly(ethylene
glycol)400-monostearate; phospholipids; synthetic and/or natural
detergents having high surfactant properties; deoxycholates;
cyclodextrins; chaotropic salts; ion pairing agents; and
combinations thereof. An amphiphilic entity component may be a
mixture of different amphiphilic entities. Those skilled in the art
will recognize that this is an exemplary, not comprehensive, list
of substances with surfactant activity. Any amphiphilic entity may
be used in the production of synthetic nanocarriers to be used in
accordance with the present invention.
[0139] In some embodiments, synthetic nanocarriers may optionally
comprise one or more carbohydrates. Carbohydrates may be natural or
synthetic. A carbohydrate may be a derivatized natural
carbohydrate. In certain embodiments, a carbohydrate comprises
monosaccharide or disaccharide, including but not limited to
glucose, fructose, galactose, ribose, lactose, sucrose, maltose,
trehalose, cellbiose, mannose, xylose, arabinose, glucoronic acid,
galactoronic acid, mannuronic acid, glucosamine, galatosamine, and
neuramic acid. In certain embodiments, a carbohydrate is a
polysaccharide, including but not limited to pullulan, cellulose,
microcrystalline cellulose, hydroxypropyl methylcellulose (HPMC),
hydroxycellulose (HC), methylcellulose (MC), dextran, cyclodextran,
glycogen, hydroxyethylstarch, carageenan, glycon, amylose,
chitosan, N,O-carboxylmethylchitosan, algin and alginic acid,
starch, chitin, inulin, konjac, glucommannan, pustulan, heparin,
hyaluronic acid, curdlan, and xanthan. In embodiments, the
inventive synthetic nanocarriers do not comprise (or specifically
exclude) carbohydrates, such as a polysaccharide. In certain
embodiments, the carbohydrate may comprise a carbohydrate
derivative such as a sugar alcohol, including but not limited to
mannitol, sorbitol, xylitol, erythritol, maltitol, and
lactitol.
[0140] In some embodiments, synthetic nanocarriers can comprise one
or more polymers. In some embodiments, the synthetic nanocarriers
comprise one or more polymers that is a nonmethoxy-terminated,
pluronic polymer. In some embodiments, at least 1%, 2%, 3%, 4%, 5%,
10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,
75%, 80%, 85%, 90%, 95%, 97%, or 99% (weight/weight) of the
polymers that make up the synthetic nanocarriers are
non-methoxy-terminated, pluronic polymers. In some embodiments, all
of the polymers that make up the synthetic nanocarriers are
non-methoxy-terminated, pluronic polymers. In some embodiments, the
synthetic nanocarriers can comprise one or more polymers that is a
non-methoxy-terminated polymer. In some embodiments, at least 1%,
2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% (weight/weight)
of the polymers that make up the synthetic nanocarriers are
nonmethoxy-terminated polymers. In some embodiments, all of the
polymers that make up the synthetic nanocarriers are
non-methoxy-terminated polymers. In some embodiments, the synthetic
nanocarriers comprise one or more polymers that do not comprise
pluronic polymer. In some embodiments, at least 1%, 2%, 3%, 4%, 5%,
10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,
75%, 80%, 85%, 90%, 95%, 97%, or 99% (weight/weight) of the
polymers that make up the synthetic nanocarriers do not comprise
pluronic polymer. In some embodiments, all of the polymers that
make up the synthetic nanocarriers do not comprise pluronic
polymer. In some embodiments, such a polymer can be surrounded by a
coating layer (e.g., liposome, lipid monolayer, micelle, etc.). In
some embodiments, various elements of the synthetic nanocarriers
can be coupled with the polymer.
[0141] The immunosuppressants and/or antigens can be coupled to the
synthetic nanocarriers by any of a number of methods. Generally,
the coupling can be a result of bonding between the
immunosuppressants and/or antigens and the synthetic nanocarriers.
This bonding can result in the immunosuppressants and/or antigens
being attached to the surface of the synthetic nanocarrierss and/or
contained within (encapsulated) the synthetic nanocarriers. In some
embodiments, however, the immunosuppressants and/or antigens are
encapsulated by the synthetic nanocarriers as a result of the
structure of the synthetic nanocarriers rather than bonding to the
synthetic nanocarriers. In preferable embodiments, the synthetic
nanocarriers comprise a polymer as provided herein, and the
immunosuppressants and/or antigens are coupled to the polymer.
[0142] When coupling occurs as a result of bonding between the
immunosuppressants and/or antigens and synthetic nanocarriers, the
coupling may occur via a coupling moiety. A coupling moiety can be
any moiety through which an immunosuppressant and/or antigen is
bonded to a synthetic nanocarrier. Such moieties include covalent
bonds, such as an amide bond or ester bond, as well as separate
molecules that bond (covalently or non-covalently) the
immunosuppressant and/or antigen to the synthetic nanocarrier. Such
molecules include linkers or polymers or a unit thereof. For
example, the coupling moiety can comprise a charged polymer to
which an immunosuppressant and/or antigen electrostatically binds.
As another example, the coupling moiety can comprise a polymer or
unit thereof to which it is covalently bonded.
[0143] In preferred embodiments, the synthetic nanocarriers
comprise a polymer as provided herein. These synthetic nanocarriers
can be completely polymeric or they can be a mix of polymers and
other materials.
[0144] In some embodiments, the polymers of a synthetic nanocarrier
associate to form a polymeric matrix. In some of these embodiments,
a component, such as an immunosuppressant or antigen, can be
covalently associated with one or more polymers of the polymeric
matrix. In some embodiments, covalent association is mediated by a
linker. In some embodiments, a component can be noncovalently
associated with one or more polymers of the polymeric matrix. For
example, in some embodiments a component can be encapsulated
within, surrounded by, and/or dispersed throughout a polymeric
matrix. Alternatively or additionally, a component can be
associated with one or more polymers of a polymeric matrix by
hydrophobic interactions, charge interactions, van der Waals
forces, etc. A wide variety of polymers and methods for forming
polymeric matrices therefrom are known conventionally.
[0145] Polymers may be natural or unnatural (synthetic) polymers.
Polymers may be homopolymers or copolymers comprising two or more
monomers. In terms of sequence, copolymers may be random, block, or
comprise a combination of random and block sequences. Typically,
polymers in accordance with the present invention are organic
polymers.
[0146] In some embodiments, the polymer comprises a polyester,
polycarbonate, polyamide, or polyether, or unit thereof. In other
embodiments, the polymer comprises poly(ethylene glycol) (PEG),
polypropylene glycol, poly(lactic acid), poly(glycolic acid),
poly(lactic-co-glycolic acid), or a polycaprolactone, or unit
thereof. In some embodiments, it is preferred that the polymer is
biodegradable. Therefore, in these embodiments, it is preferred
that if the polymer comprises a polyether, such as poly(ethylene
glycol) or polypropylene glycol or unit thereof, the polymer
comprises a block-co-polymer of a polyether and a biodegradable
polymer such that the polymer is biodegradable. In other
embodiments, the polymer does not solely comprise a polyether or
unit thereof, such as poly(ethylene glycol) or polypropylene glycol
or unit thereof.
[0147] Other examples of polymers suitable for use in the present
invention include, but are not limited to polyethylenes,
polycarbonates (e.g. poly(1,3-dioxan-2one)), polyanhydrides (e.g.
poly(sebacic anhydride)), polypropylfumerates, polyamides (e.g.
polycaprolactam), polyacetals, polyethers, polyesters (e.g.,
polylactide, polyglycolide, polylactide-co-glycolide,
polycaprolactone, polyhydroxyacid (e.g.
poly(.beta.-hydroxyalkanoate))), poly(orthoesters),
polycyanoacrylates, polyvinyl alcohols, polyurethanes,
polyphosphazenes, polyacrylates, polymethacrylates, polyureas,
polystyrenes, and polyamines, polylysine, polylysine-PEG
copolymers, and poly(ethyleneimine), poly(ethylene imine)-PEG
copolymers.
[0148] In some embodiments, polymers in accordance with the present
invention include polymers which have been approved for use in
humans by the U.S. Food and Drug Administration (FDA) under 21
C.F.R. .sctn.177.2600, including but not limited to polyesters
(e.g., polylactic acid, poly(lactic-co-glycolic acid),
polycaprolactone, polyvalerolactone, poly(1,3-dioxan-2one));
polyanhydrides (e.g., poly(sebacic anhydride)); polyethers (e.g.,
polyethylene glycol); polyurethanes; polymethacrylates;
polyacrylates; and polycyanoacrylates.
[0149] In some embodiments, polymers can be hydrophilic. For
example, polymers may comprise anionic groups (e.g., phosphate
group, sulphate group, carboxylate group); cationic groups (e.g.,
quaternary amine group); or polar groups (e.g., hydroxyl group,
thiol group, amine group). In some embodiments, a synthetic
nanocarrier comprising a hydrophilic polymeric matrix generates a
hydrophilic environment within the synthetic nanocarrier. In some
embodiments, polymers can be hydrophobic. In some embodiments, a
synthetic nanocarrier comprising a hydrophobic polymeric matrix
generates a hydrophobic environment within the synthetic
nanocarrier. Selection of the hydrophilicity or hydrophobicity of
the polymer may have an impact on the nature of materials that are
incorporated (e.g. coupled) within the synthetic nanocarrier.
[0150] In some embodiments, polymers may be modified with one or
more moieties and/or functional groups. A variety of moieties or
functional groups can be used in accordance with the present
invention. In some embodiments, polymers may be modified with
polyethylene glycol (PEG), with a carbohydrate, and/or with acyclic
polyacetals derived from polysaccharides (Papisov, 2001, ACS
Symposium Series, 786:301). Certain embodiments may be made using
the general teachings of U.S. Pat. No. 5,543,158 to Gref et al., or
WO publication WO2009/051837 by Von Andrian et al.
[0151] In some embodiments, polymers may be modified with a lipid
or fatty acid group. In some embodiments, a fatty acid group may be
one or more of butyric, caproic, caprylic, capric, lauric,
myristic, palmitic, stearic, arachidic, behenic, or lignoceric
acid. In some embodiments, a fatty acid group may be one or more of
palmitoleic, oleic, vaccenic, linoleic, alpha-linoleic,
gamma-linoleic, arachidonic, gadoleic, arachidonic,
eicosapentaenoic, docosahexaenoic, or erucic acid.
[0152] In some embodiments, polymers may be polyesters, including
copolymers comprising lactic acid and glycolic acid units, such as
poly(lactic acid-co-glycolic acid) and poly(lactide-co-glycolide),
collectively referred to herein as "PLGA"; and homopolymers
comprising glycolic acid units, referred to herein as "PGA," and
lactic acid units, such as poly-L-lactic acid, poly-D-lactic acid,
poly-D,L-lactic acid, poly-L-lactide, poly-D-lactide, and
poly-D,L-lactide, collectively referred to herein as "PLA." In some
embodiments, exemplary polyesters include, for example,
polyhydroxyacids; PEG copolymers and copolymers of lactide and
glycolide (e.g., PLA-PEG copolymers, PGA-PEG copolymers, PLGA-PEG
copolymers, and derivatives thereof. In some embodiments,
polyesters include, for example, poly(caprolactone),
poly(caprolactone)-PEG copolymers, poly(L-lactide-co-L-lysine),
poly(serine ester), poly(4-hydroxy-L-proline ester),
poly[.alpha.-(4-aminobutyl)-L-glycolic acid], and derivatives
thereof.
[0153] In some embodiments, a polymer may be PLGA. PLGA is a
biocompatible and biodegradable co-polymer of lactic acid and
glycolic acid, and various forms of PLGA are characterized by the
ratio of lactic acid:glycolic acid. Lactic acid can be L-lactic
acid, D-lactic acid, or D,L-lactic acid. The degradation rate of
PLGA can be adjusted by altering the lactic acid:glycolic acid
ratio. In some embodiments, PLGA to be used in accordance with the
present invention is characterized by a lactic acid:glycolic acid
ratio of approximately 85:15, approximately 75:25, approximately
60:40, approximately 50:50, approximately 40:60, approximately
25:75, or approximately 15:85.
[0154] In some embodiments, polymers may be one or more acrylic
polymers. In certain embodiments, acrylic polymers include, for
example, acrylic acid and methacrylic acid copolymers, methyl
methacrylate copolymers, ethoxyethyl methacrylates, cyanoethyl
methacrylate, aminoalkyl methacrylate copolymer, poly(acrylic
acid), poly(methacrylic acid), methacrylic acid alkylamide
copolymer, poly(methyl methacrylate), poly(methacrylic acid
anhydride), methyl methacrylate, polymethacrylate, poly(methyl
methacrylate) copolymer, polyacrylamide, aminoalkyl methacrylate
copolymer, glycidyl methacrylate copolymers, polycyanoacrylates,
and combinations comprising one or more of the foregoing polymers.
The acrylic polymer may comprise fully-polymerized copolymers of
acrylic and methacrylic acid esters with a low content of
quaternary ammonium groups.
[0155] In some embodiments, polymers can be cationic polymers. In
general, cationic polymers are able to condense and/or protect
negatively charged strands of nucleic acids (e.g. DNA, or
derivatives thereof). Amine-containing polymers such as
poly(lysine) (Zauner et al., 1998, Adv. Drug Del. Rev., 30:97; and
Kabanov et al., 1995, Bioconjugate Chem., 6:7), poly(ethylene
imine) (PEI; Boussif et al., 1995, Proc. Natl. Acad. Sci., USA,
1995, 92:7297), and poly(amidoamine) dendrimers (Kukowska-Latallo
et al., 1996, Proc. Natl. Acad. Sci., USA, 93:4897; Tang et al.,
1996, Bioconjugate Chem., 7:703; and Haensler et al., 1993,
Bioconjugate Chem., 4:372) are positively-charged at physiological
pH, form ion pairs with nucleic acids, and mediate transfection in
a variety of cell lines. In embodiments, the inventive synthetic
nanocarriers may not comprise (or may exclude) cationic
polymers.
[0156] In some embodiments, polymers can be degradable polyesters
bearing cationic side chains (Putnam et al., 1999, Macromolecules,
32:3658; Barrera et al., 1993, J. Am. Chem. Soc., 115:11010; Kwon
et al., 1989, Macromolecules, 22:3250; Lim et al., 1999, J. Am.
Chem. Soc., 121:5633; and Zhou et al., 1990, Macromolecules,
23:3399). Examples of these polyesters include
poly(L-lactide-co-L-lysine) (Barrera et al., 1993, J. Am. Chem.
Soc., 115:11010), poly(serine ester) (Zhou et al., 1990,
Macromolecules, 23:3399), poly(4-hydroxy-L-proline ester) (Putnam
et al., 1999, Macromolecules, 32:3658; and Lim et al., 1999, J. Am.
Chem. Soc., 121:5633), and poly(4-hydroxy-L-proline ester) (Putnam
et al., 1999, Macromolecules, 32:3658; and Lim et al., 1999, J. Am.
Chem. Soc., 121:5633).
[0157] The properties of these and other polymers and methods for
preparing them are well known in the art (see, for example, U.S.
Pat. Nos. 6,123,727; 5,804,178; 5,770,417; 5,736,372; 5,716,404;
6,095,148; 5,837,752; 5,902,599; 5,696,175; 5,514,378; 5,512,600;
5,399,665; 5,019,379; 5,010,167; 4,806,621; 4,638,045; and
4,946,929; Wang et al., 2001, J. Am. Chem. Soc., 123:9480; Lim et
al., 2001, J. Am. Chem. Soc., 123:2460; Langer, 2000, Acc. Chem.
Res., 33:94; Langer, 1999, J. Control. Release, 62:7; and Uhrich et
al., 1999, Chem. Rev., 99:3181). More generally, a variety of
methods for synthesizing certain suitable polymers are described in
Concise Encyclopedia of Polymer Science and Polymeric Amines and
Ammonium Salts, Ed. by Goethals, Pergamon Press, 1980; Principles
of Polymerization by Odian, John Wiley & Sons, Fourth Edition,
2004; Contemporary Polymer Chemistry by Allcock et al.,
Prentice-Hall, 1981; Deming et al., 1997, Nature, 390:386; and in
U.S. Pat. Nos. 6,506,577, 6,632,922, 6,686,446, and 6,818,732.
[0158] In some embodiments, polymers can be linear or branched
polymers. In some embodiments, polymers can be dendrimers. In some
embodiments, polymers can be substantially cross-linked to one
another. In some embodiments, polymers can be substantially free of
cross-links. In some embodiments, polymers can be used in
accordance with the present invention without undergoing a
cross-linking step. It is further to be understood that inventive
synthetic nanocarriers may comprise block copolymers, graft
copolymers, blends, mixtures, and/or adducts of any of the
foregoing and other polymers. Those skilled in the art will
recognize that the polymers listed herein represent an exemplary,
not comprehensive, list of polymers that can be of use in
accordance with the present invention.
[0159] Compositions according to the invention comprise synthetic
nanocarriers in combination with pharmaceutically acceptable
excipients, such as preservatives, buffers, saline, or phosphate
buffered saline. The compositions may be made using conventional
pharmaceutical manufacturing and compounding techniques to arrive
at useful dosage forms. In an embodiment, inventive synthetic
nanocarriers are suspended in sterile saline solution for injection
together with a preservative.
[0160] In embodiments, when preparing synthetic nanocarriers as
carriers, methods for coupling components to the synthetic
nanocarriers may be useful. If the component is a small molecule it
may be of advantage to attach the component to a polymer prior to
the assembly of the synthetic nanocarriers. In embodiments, it may
also be an advantage to prepare the synthetic nanocarriers with
surface groups that are used to couple the component to the
synthetic nanocarrier through the use of these surface groups
rather than attaching the component to a polymer and then using
this polymer conjugate in the construction of synthetic
nanocarriers.
[0161] In certain embodiments, the coupling can be a covalent
linker. In embodiments, peptides according to the invention can be
covalently coupled to the external surface via a 1,2,3-triazole
linker formed by the 1,3-dipolar cycloaddition reaction of azido
groups on the surface of the nanocarrier with antigen or
immunosuppressant containing an alkyne group or by the 1,3-dipolar
cycloaddition reaction of alkynes on the surface of the nanocarrier
with components containing an azido group. Such cycloaddition
reactions are preferably performed in the presence of a Cu(I)
catalyst along with a suitable Cu(I)-ligand and a reducing agent to
reduce Cu(II) compound to catalytic active Cu(I) compound. This
Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) can also be
referred as the click reaction.
[0162] Additionally, the covalent coupling may comprise a covalent
linker that comprises an amide linker, a disulfide linker, a
thioether linker, a hydrazone linker, a hydrazide linker, an imine
or oxime linker, an urea or thiourea linker, an amidine linker, an
amine linker, and a sulfonamide linker.
[0163] An amide linker is formed via an amide bond between an amine
on one component with the carboxylic acid group of a second
component such as the nanocarrier. The amide bond in the linker can
be made using any of the conventional amide bond forming reactions
with suitably protected amino acids or components and activated
carboxylic acid such N-hydroxysuccinimide-activated ester.
[0164] A disulfide linker is made via the formation of a disulfide
(S--S) bond between two sulfur atoms of the form, for instance, of
R1-S--S--R2. A disulfide bond can be formed by thiol exchange of a
component containing thiol/mercaptan group (--SH) with another
activated thiol group on a polymer or nanocarrier or a nanocarrier
containing thiol/mercaptan groups with a component containing
activated thiol group.
[0165] A triazole linker, specifically a 1,2,3-triazole of the
form
##STR00001##
wherein R1 and R2 may be any chemical entities, is made by the
1,3-dipolar cycloaddition reaction of an azide attached to a first
component such as the nanocarrier with a terminal alkyne attached
to a second component such as the immunosuppressant or antigen. The
1,3-dipolar cycloaddition reaction is performed with or without a
catalyst, preferably with Cu(I)-catalyst, which links the two
components through a 1,2,3-triazole function. This chemistry is
described in detail by Sharpless et al., Angew. Chem. Int. Ed.
41(14), 2596, (2002) and Meldal, et al, Chem. Rev., 2008, 108(8),
2952-3015 and is often referred to as a "click" reaction or
CuAAC.
[0166] In embodiments, a polymer containing an azide or alkyne
group, terminal to the polymer chain is prepared. This polymer is
then used to prepare a synthetic nanocarrier in such a manner that
a plurality of the alkyne or azide groups are positioned on the
surface of that nanocarrier. Alternatively, the synthetic
nanocarrier can be prepared by another route, and subsequently
functionalized with alkyne or azide groups. The component is
prepared with the presence of either an alkyne (if the polymer
contains an azide) or an azide (if the polymer contains an alkyne)
group. The component is then allowed to react with the nanocarrier
via the 1,3-dipolar cycloaddition reaction with or without a
catalyst which covalently couples the component to the particle
through the 1,4-disubstituted 1,2,3-triazole linker.
[0167] A thioether linker is made by the formation of a
sulfur-carbon (thioether) bond in the form, for instance, of
R1-S--R2. Thioether can be made by either alkylation of a
thiol/mercaptan (--SH) group on one component such as the component
with an alkylating group such as halide or epoxide on a second
component such as the nanocarrier. Thioether linkers can also be
formed by Michael addition of a thiol/mercaptan group on one
component to an electron-deficient alkene group on a second
component such as a polymer containing a maleimide group or vinyl
sulfone group as the Michael acceptor. In another way, thioether
linkers can be prepared by the radical thiol-ene reaction of a
thiol/mercaptan group on one component with an alkene group on a
second component such as a polymer or nanocarrier.
[0168] A hydrazone linker is made by the reaction of a hydrazide
group on one component with an aldehyde/ketone group on the second
component such as the nanocarrier.
[0169] A hydrazide linker is formed by the reaction of a hydrazine
group on one component with a carboxylic acid group on the second
component such as the nanocarrier. Such reaction is generally
performed using chemistry similar to the formation of amide bond
where the carboxylic acid is activated with an activating
reagent.
[0170] An imine or oxime linker is formed by the reaction of an
amine or N-alkoxyamine (or aminooxy) group on one component with an
aldehyde or ketone group on the second component such as the
nanocarrier.
[0171] An urea or thiourea linker is prepared by the reaction of an
amine group on one component with an isocyanate or thioisocyanate
group on the second component such as the nanocarrier.
[0172] An amidine linker is prepared by the reaction of an amine
group on one component with an imidoester group on the second
component such as the nanocarrier.
[0173] An amine linker is made by the alkylation reaction of an
amine group on one component with an alkylating group such as
halide, epoxide, or sulfonate ester group on the second component
such as the nanocarrier. Alternatively, an amine linker can also be
made by reductive amination of an amine group on one component with
an aldehyde or ketone group on the second component such as the
nanocarrier with a suitable reducing reagent such as sodium
cyanoborohydride or sodium triacetoxyborohydride.
[0174] A sulfonamide linker is made by the reaction of an amine
group on one component with a sulfonyl halide (such as sulfonyl
chloride) group on the second component such as the
nanocarrier.
[0175] A sulfone linker is made by Michael addition of a
nucleophile to a vinyl sulfone. Either the vinyl sulfone or the
nucleophile may be on the surface of the nanocarrier or attached to
a component.
[0176] The component can also be conjugated to the nanocarrier via
non-covalent conjugation methods. For example, a negative charged
antigen or immunosuppressant can be conjugated to a positive
charged nanocarrier through electrostatic adsorption. A component
containing a metal ligand can also be conjugated to a nanocarrier
containing a metal complex via a metal-ligand complex.
[0177] In embodiments, the component can be attached to a polymer,
for example polylactic acid-block-polyethylene glycol, prior to the
assembly of the synthetic nanocarrier or the synthetic nanocarrier
can be formed with reactive or activatable groups on its surface.
In the latter case, the component may be prepared with a group
which is compatible with the attachment chemistry that is presented
by the synthetic nanocarriers' surface. In other embodiments, a
peptide component can be attached to VLPs or liposomes using a
suitable linker. A linker is a compound or reagent that capable of
coupling two molecules together. In an embodiment, the linker can
be a homobifuntional or heterobifunctional reagent as described in
Hermanson 2008. For example, an VLP or liposome synthetic
nanocarrier containing a carboxylic group on the surface can be
treated with a homobifunctional linker, adipic dihydrazide (ADH),
in the presence of EDC to form the corresponding synthetic
nanocarrier with the ADH linker. The resulting ADH linked synthetic
nanocarrier is then conjugated with a peptide component containing
an acid group via the other end of the ADH linker on NC to produce
the corresponding VLP or liposome peptide conjugate.
[0178] For detailed descriptions of available conjugation methods,
see Hermanson G T "Bioconjugate Techniques", 2nd Edition Published
by Academic Press, Inc., 2008. In addition to covalent attachment
the component can be coupled by adsorption to a pre-formed
synthetic nanocarrier or it can be coupled by encapsulation during
the formation of the synthetic nanocarrier.
[0179] Any immunosuppressant as provided herein can be coupled to
the synthetic nanocarrier. Immunosuppressants include, but are not
limited to, statins; mTOR inhibitors, such as rapamycin or a
rapamycin analog; TGF-.beta. signaling agents; TGF-.beta. receptor
agonists; histone deacetylase (HDAC) inhibitors; corticosteroids;
inhibitors of mitochondrial function, such as rotenone; P38
inhibitors; NF-.kappa..beta. inhibitors; adenosine receptor
agonists; prostaglandin E2 agonists; phosphodiesterase inhibitors,
such as phosphodiesterase 4 inhibitor; proteasome inhibitors;
kinase inhibitors; G-protein coupled receptor agonists; G-protein
coupled receptor antagonists; glucocorticoids; retinoids; cytokine
inhibitors; cytokine receptor inhibitors; cytokine receptor
activators; peroxisome proliferator-activated receptor antagonists;
peroxisome proliferator-activated receptor agonists; histone
deacetylase inhibitors; calcineurin inhibitors; phosphatase
inhibitors and oxidized ATPs. Immunosuppressants also include IDO,
vitamin D3, cyclosporine A, aryl hydrocarbon receptor inhibitors,
resveratrol, azathiopurine, 6-mercaptopurine, aspirin, niflumic
acid, estriol, tripolide, interleukins (e.g., IL-1, IL-10),
cyclosporine A, siRNAs targeting cytokines or cytokine receptors
and the like.
[0180] Examples of statins include atorvastatin (LIPITOR.RTM.,
TORVAST.RTM.), cerivastatin, fluvastatin (LESCOL.RTM., LESCOL.RTM.
XL), lovastatin (MEVACOR.RTM., ALTOCOR.RTM., ALTOPREV.RTM.),
mevastatin (COMPACTIN.RTM.), pitavastatin (LIVALO.RTM.,
PIAVA.RTM.), rosuvastatin (PRAVACHOL.RTM., SELEKTINE.RTM.,
LIPOSTAT.RTM.), rosuvastatin (CRESTOR.RTM.), and simvastatin
(ZOCOR.RTM., LIPEX.RTM.).
[0181] Examples of mTOR inhibitors include rapamycin and analogs
thereof (e.g., CCL-779, RAD001, AP23573, C20-methallylrapamycin
(C20-Marap), C16-(S)-butylsulfonamidorapamycin (C16-BSrap),
C16-(S)-3-methylindolerapamycin (C16-iRap) (Bayle et al. Chemistry
& Biology 2006, 13:99-107)), AZD8055, BEZ235 (NVP-BEZ235),
chrysophanic acid (chrysophanol), deforolimus (MK-8669), everolimus
(RAD0001), KU-0063794, PI-103, PP242, temsirolimus, and WYE-354
(available from Selleck, Houston, Tex., USA).
[0182] Examples of TGF-.beta. signaling agents include TGF-.beta.
ligands (e.g., activin A, GDF1, GDF11, bone morphogenic proteins,
nodal, TGF-.beta.s) and their receptors (e.g., ACVR1B, ACVR1C,
ACVR2A, ACVR2B, BMPR2, BMPR1A, BMPR1B, TGF.beta.RI, TGF.beta.RII),
R-SMADS/co-SMADS (e.g., SMAD1, SMAD2, SMAD3, SMAD4, SMAD5, SMAD8),
and ligand inhibitors (e.g., follistatin, noggin, chordin, DAN,
lefty, LTBP1, THBS1, Decorin).
[0183] Examples of inhibitors of mitochondrial function include
atractyloside (dipotassium salt), bongkrekic acid (triammonium
salt), carbonyl cyanide m-chlorophenylhydrazone,
carboxyatractyloside (e.g., from Atractylis gummifera), CGP-37157,
(-)-Deguelin (e.g., from Mundulea sericea), F16, hexokinase II VDAC
binding domain peptide, oligomycin, rotenone, Ru360, SFK1, and
valinomycin (e.g., from Streptomyces fulvissimus) (EMD4Biosciences,
USA).
[0184] Examples of P38 inhibitors include SB-203580
(4-(4-Fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole)-
, SB-239063
(trans-1-(4hydroxycyclohexyl)-4-(fluorophenyl)-5-(2-methoxy-pyrimidin-4-y-
l) imidazole), SB-220025
(5-(2amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinyl)imidazole))-
, and ARRY-797.
[0185] Examples of NF (e.g., NK-.kappa..beta.) inhibitors include
IFRD1, 2-(1,8-naphthyridin-2-yl)-Phenol, 5-aminosalicylic acid, BAY
11-7082, BAY 11-7085, CAPE (Caffeic Acid Phenethylester),
diethylmaleate, IKK-2 Inhibitor IV, IMD 0354, lactacystin, MG-132
[Z-Leu-Leu-Leu-CHO], NF.kappa.B Activation Inhibitor III,
NF-.kappa.B Activation Inhibitor II, JSH-23, parthenolide,
Phenylarsine Oxide (PAO), PPM-18, pyrrolidinedithiocarbamic acid
ammonium salt, QNZ, RO 106-9920, rocaglamide, rocaglamide AL,
rocaglamide C, rocaglamide I, rocaglamide J, rocaglaol, (R)-MG-132,
sodium salicylate, triptolide (PG490), wedelolactone.
[0186] Examples of adenosine receptor agonists include CGS-21680
and ATL-146e.
[0187] Examples of prostaglandin E2 agonists include E-Prostanoid 2
and E-Prostanoid 4.
[0188] Examples of phosphodiesterase inhibitors (non-selective and
selective inhibitors) include caffeine, aminophylline, IBMX
(3-isobutyl-1-methylxanthine), paraxanthine, pentoxifylline,
theobromine, theophylline, methylated xanthines, vinpocetine, EHNA
(erythro-9-(2-hydroxy-3-nonyl)adenine), anagrelide, enoximone
(PERFAN), milrinone, levosimendon, mesembrine, ibudilast,
piclamilast, luteolin, drotaverine, roflumilast (DAXAS.TM.,
DALIRESP.TM.), sildenafil (REVATION.RTM., VIAGRA.RTM.), tadalafil
(ADCIRCA.RTM., CIALIS.RTM.), vardenafil (LEVITRA.RTM.,
STAXYN.RTM.), udenafil, avanafil, icariin, 4-methylpiperazine, and
pyrazolo pyrimidin-7-1.
[0189] Examples of proteasome inhibitors include bortezomib,
disulfiram, epigallocatechin-3-gallate, and salinosporamide A.
[0190] Examples of kinase inhibitors include bevacizumab, BIBW
2992, cetuximab (ERBITUX.RTM.), imatinib (GLEEVEC.RTM.),
trastuzumab (HERCEPTIN.RTM.), gefitinib (IRESSA.RTM.), ranibizumab
(LUCENTIS.RTM.), pegaptanib, sorafenib, dasatinib, sunitinib,
erlotinib, nilotinib, lapatinib, panitumumab, vandetanib, E7080,
pazopanib, mubritinib.
[0191] Examples of glucocorticoids include hydrocortisone
(cortisol), cortisone acetate, prednisone, prednisolone,
methylprednisolone, dexamethasone, betamethasone, triamcinolone,
beclometasone, fludrocortisone acetate, deoxycorticosterone acetate
(DOCA), and aldosterone.
[0192] Examples of retinoids include retinol, retinal, tretinoin
(retinoic acid, RETIN-A.RTM.), isotretinoin (ACCUTANE.RTM.,
AMNESTEEM.RTM., CLARAVIS.RTM., SOTRET.RTM.), alitretinoin
(PANRETIN.RTM.), etretinate (TEGISON) and its metabolite acitretin
(SORIATANE.RTM.), tazarotene (TAZORAC.RTM., AVAGE.RTM.,
ZORAC.RTM.), bexarotene (TARGRETIN.RTM.), and adapalene
(DIFFERIN.RTM.).
[0193] Examples of cytokine inhibitors include IL1ra, IL1 receptor
antagonist, IGFBP, TNF-BF, uromodulin, Alpha-2-Macroglobulin,
Cyclosporin A, Pentamidine, and Pentoxifylline (PENTOPAK.RTM.,
PENTOXIL.RTM., TRENTAL.RTM.).
[0194] Examples of peroxisome proliferator-activated receptor
antagonists include GW9662, PPAR.gamma. antagonist III, G335,
T0070907 (EMD4Biosciences, USA).
[0195] Examples of peroxisome proliferator-activated receptor
agonists include pioglitazone, ciglitazone, clofibrate, GW1929,
GW7647, L-165,041, LY 171883, PPAR.gamma. activator, Fmoc-Leu,
troglitazone, and WY-14643 (EMD4Biosciences, USA).
[0196] Examples of histone deacetylase inhibitors include
hydroxamic acids (or hydroxamates) such as trichostatin A, cyclic
tetrapeptides (such as trapoxin B) and depsipeptides, benzamides,
electrophilic ketones, aliphatic acid compounds such as
phenylbutyrate and valproic acid, hydroxamic acids such as
vorinostat (SAHA), belinostat (PXD101), LAQ824, and panobinostat
(LBH589), benzamides such as entinostat (MS-275), CI994, and
mocetinostat (MGCD0103), nicotinamide, derivatives of NAD,
dihydrocoumarin, naphthopyranone, and 2-hydroxynaphaldehydes.
[0197] Examples of calcineurin inhibitors include cyclosporine,
pimecrolimus, voclosporin, and tacrolimus.
[0198] Examples of phosphatase inhibitors include BN82002
hydrochloride, CP-91149, calyculin A, cantharidic acid,
cantharidin, cypermethrin, ethyl-3,4-dephostatin, fostriecin sodium
salt, MAZ51, methyl-3,4-dephostatin, NSC 95397, norcantharidin,
okadaic acid ammonium salt from prorocentrum concavum, okadaic
acid, okadaic acid potassium salt, okadaic acid sodium salt,
phenylarsine oxide, various phosphatase inhibitor cocktails,
protein phosphatase 1C, protein phosphatase 2A inhibitor protein,
protein phosphatase 2A1, protein phosphatase 2A2, sodium
orthovanadate.
[0199] In some embodiments, antigens as described herein are also
coupled to synthetic nanocarriers. In some embodiments, the
antigens are coupled to the same or different synthetic
nanocarriers as to which the immunosuppressants are coupled. In
other embodiments, the antigens are not coupled to any synthetic
nanocarriers. Antigens include any of the antigens provided herein,
or fragments or derivatives thereof, such antigens are associated
with inflammatory, autoimmune diseases, allergy, organ or tissue
rejection, graft versus host disease, transplant antigens and
therapeutic protein antigens. The epitopes, or proteins,
polypeptides or peptides that comprise the epitopes, can be
obtained or derived from any of the antigens provided or otherwise
known in the art.
[0200] Therapeutic proteins include, but are not limited to,
infusible therapeutic proteins, enzymes, enzyme cofactors,
hormones, blood clotting factors, cytokines and interferons, growth
factors, monoclonal antibodies, and polyclonal antibodies (e.g.,
that are administered to a subject as a replacement therapy), and
proteins associated with Pompe's disease (e.g., alglucosidase alfa,
rhGAA (e.g., Myozyme and Lumizyme (Genzyme)). Therapeutic proteins
also include proteins involved in the blood coagulation cascade.
Therapeutic proteins include, but are not limited to, Factor VIII,
Factor VII, Factor IX, Factor V, von Willebrand Factor, von
Heldebrant Factor, tissue plasminogen activator, insulin, growth
hormone, erythropoietin alfa, VEGF, thrombopoietin, lysozyme,
antithrombin and the like. Therapeutic proteins also include
adipokines, such as leptin and adiponectin. Other examples of
therapeutic proteins are as described below and elsewhere herein.
Also included are fragments or derivatives of any of the
therapeutic proteins provided as the antigen.
[0201] Examples of therapeutic proteins used in enzyme replacement
therapy of subjects having a lysosomal storage disorder include,
but are not limited to, imiglucerase for the treatment of Gaucher's
disease (e.g., CEREZYME.TM.), a-galactosidase A (a-gal A) for the
treatment of Fabry disease (e.g., agalsidase beta, FABRYZYME.TM.),
acid a-glucosidase (GAA) for the treatment of Pompe disease (e.g.,
alglucosidase alfa, LUMIZYME.TM., MYOZYME.TM.), arylsulfatase B for
the treatment of Mucopolysaccharidoses (e.g., laronidase,
ALDURAZYME.TM., idursulfase, ELAPRASE.TM., arylsulfatase B,
NAGLAZYME.TM.).
[0202] Examples of enzymes include oxidoreductases, transferases,
hydrolases, lyases, isomerases, and ligases.
[0203] Examples of hormones include Melatonin
(N-acetyl-5-methoxytryptamine), Serotonin, Thyroxine (or
tetraiodothyronine) (a thyroid hormone), Triiodothyronine (a
thyroid hormone), Epinephrine (or adrenaline), Norepinephrine (or
noradrenaline), Dopamine (or prolactin inhibiting hormone),
Antimullerian hormone (or mullerian inhibiting factor or hormone),
Adiponectin, Adrenocorticotropic hormone (or corticotropin),
Angiotensinogen and angiotensin, Antidiuretic hormone (or
vasopressin, arginine vasopressin), Atrial-natriuretic peptide (or
atriopeptin), Calcitonin, Cholecystokinin, Corticotropin-releasing
hormone, Erythropoietin, Follicle-stimulating hormone, Gastrin,
Ghrelin, Glucagon, Glucagon-like peptide (GLP-1), GIP,
Gonadotropin-releasing hormone, Growth hormone-releasing hormone,
Human chorionic gonadotropin, Human placental lactogen, Growth
hormone, Inhibin, Insulin, Insulin-like growth factor (or
somatomedin), Leptin, Luteinizing hormone, Melanocyte stimulating
hormone, Orexin, Oxytocin, Parathyroid hormone, Prolactin, Relaxin,
Secretin, Somatostatin, Thrombopoietin, Thyroid-stimulating hormone
(or thyrotropin), Thyrotropin-releasing hormone, Cortisol,
Aldosterone, Testosterone, Dehydroepiandrosterone, Androstenedione,
Dihydrotestosterone, Estradiol, Estrone, Estriol, Progesterone,
Calcitriol (1,25-dihydroxyvitamin D3), Calcidiol (25-hydroxyvitamin
D3), Prostaglandins, Leukotrienes, Prostacyclin, Thromboxane,
Prolactin releasing hormone, Lipotropin, Brain natriuretic peptide,
Neuropeptide Y, Histamine, Endothelin, Pancreatic polypeptide,
Renin, and Enkephalin.
[0204] Examples of blood and blood coagulation factors include
Factor I (fibrinogen), Factor II (prothrombin), tissue factor,
Factor V (proaccelerin, labile factor), Factor VII (stable factor,
proconvertin), Factor VIII (antihemophilic globulin), Factor IX
(Christmas factor or plasma thromboplastin component), Factor X
(Stuart-Prower factor), Factor Xa, Factor XI, Factor XII (Hageman
factor), Factor XIII (fibrin-stabilizing factor), von Willebrand
factor, prekallikrein (Fletcher factor), high-molecular weight
kininogen (HMWK) (Fitzgerald factor), fibronectin, fibrin,
thrombin, antithrombin III, heparin cofactor II, protein C, protein
S, protein Z, protein Z-related protease inhibitot (ZPI),
plasminogen, alpha 2-antiplasmin, tissue plasminogen activator
(tPA), urokinase, plasminogen activator inhibitor-1 (PAI1),
plasminogen activator inhibitor-2 (PAI2), cancer procoagulant, and
epoetin alfa (Epogen, Procrit).
[0205] Examples of cytokines include lymphokines, interleukins, and
chemokines, type 1 cytokines, such as IFN-.gamma., TGF-.beta., and
type 2 cytokines, such as IL-4, IL-10, and IL-13.
[0206] Examples of growth factors include Adrenomedullin (AM),
Angiopoietin (Ang), Autocrine motility factor, Bone morphogenetic
proteins (BMPs), Brain-derived neurotrophic factor (BDNF),
Epidermal growth factor (EGF), Erythropoietin (EPO), Fibroblast
growth factor (FGF), Glial cell line-derived neurotrophic factor
(GDNF), Granulocyte colony-stimulating factor (G-CSF), Granulocyte
macrophage colony-stimulating factor (GM-CSF), Growth
differentiation factor-9 (GDF9), Hepatocyte growth factor (HGF),
Hepatoma-derived growth factor (HDGF), Insulin-like growth factor
(IGF), Migration-stimulating factor, Myostatin (GDF-8), Nerve
growth factor (NGF) and other neurotrophins, Platelet-derived
growth factor (PDGF), Thrombopoietin (TPO), Transforming growth
factor alpha(TGF-.alpha.), Transforming growth factor
beta(TGF-.beta.), Tumour_necrosis_factor-alpha(TNF-.alpha.),
Vascular endothelial growth factor (VEGF), Wnt Signaling Pathway,
placental growth factor (P1GF), [(Foetal Bovine Somatotrophin)]
(FBS), IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, and IL-7.
[0207] Examples of monoclonal antibodies include Abagovomab,
Abciximab, Adalimumab, Adecatumumab, Afelimomab, Afutuzumab,
Alacizumab pegol, ALD, Alemtuzumab, Altumomab pentetate, Anatumomab
mafenatox, Anrukinzumab, Anti-thymocyte globin, Apolizumab,
Arcitumomab, Aselizumab, Atlizumab (tocilizumab), Atorolimumab,
Bapineuzumab, Basiliximab, Bavituximab, Bectumomab, Belimumab,
Benralizumab, Bertilimumab, Besilesomab, Bevacizumab, Biciromab,
Bivatuzumab mertansine, Blinatumomab, Brentuximab vedotin,
Briakinumab, Canakinumab, Cantuzumab mertansine, Capromab
pendetide, Catumaxomab, Cedelizumab, Certolizumab pegol, Cetuximab,
Citatuzumab bogatox, Cixutumumab, Clenoliximab, Clivatuzumab
tetraxetan, Conatumumab, Dacetuzumab, Daclizumab, Daratumumab,
Denosumab, Detumomab, Dorlimomab aritox, Dorlixizumab, Ecromeximab,
Eculizumab, Edobacomab, Edrecolomab, Efalizumab, Efungumab,
Elotuzumab, Elsilimomab, Enlimomab pegol, Epitumomab cituxetan,
Epratuzumab, Erlizumab, Ertumaxomab, Etaracizumab, Exbivirumab,
Fanolesomab, Faralimomab, Farletuzumab, Felvizumab, Fezakinumab,
Figitumumab, Fontolizumab, Foravirumab, Fresolimumab, Galiximab,
Gantenerumab, Gavilimomab, Gemtuzumab ozogamicin, GC1008,
Girentuximab, Glembatumumab vedotin, Golimumab, Gomiliximab,
Ibalizumab, Ibritumomab tiuxetan, Igovomab, Imciromab, Infliximab,
Intetumumab, Inolimomab, Inotuzumab ozogamicin, Ipilimumab,
Iratumumab, Keliximab, Labetuzumab, Lebrikizumab, Lemalesomab,
Lerdelimumab, Lexatumumab, Libivirumab, Lintuzumab, Lorvotuzumab
mertansine, Lucatumumab, Lumiliximab, Mapatumumab, Maslimomab,
Matuzumab, Mepolizumab, Metelimumab, Milatuzumab, Minretumomab,
Mitumomab, Morolimumab, Motavizumab, Muromonab-CD3, Nacolomab
tafenatox, Naptumomab estafenatox, Natalizumab, Nebacumab,
Necitumumab, Nerelimomab, Nimotuzumab, Nofetumomab merpentan,
Ocrelizumab, Odulimomab, Ofatumumab, Olaratumab, Omalizumab,
Oportuzumab monatox, Oregovomab, Otelixizumab, Pagibaximab,
Palivizumab, Panitumumab, Panobacumab, Pascolizumab, Pemtumomab,
Pertuzumab, Pexelizumab, Pintumomab, Priliximab, Pritumumab,
Rafivirumab, Ramucirumab, Ranibizumab, Raxibacumab, Regavirumab
Reslizumab, Rilotumumab, Rituximab, Robatumumab, Rontalizumab,
Rovelizumab, Ruplizumab, Satumomab pendetide, Sevirumab,
Sibrotuzumab, Sifalimumab, Siltuximab, Siplizumab, Solanezumab,
Sonepcizumab, Sontuzumab, Stamulumab, Sulesomab, Tacatuzumab
tetraxetan, Tadocizumab, Talizumab, Tanezumab, Taplitumomab paptox,
Tefibazumab, Telimomab aritox, Tenatumomab, Teneliximab,
Teplizumab, Ticilimumab (tremelimumab), Tigatuzumab, Tocilizumab
(atlizumab), Toralizumab, Tositumomab, Trastuzumab, Tremelimumab,
Tucotuzumab celmoleukin, Tuvirumab, Urtoxazumab, Ustekinumab,
Vapaliximab, Vedolizumab, Veltuzumab, Vepalimomab, Visilizumab,
Volociximab, Votumumab, Zalutumumab, Zanolimumab, Ziralimumab, and
Zolimomab aritox.
[0208] Examples of infusion therapy or injectable therapeutic
proteins include, for example, Tocilizumab (Roche/Actemra.RTM.),
alpha-1 antitryp sin (Kamada/AAT), Hematide.RTM. (Affymax and
Takeda, synthetic peptide), albinterferon alfa-2b
(Novartis/Zalbin.TM.), Rhucin.RTM. (Pharming Group, C1 inhibitor
replacement therapy), tesamorelin (Theratechnologies/Egrifta,
synthetic growth hormone-releasing factor), ocrelizumab (Genentech,
Roche and Biogen), belimumab (GlaxoSmithKline/Benlysta.RTM.),
pegloticase (Savient Pharmaceuticals/Krystexxa.TM.), taliglucerase
alfa (Protalix/Uplyso), agalsidase alfa (Shire/Replagal.RTM.),
velaglucerase alfa (Shire).
[0209] Additional therapeutic proteins useful in accordance to
aspects of this invention will be apparent to those of skill in the
art, and the invention is not limited in this respect.
[0210] In some embodiments, a component, such as an antigen or
immunosuppressant, may be isolated. Isolated refers to the element
being separated from its native environment and present in
sufficient quantities to permit its identification or use. This
means, for example, the element may be (i) selectively produced by
expression cloning or (ii) purified as by chromatography or
electrophoresis. Isolated elements may be, but need not be,
substantially pure. Because an isolated element may be admixed with
a pharmaceutically acceptable excipient in a pharmaceutical
preparation, the element may comprise only a small percentage by
weight of the preparation. The element is nonetheless isolated in
that it has been separated from the substances with which it may be
associated in living systems, i.e., isolated from other lipids or
proteins. Any of the elements provided herein can be included in
the compositions in isolated form.
D. METHODS OF MAKING AND USING THE INVENTIVE COMPOSITIONS AND
RELATED METHODS
[0211] Synthetic nanocarriers may be prepared using a wide variety
of methods known in the art. For example, synthetic nanocarriers
can be formed by methods as nanoprecipitation, flow focusing using
fluidic channels, spray drying, single and double emulsion solvent
evaporation, solvent extraction, phase separation, milling,
microemulsion procedures, microfabrication, nanofabrication,
sacrificial layers, simple and complex coacervation, and other
methods well known to those of ordinary skill in the art.
Alternatively or additionally, aqueous and organic solvent
syntheses for monodisperse semiconductor, conductive, magnetic,
organic, and other nanomaterials have been described (Pellegrino et
al., 2005, Small, 1:48; Murray et al., 2000, Ann. Rev. Mat. Sci.,
30:545; and Trindade et al., 2001, Chem. Mat., 13:3843). Additional
methods have been described in the literature (see, e.g., Doubrow,
Ed., "Microcapsules and Nanoparticles in Medicine and Pharmacy,"
CRC Press, Boca Raton, 1992; Mathiowitz et al., 1987, J. Control.
Release, 5:13; Mathiowitz et al., 1987, Reactive Polymers, 6:275;
and Mathiowitz et al., 1988, J. Appl. Polymer Sci., 35:755; U.S.
Pat. Nos. 5,578,325 and 6,007,845; P. Paolicelli et al.,
"Surface-modified PLGA-based Nanoparticles that can Efficiently
Associate and Deliver Virus-like Particles" Nanomedicine.
5(6):843-853 (2010)).
[0212] Various materials may be encapsulated into synthetic
nanocarriers as desirable using a variety of methods including but
not limited to C. Astete et al., "Synthesis and characterization of
PLGA nanoparticles" J. Biomater. Sci. Polymer Edn, Vol. 17, No. 3,
pp. 247-289 (2006); K. Avgoustakis "Pegylated Poly(Lactide) and
Poly(Lactide-Co-Glycolide) Nanoparticles: Preparation, Properties
and Possible Applications in Drug Delivery" Current Drug Delivery
1:321-333 (2004); C. Reis et al., "Nanoencapsulation I. Methods for
preparation of drug-loaded polymeric nanoparticles" Nanomedicine
2:8-21 (2006); P. Paolicelli et al., "Surface-modified PLGA-based
Nanoparticles that can Efficiently Associate and Deliver Virus-like
Particles" Nanomedicine. 5(6):843-853 (2010). Other methods
suitable for encapsulating materials into synthetic nanocarriers
may be used, including without limitation methods disclosed in U.S.
Pat. No. 6,632,671 to Unger Oct. 14, 2003.
[0213] In certain embodiments, synthetic nanocarriers are prepared
by a nanoprecipitation process or spray drying. Conditions used in
preparing synthetic nanocarriers may be altered to yield particles
of a desired size or property (e.g., hydrophobicity,
hydrophilicity, external morphology, "stickiness," shape, etc.).
The method of preparing the synthetic nanocarriers and the
conditions (e.g., solvent, temperature, concentration, air flow
rate, etc.) used may depend on the materials to be coupled to the
synthetic nanocarriers and/or the composition of the polymer
matrix.
[0214] If particles prepared by any of the above methods have a
size range outside of the desired range, particles can be sized,
for example, using a sieve.
[0215] Elements (i.e., components) of the inventive synthetic
nanocarriers (such as moieties of which an immunofeature surface is
comprised, targeting moieties, polymeric matrices, antigens,
immunosuppressants and the like) may be coupled to the overall
synthetic nanocarrier, e.g., by one or more covalent bonds, or may
be coupled by means of one or more linkers. Additional methods of
functionalizing synthetic nanocarriers may be adapted from
Published US Patent Application 2006/0002852 to Saltzman et al.,
Published US Patent Application 2009/0028910 to DeSimone et al., or
Published International Patent Application WO/2008/127532 A1 to
Murthy et al.
[0216] Alternatively or additionally, synthetic nanocarriers can be
coupled to components directly or indirectly via non-covalent
interactions. In non-covalent embodiments, the non-covalent
coupling is mediated by non-covalent interactions including but not
limited to charge interactions, affinity interactions, metal
coordination, physical adsorption, host-guest interactions,
hydrophobic interactions, TT stacking interactions, hydrogen
bonding interactions, van der Waals interactions, magnetic
interactions, electrostatic interactions, dipole-dipole
interactions, and/or combinations thereof. Such couplings may be
arranged to be on an external surface or an internal surface of an
inventive synthetic nanocarrier. In embodiments, encapsulation
and/or absorption is a form of coupling. In embodiments, the
inventive synthetic nanocarriers can be combined with an antigen by
admixing in the same vehicle or delivery system.
[0217] Populations of synthetic nanocarriers may be combined to
form pharmaceutical dosage forms according to the present invention
using traditional pharmaceutical mixing methods. These include
liquid-liquid mixing in which two or more suspensions, each
containing one or more subsets of nanocarriers, are directly
combined or are brought together via one or more vessels containing
diluent. As synthetic nanocarriers may also be produced or stored
in a powder form, dry powder-powder mixing could be performed as
could the re-suspension of two or more powders in a common media.
Depending on the properties of the nanocarriers and their
interaction potentials, there may be advantages conferred to one or
another route of mixing.
[0218] Typical inventive compositions that comprise synthetic
nanocarriers may comprise inorganic or organic buffers (e.g.,
sodium or potassium salts of phosphate, carbonate, acetate, or
citrate) and pH adjustment agents (e.g., hydrochloric acid, sodium
or potassium hydroxide, salts of citrate or acetate, amino acids
and their salts) antioxidants (e.g., ascorbic acid,
alpha-tocopherol), surfactants (e.g., polysorbate 20, polysorbate
80, polyoxyethylene9-10 nonyl phenol, sodium desoxycholate),
solution and/or cryo/lyo stabilizers (e.g., sucrose, lactose,
mannitol, trehalose), osmotic adjustment agents (e.g., salts or
sugars), antibacterial agents (e.g., benzoic acid, phenol,
gentamicin), antifoaming agents (e.g., polydimethylsilozone),
preservatives (e.g., thimerosal, 2-phenoxyethanol, EDTA), polymeric
stabilizers and viscosity-adjustment agents (e.g.,
polyvinylpyrrolidone, poloxamer 488, carboxymethylcellulose) and
co-solvents (e.g., glycerol, polyethylene glycol, ethanol).
[0219] Compositions according to the invention comprise inventive
synthetic nanocarriers in combination with pharmaceutically
acceptable excipients. The compositions may be made using
conventional pharmaceutical manufacturing and compounding
techniques to arrive at useful dosage forms. Techniques suitable
for use in practicing the present invention may be found in
Handbook of Industrial Mixing: Science and Practice, Edited by
Edward L. Paul, Victor A. Atiemo-Obeng, and Suzanne M. Kresta, 2004
John Wiley & Sons, Inc.; and Pharmaceutics: The Science of
Dosage Form Design, 2nd Ed. Edited by M. E. Auten, 2001, Churchill
Livingstone. In an embodiment, inventive synthetic nanocarriers are
suspended in sterile saline solution for injection together with a
preservative.
[0220] It is to be understood that the compositions of the
invention can be made in any suitable manner, and the invention is
in no way limited to compositions that can be produced using the
methods described herein. Selection of an appropriate method may
require attention to the properties of the particular moieties
being associated.
[0221] In some embodiments, inventive synthetic nanocarriers are
manufactured under sterile conditions or are terminally sterilized.
This can ensure that resulting compositions are sterile and
non-infectious, thus improving safety when compared to non-sterile
compositions. This provides a valuable safety measure, especially
when subjects receiving synthetic nanocarriers have immune defects,
are suffering from infection, and/or are susceptible to infection.
In some embodiments, inventive synthetic nanocarriers may be
lyophilized and stored in suspension or as lyophilized powder
depending on the formulation strategy for extended periods without
losing activity.
[0222] The compositions of the invention can be administered by a
variety of routes, including but not limited to subcutaneous,
intranasal, oral, intravenous, intraperitoneal, intramuscular,
transmucosal, transmuco sal, sublingual, rectal, ophthalmic,
pulmonary, intradermal, transdermal, transcutaneous or intradermal
or by a combination of these routes. Routes of administration also
include administration by inhalation or pulmonary aerosol.
Techniques for preparing aerosol delivery systems are well known to
those of skill in the art (see, for example, Sciarra and Cutie,
"Aerosols," in Remington's Pharmaceutical Sciences, 18th edition,
1990, pp. 1694-1712; incorporated by reference).
[0223] The transplantable grafts or therapeutic proteins provided
as a cell-based therapy of the invention may be administered by
parenteral, intraarterial, intranasal or intravenous administration
or by injection to lymph nodes or anterior chamber of the eye or by
local administration to an organ or tissue of interest. The
administration may be by subcutaneous, intrathecal,
intraventricular, intramuscular, intraperitoneal, intracoronary,
intrapancreatic, intrahepatic or bronchial injection.
[0224] The compositions of the invention can be administered in
effective amounts, such as the effective amounts described
elsewhere herein. Doses of dosage forms contain varying amounts of
populations of synthetic nanocarriers and/or varying amounts of
immunosuppressants and/or antigens, according to the invention. The
amount of synthetic nanocarriers and/or immunosuppressants and/or
antigens present in the inventive dosage forms can be varied
according to the nature of the antigens, the therapeutic benefit to
be accomplished, and other such parameters. In embodiments, dose
ranging studies can be conducted to establish optimal therapeutic
amount of the population of synthetic nanocarriers and the amount
of immunosuppressants and/or antigens to be present in the dosage
form. In embodiments, the synthetic nanocarriers and/or the
immunosuppressants and/or antigens are present in the dosage form
in an amount effective to generate a tolerogenic immune response to
the antigens upon administration to a subject. It may be possible
to determine amounts of the immunosuppressants and/or antigens
effective to generate a tolerogenic immune response using
conventional dose ranging studies and techniques in subjects.
Inventive dosage forms may be administered at a variety of
frequencies. In a preferred embodiment, at least one administration
of the dosage form is sufficient to generate a pharmacologically
relevant response. In more preferred embodiments, at least two
administrations, at least three administrations, or at least four
administrations, of the dosage form are utilized to ensure a
pharmacologically relevant response.
[0225] Prophylactic administration of the inventive compositions
can be initiated prior to the onset of disease, disorder or
condition or therapeutic administration can be initiated after a
disorder, disorder or condition is established.
[0226] In some embodiments, administration of synthetic
nanocarriers is undertaken e.g., prior to administration of a
therapeutic protein or transplantable graft or exposure to an
allergen. In exemplary embodiments, synthetic nanocarriers are
administered at one or more times including, but not limited to,
30, 25, 20, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0
days prior to administration of a therapeutic protein or
transplantable graft or exposure to an allergen. In addition or
alternatively, synthetic nanocarriers can be administered to a
subject following administration of a therapeutic protein or
transplantable graft or exposure to an allergen. In exemplary
embodiments, synthetic nanocarriers are administered at one or more
times including, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 20, 25, 30, etc. days following administration
of a therapeutic protein or transplantable graft or exposure to an
allergen.
[0227] In some embodiments, a maintenance dose (e.g., of a
synthetic nanocarrier composition provided herein) is administered
to a subject after an initial administration has resulted in a
tolerogenic response in the subject, for example to maintain the
tolerogenic effect achieved after the initial dose, to prevent an
undesired immune reaction in the subject, or to prevent the subject
becoming a subject at risk of experiencing an undesired immune
response or an undesired level of an immune response. In some
embodiments, the maintenance dose is the same dose as the initial
dose the subject received. In some embodiments, the maintenance
dose is a lower dose than the initial dose. For example, in some
embodiments, the maintenance dose is about 3/4, about 2/3, about
1/2, about 1/3, about 1/4, about 1/8, about 1/10, about 1/20, about
1/25, about 1/50, about 1/100, about 1/1,000, about 1/10,000, about
1/100,000, or about 1/1,000,000 (weight/weight) of the initial
dose.
[0228] The compositions and methods described herein can be used to
induce or enhance a tolerogenic immune response and/or to suppress,
modulate, direct or redirect an undesired immune response for the
purpose of immune suppression. The compositions and methods
described herein can be used in the diagnosis, prophylaxis and/or
treatment of diseases, disorders or conditions in which immune
suppression (e.g., tolerogenic immune response) would confer a
treatment benefit. Such diseases, disorders or conditions include
inflammatory diseases, autoimmune diseases, allergies, organ or
tissue rejection and graft versus host disease. The compositions
and methods described herein can also be used in subjects who have
undergone or will undergo transplantation. The compositions and
methods described herein can also be used in subjects who have
received, are receiving or will receive a therapeutic protein
against which they have generated or are expected to generate an
undesired immune response.
[0229] Autoimmune diseases include, but are not limited to,
rheumatoid arthritis, multiple sclerosis, immune-mediated or Type I
diabetes mellitus, inflammatory bowel disease (e.g., Crohn's
disease or ulcerative colitis), systemic lupus erythematosus,
psoriasis, scleroderma, autoimmune thyroid disease, alopecia
areata, Grave's disease, Guillain-Barre syndrome, celiac disease,
Sjogren's syndrome, rheumatic fever, gastritis, autoimmune atrophic
gastritis, autoimmune hepatitis, insulitis, oophoritis, orchitis,
uveitis, phacogenic uveitis, myasthenia gravis, primary myxoedema,
pernicious anemia, autoimmune haemolytic anemia, Addison's disease,
scleroderma, Goodpasture's syndrome, nephritis, for example,
glomerulonephritis, psoriasis, pemphigus vulgaris, pemphigoid,
sympathetic opthalmia, idiopathic thrombocylopenic purpura,
idiopathic feucopenia, Wegener's granulomatosis and
poly/dermatomyositis.
[0230] Some additional exemplary autoimmune diseases, associated
autoantigens, and autoantibodies, which are contemplated for use in
the invention, are described in Table 1 below:
TABLE-US-00001 Autoantibody Type Autoantibody Autoantigen
Autoimmune disease or disorder Antinuclear Anti-SSA/Ro
ribonucleoproteins Systemic lupus erythematosus, neonatal
antibodies autoantibodies heart block, primary Sjogren's syndrome
Anti-La/SS-B ribonucleoproteins Primary Sjogren's syndrome
autoantibodies Anti-centromere centromere CREST syndrome antibodies
Anti-neuronal Ri[disambiguation Opsoclonus nuclear antibody-2
needed] Anti-dsDNA double-stranded SLE DNA Anti-Jol histidine-tRNA
Inflammatory myopathy ligase Anti-Smith snRNP core proteins SLE
Anti- Type I Systemic sclerosis topoisomerase topoisomerase
(anti-Scl-70 antibodies) antibodies Anti-histone histones SLE and
Drug-induced LE[2] antibodies Anti-p62 nucleoporin 62 Primary
biliary cirrhosis[3][4][5] antibodies[3] Anti-sp100 Sp100 nuclear
antibodies [4] antigen Anti-glycoprotein- nucleoporin 210 kDa 210
antibodies[5] Anti-transglutaminase Anti-tTG Coeliac disease
antibodies Anti-eTG Dermatitis herpetiformis Anti-ganglioside
ganglioside GQ1B Miller-Fisher Syndrome antibodies ganglioside GD3
Acute motor axonal neuropathy (AMAN) ganglioside GM1 Multifocal
motor neuropathy with conduction block (MMN) Anti-actin antibodies
actin Coeliac disease anti-actin antibodies correlated with the
level of intestinal damage [6][7] Liver kidney Autoimmune
hepatitis.[8] microsomal type 1 antibody Lupus anticoagulant
Anti-thrombin thrombin Systemic lupus erythematosus antibodies
Anti-neutrophil phospholipid Antiphospholipid syndrome cytoplasmic
antibody c-ANCA proteins in Wegener's granulomatosis neutrophil
cytoplasm p-ANCA neutrophil Microscopic polyangiitis, Churg-Strauss
perinuclear syndrome, systemic vasculitides (non- specific)
Rheumatoid factor IgG Rheumatoid arthritis Anti-smooth muscle
smooth muscle Chronic autoimmune hepatitis antibody
Anti-mitochondrial mitochondria Primary biliary cirrhosis[9]
antibody Anti-SRP signal recognition Polymyositis[10] particle
exosome complex Scleromyositis nicotinic Myasthenia gravis
acetylcholine receptor muscle-specific Myasthenia gravis kinase
(MUSK) Anti-VGCC voltage-gated Lambert-Eaton myasthenic syndrome
calcium channel (P/Q-type) thyroid peroxidase Hashimoto's
thyroiditis (microsomal) TSH receptor Graves' disease Hu
Paraneoplastic cerebellar syndrome Yo (cerebellar Paraneoplastic
cerebellar syndrome Purkinje Cells) amphiphysin Stiff person
syndrome, paraneoplastic cerebellar syndrome Anti-VGKC
voltage-gated Limbic encephalitis, Isaac's Syndrome potassium
channel (autoimmune neuromyotonia) (VGKC) basal ganglia Sydenham's
chorea, paediatric autoimmune neurons neuropsychiatric disease
associated with Streptococcus (PANDAS) N-methyl-D- Encephalitis
aspartate receptor (NMDA) glutamic acid Diabetes mellitus type 1,
stiff person decarboxylase syndrome (GAD) aquaporin-4 Neuromyelitis
optica (Devic's syndrome)
[0231] Inflammatory diseases include, but are not limited to,
Alzheimer's, Ankylosing spondylitis, arthritis, asthma,
atherosclerosis, Behcet's disease, chronic inflammatory
demyelinating polyradiculoneuropathy, Crohn's disease, colitis,
cystic fibrosis, dermatitis, diverticulitis, hepatitis, irritable
bowel syndrome (IBS), lupus erythematous, muscular dystrophy,
nephritis, Parkinson's, shingles and ulcerative colitis.
Inflammatory diseases also include, for example, cardiovascular
disease, chronic obstructive pulmonary disease (COPD),
bronchiectasis, chronic cholecystitis, tuberculosis, Hashimoto's
thyroiditis, sepsis, sarcoidosis, silicosis and other
pneumoconioses, and an implanted foreign body in a wound, but are
not so limited. As used herein, the term "sepsis" refers to a
well-recognized clinical syndrome associated with a host's systemic
inflammatory response to microbial invasion. The term "sepsis" as
used herein refers to a condition that is typically signaled by
fever or hypothermia, tachycardia, and tachypnea, and in severe
instances can progress to hypotension, organ dysfunction, and even
death.
[0232] In some embodiments, the inflammatory disease is
non-autoimmune inflammatory bowel disease, post-surgical adhesions,
coronary artery disease, hepatic fibrosis, acute respiratory
distress syndrome, acute inflammatory pancreatitis, endoscopic
retrograde cholangiopancreatography-induced pancreatitis, burns,
atherogenesis of coronary, cerebral and peripheral arteries,
appendicitis, cholecystitis, diverticulitis, visceral fibrotic
disorders, wound healing, skin scarring disorders (keloids,
hidradenitis suppurativa), granulomatous disorders (sarcoidosis,
primary biliary cirrhosis), asthma, pyoderma gandrenosum, Sweet's
syndrome, Behcet's disease, primary sclerosing cholangitis or an
abscess. In some preferred embodiment the inflammatory disease is
inflammatory bowel disease (e.g., Crohn's disease or ulcerative
colitis).
[0233] In other embodiments, the inflammatory disease is an
autoimmune disease. The autoimmune disease in some embodiments is
rheumatoid arthritis, rheumatic fever, ulcerative colitis, Crohn's
disease, autoimmune inflammatory bowel disease, insulin-dependent
diabetes mellitus, diabetes mellitus, juvenile diabetes,
spontaneous autoimmune diabetes, gastritis, autoimmune atrophic
gastritis, autoimmune hepatitis, thyroiditis, Hashimoto's
thyroiditis, insulitis, oophoritis, orchitis, uveitis, phacogenic
uveitis, multiple sclerosis, myasthenia gravis, primary myxoedema,
thyrotoxicosis, pernicious anemia, autoimmune haemolytic anemia,
Addison's disease, Anklosing spondylitis, sarcoidosis, scleroderma,
Goodpasture's syndrome, Guillain-Barre syndrome, Graves' disease,
glomerulonephritis, psoriasis, pemphigus vulgaris, pemphigoid,
excema, bulous pemiphigous, sympathetic opthalmia, idiopathic
thrombocylopenic purpura, idiopathic feucopenia, Sjogren's
syndrome, systemic sclerosis, Wegener's granulomatosis,
poly/dermatomyositis, primary biliary cirrhosis, primary sclerosing
cholangitis, lupus or systemic lupus erythematosus.
[0234] Graft versus host disease (GVHD) is a complication that can
occur after a pluripotent cell (e.g., stem cell) or bone marrow
transplant in which the newly transplanted material results in an
attack on the transplant recipient's body. In some instances, GVHD
takes place after a blood transfusion. Graft-versus-host-disease
can be divided into acute and chronic forms. The acute or fulminant
form of the disease (aGVHD) is normally observed within the first
100 days post-transplant, and is a major challenge to transplants
owing to associated morbidity and mortality. The chronic form of
graft-versus-host-disease (cGVHD) normally occurs after 100 days.
The appearance of moderate to severe cases of cGVHD adversely
influences long-term survival.
EXAMPLES
Example 1
Immune Response of Synthetic Nanocarriers with Coupled Rapamycin
with and without Ovalbumin Peptide (323-339)
Materials
[0235] Ovalbumin peptide 323-339, a 17 amino acid peptide known to
be a T and B cell epitope of Ovalbumin protein, was purchased from
Bachem Americas Inc. (3132 Kashiwa Street, Torrance Calif. 90505;
Part #4065609). Rapamycin was purchased from TSZ CHEM (185 Wilson
Street, Framingham, Mass. 01702; Product Catalogue # R1017). PLGA
with a lactide:glycolide ratio of 3:1 and an inherent viscosity of
0.75 dL/g was purchased from SurModics Pharmaceuticals (756 Tom
Martin Drive, Birmingham, Ala. 35211; Product Code 7525 DLG 7A).
Polyvinyl alcohol (85-89% hydrolyzed) was purchased from EMD
Chemicals (Product Number 1.41350.1001).
[0236] Solution 1: Ovalbumin peptide 323-339 @ 20 mg/mL in dilute
hydrochloric acid aqueous solution. The solution was prepared by
dissolving ovalbumin peptide in 0.13 M hydrochloric acid solution
at room temperature.
[0237] Solution 2: Rapamycin @ 50 mg/mL in methylene chloride. The
solution was prepared by dissolving rapamycin in pure methylene
chloride.
[0238] Solution 3: PLGA @ 100 mg/mL in methylene chloride. The
solution was prepared by dissolving PLGA in pure methylene
chloride.
[0239] Solution 4: Polyvinyl alcohol @ 50 mg/mL in 100 mM pH 8
phosphate buffer.
Method for Preparing Synthetic Nanocarrier Containing Rapamycin and
Ovalbumin (323-339)
[0240] A primary water-in-oil emulsion was prepared first. W1/O1
was prepared by combining solution 1 (0.2 mL), solution 2 (0.2 mL),
and solution 3 (1.0 mL) in a small pressure tube and sonicating at
50% amplitude for 40 seconds using a Branson Digital Sonifier 250.
A secondary emulsion (W1/O1/W2) was then prepared by combining
solution 4 (3.0 mL) with the primary W1/O1 emulsion, vortexing for
10 s, and sonicating at 30% amplitude for 60 seconds using the
Branson Digital Sonifier 250.
[0241] The W1/O1/W2 emulsion was added to a beaker containing 70 mM
pH 8 phosphate buffer solution (30 mL) and stirred at room
temperature for 2 hours to allow the methylene chloride to
evaporate and for the synthetic nanocarriers to form. A portion of
the synthetic nanocarriers were washed by transferring the
synthetic nanocarrier suspension to a centrifuge tube and
centrifuging at 21,000.times.g and 4.degree. C. for one hour,
removing the supernatant, and re-suspending the pellet in phosphate
buffered saline. The washing procedure was repeated, and the pellet
was re-suspended in phosphate buffered saline for a final synthetic
nanocarrier dispersion of about 10 mg/mL.
[0242] The amounts of peptide and rapamycin in the synthetic
nanocarriers were determined by HPLC analysis. The total
dry-synthetic nanocarrier mass per mL of suspension was determined
by a gravimetric method.
Method for Synthetic Nanocarrier Containing Rapamycin
[0243] A primary water-in-oil emulsion was prepared first. W1/O1
was prepared by combining 0.13 M hydrochloric acid solution (0.2
mL), solution 2 (0.2 mL), and solution 3 (1.0 mL) in a small
pressure tube and sonicating at 50% amplitude for 40 seconds using
a Branson Digital Sonifier 250. A secondary emulsion (W1/O1/W2) was
then prepared by combining solution 4 (3.0 mL) with the primary
W1/O1 emulsion, vortexing for 10 s, and sonicating at 30% amplitude
for 60 seconds using the Branson Digital Sonifier 250.
[0244] The W1/O1/W2 emulsion was added to a beaker containing 70 mM
pH 8 phosphate buffer solution (30 mL) and stirred at room
temperature for 2 hours to allow the methylene chloride to
evaporate and for the synthetic nanocarriers to form. A portion of
the synthetic nanocarriers were washed by transferring the
synthetic nanocarrier suspension to a centrifuge tube and
centrifuging at 21,000.times.g and 4.degree. C. for one hour,
removing the supernatant, and re-suspending the pellet in phosphate
buffered saline. The washing procedure was repeated, and the pellet
was re-suspended in phosphate buffered saline for a final synthetic
nanocarrier dispersion of about 10 mg/mL.
[0245] The amount of rapamycin in the synthetic nanocarrier was
determined by HPLC analysis. The total dry-synthetic nanocarrier
mass per mL of suspension was determined by a gravimetric
method.
Method for Measuring Rapamycin Load
[0246] Approximately 3 mg of synthetic nanocarriers were collected
and centrifuged to separate supernatant from synthetic nanocarrier
pellet. Acetonitrile was added to the pellet, and the sample was
sonicated and centrifuged to remove any insoluble material. The
supernatant and pellet were injected on RP-HPLC and absorbance was
read at 278 nm. The .mu.g found in the pellet were used to
calculate % entrapped (load), .mu.g in supernatant and pellet were
used to calculate total .mu.g recovered.
[0247] Method for Measuring Ovalbumin (323-339) Load
[0248] Approximately 3 mg of synthetic nanocarriers were collected
and centrifuged to separate supernatant from synthetic nanocarrier
pellet. Trifluoroethanol was added to the pellet and the sample was
sonicated to dissolve the polymer, 0.2% trifluoroacetic acid was
added and sample was sonicated and then centrifuged to remove any
insoluble material. The supernatant and pellet were injected on
RP-HPLC and absorbance was read at 215 nm. The .mu.g found in the
pellet were used to calculate % entrapped (load), .mu.g in
supernatant and pellet were used to calculate total .mu.g
recovered.
Antigen-Specific Tolerogenic Dendritic Cells (Tdc) Activity on Treg
Cell Development
[0249] The assay included the use of OTII mice which have a
transgenic T-cell receptor specific for an immune-dominant
ovalbumin (323-339). In order to create antigen-specific tDCs,
CD11c+ splenocytes were isolated, and the ovalbumin (323-339)
peptide added in vitro at 1 .mu.g/ml or no antigen. Soluble or
nanocarrier-encapsulated rapamycin was then added to the DCs for 2
hours which were then washed extensively to remove free rapamycin
from the culture. Purified responder CD4+CD25- cells were isolated
from OTII mice and added to tDC at a 10:1 T to DC ratio. The
mixture of tDC and OTII T-cells were then cultured for 4-5 days,
and the frequency of Treg cells (CD4+CD25highFoxP3+) were analyzed
by flow cytometry as shown in FIG. 1. Regions were selected based
on isotype controls.
Example 2
Mesoporous Silica Nanoparticles with Coupled Ibuprofen
(Prophetic)
[0250] Mesoporous SiO2 nanoparticle cores are created through a
sol-gel process. Hexadecyltrimethyl-ammonium bromide (CTAB) (0.5 g)
is dissolved in deionized water (500 mL), and then 2 M aqueous NaOH
solution (3.5 mL) is added to the CTAB solution. The solution is
stirred for 30 min, and then Tetraethoxysilane (TEOS) (2.5 mL) is
added to the solution. The resulting gel is stirred for 3 h at a
temperature of 80.degree. C. The white precipitate which forms is
captured by filtration, followed by washing with deionized water
and drying at room temperature. The remaining surfactant is then
extracted from the particles by suspension in an ethanolic solution
of HCl overnight. The particles are washed with ethanol,
centrifuged, and redispersed under ultrasonication. This wash
procedure is repeated two additional times.
[0251] The SiO2 nanoparticles are then functionalized with amino
groups using (3-aminopropyl)-triethoxysilane (APTMS). To do this,
the particles are suspended in ethanol (30 mL), and APTMS (50
.mu.L) is added to the suspension. The suspension is allowed to
stand at room temperature for 2 h and then is boiled for 4 h,
keeping the volume constant by periodically adding ethanol.
Remaining reactants are removed by five cycles of washing by
centrifugation and redispersing in pure ethanol.
[0252] In a separate reaction, 1-4 nm diameter gold seeds are
created. All water used in this reaction is first deionized and
then distilled from glass. Water (45.5 mL) is added to a 100 mL
round-bottom flask. While stirring, 0.2 M aqueous NaOH (1.5 mL) is
added, followed by a 1% aqueous solution of
tetrakis(hydroxymethyl)phosphonium chloride (THPC) (1.0 mL). Two
minutes after the addition of THPC solution, a 10 mg/mL aqueous
solution of chloroauric acid (2 mL), which has been aged at least
15 min, is added. The gold seeds are purified through dialysis
against water.
[0253] To form the core-shell nanocarriers, the
amino-functionalized SiO2 nanoparticles formed above are first
mixed with the gold seeds for 2 h at room temperature. The
gold-decorated SiO2 particles are collected through centrifugation
and mixed with an aqueous solution of chloroauric acid and
potassium bicarbonate to form the gold shell. The particles are
then washed by centrifugation and redispersed in water. Ibuprofen
is loaded by suspending the particles in a solution of sodium
ibuprofen (1 mg/L) for 72 h. Free ibuprofen is then washed from the
particles by centrifugation and redispersing in water.
Example 3
Liposomes Containing Cyclosporine A (Prophetic)
[0254] The liposomes are formed using thin film hydration.
1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) (32 .mu.mol),
cholesterol (32 .mu.mol), and cyclosporin A (6.4 .mu.mol) are
dissolved in pure chloroform (3 mL). This lipid solution is added
to a 50 mL round-bottom flask, and the solvent is evaporated on a
rotary evaporator at a temperature of 60.degree. C. The flask is
then flushed with nitrogen gas to remove remaining solvent.
Phosphate buffered saline (2 mL) and five glass beads are added to
the flask, and the lipid film is hydrated by shaking at 60.degree.
C. for 1 h to form a suspension. The suspension is transferred to a
small pressure tube and sonicated at 60.degree. C. for four cycles
of 30s pulses with a 30 s delay between each pulse. The suspension
is then left undisturbed at room temperature for 2 h to allow for
complete hydration. The liposomes are washed by centrifugation
followed by resuspension in fresh phosphate buffered saline.
Example 4
Polymeric Nanocarrier Containing Polymer-Rapamycin Conjugate
(Prophetic)
[0255] Preparation of PLGA-Rapamycin Conjugate:
[0256] PLGA polymer with acid end group (7525 DLG1A, acid number
0.46 mmol/g, Lakeshore Biomaterials; 5 g, 2.3 mmol, 1.0 eq) is
dissolved in 30 mL of dichloromethane (DCM).
N,N-Dicyclohexylcarbodimide (1.2 eq, 2.8 mmol, 0.57 g) is added
followed by rapamycin (1.0 eq, 2.3 mmol, 2.1 g) and
4-dimethylaminopyridine (DMAP) (2.0 eq, 4.6 mmol, 0.56 g). The
mixture is stirred at rt for 2 days. The mixture is then filtered
to remove insoluble dicyclohexylurea. The filtrate is concentrated
to ca. 10 mL in volume and added to 100 mL of isopropyl alcohol
(IPA) to precipitate out the PLGA-rapamycin conjugate. The IPA
layer is removed and the polymer is then washed with 50 mL of IPA
and 50 mL of methyl t-butyl ether (MTBE). The polymer is then dried
under vacuum at 35 C for 2 days to give PLGA-rapamycin as a white
solid (ca. 6.5 g).
[0257] Preparation of nanocarrier containing PLGA-rapamycin
conjugate and ovalbumin peptide (323-339):
[0258] Nanocarrier containing PLGA-rapamycin is prepared according
to the procedure described in Example 1 as follows:
[0259] Solutions for nanocarrier formation are prepared as
follows:
[0260] Solution 1: Ovalbumin peptide 323-339 @ 20 mg/mL in dilute
hydrochloric acid aqueous solution. The solution is prepared by
dissolving ovalbumin peptide in 0.13 M hydrochloric acid solution
at room temperature. Solution 2: PLGA-rapamycin @ 100 mg/mL in
methylene chloride. The solution is prepared by dissolving
PLGA-rapamycin in pure methylene chloride. Solution 3: PLA-PEG @
100 mg/mL in methylene chloride. The solution is prepared by
dissolving PLA-PEG in pure methylene chloride. Solution 4:
Polyvinyl alcohol @ 50 mg/mL in 100 mM pH 8 phosphate buffer.
[0261] A primary water-in-oil emulsion is prepared first. W1/O1 is
prepared by combining solution 1 (0.2 mL), solution 2 (0.75 mL),
and solution 3 (0.25 mL) in a small pressure tube and sonicating at
50% amplitude for 40 seconds using a Branson Digital Sonifier 250.
A secondary emulsion (W1/O1/W2) is then prepared by combining
solution 4 (3.0 mL) with the primary W1/O1 emulsion, vortexing for
10 s, and sonicating at 30% amplitude for 60 seconds using the
Branson Digital Sonifier 250. The W1/O1/W2 emulsion is added to a
beaker containing 70 mM pH 8 phosphate buffer solution (30 mL) and
stirred at room temperature for 2 hours to allow the methylene
chloride to evaporate and for the nanocarriers to form. A portion
of the nanocarriers is washed by transferring the nanocarrier
suspension to a centrifuge tube and centrifuging at 75,600.times.g
and 4.degree. C. for 35 min, removing the supernatant, and
re-suspending the pellet in phosphate buffered saline. The washing
procedure is repeated, and the pellet is re-suspended in phosphate
buffered saline for a final nanocarrier dispersion of about 10
mg/mL.
Example 5
Preparation of Gold Nanocarriers (AuNCs) Containing Rapamycin
(Prophetic)
[0262] Preparation of HS-PEG-Rapamycin:
[0263] A solution of PEG acid disulfide (1.0 eq), rapamycin
(2.0-2.5 eq), DCC (2.5 eq) and DMAP (3.0 eq) in dry DMF is stirred
at rt overnight. The insoluble dicyclohexylurea is removed by
filtration and the filtrate is added to isopropyl alcohol (IPA) to
precipitate out the PEG-disulfide-di-rapamycin ester and washed
with IPA and dried. The polymer is then treated with
tris(2-carboxyethyl)phosphine hydrochloride in DMF to reduce the
PEG disulfide to thiol PEG rapamycin ester (HS-PEG-rapamycin). The
resulting polymer is recovered by precipitation from IPA and dried
as previously described and analyzed by H NMR and GPC.
[0264] Formation of Gold NCs (AuNCs):
[0265] An aq. solution of 500 mL of 1 mM HAuCl4 is heated to reflux
for 10 min with vigorous stirring in a 1 L round-bottom flask
equipped with a condenser. A solution of 50 mL of 40 mM of
trisodium citrate is then rapidly added to the stirring solution.
The resulting deep wine red solution is kept at reflux for 25-30
min and the heat is withdrawn and the solution is cooled to room
temperature. The solution is then filtered through a 0.8 .mu.m
membrane filter to give the AuNCs solution. The AuNCs are
characterized using visible spectroscopy and transmission electron
microscopy. The AuNCs are ca. 20 nm diameter capped by citrate with
peak absorption at 520 nm.
[0266] AuNCs conjugate with HS-PEG-rapamycin:
[0267] A solution of 150 .mu.l of HS-PEG-rapamycin (10 .mu.M in 10
mM pH 9.0 carbonate buffer) is added to 1 mL of 20 nm diameter
citrate-capped gold nanocarriers (1.16 nM) to produce a molar ratio
of thiol to gold of 2500:1. The mixture is stirred at room
temperature under argon for 1 hour to allow complete exchange of
thiol with citrate on the gold nanocarriers. The AuNCs with
PEG-rapamycin on the surface is then purified by centrifuge at
12,000 g for 30 minutes. The supernatant is decanted and the pellet
containing AuNC-S-PEG-rapamycin is then pellet washed with
1.times.PBS buffer. The purified Gold-PEG-rapamycin nanocarriers
are then resuspend in suitable buffer for further analysis and
bioassays.
Example 6
Mesoporous Silica-gold Core-shell Nanocarriers Containing Ovalbumin
(Prophetic)
[0268] Mesoporous SiO2 nanoparticle cores are created through a
sol-gel process. Hexadecyltrimethyl-ammonium bromide (CTAB) (0.5 g)
is dissolved in deionized water (500 mL), and then 2 M aqueous NaOH
solution (3.5 mL) is added to the CTAB solution. The solution is
stirred for 30 min, and then Tetraethoxysilane (TEOS) (2.5 mL) is
added to the solution. The resulting gel is stirred for 3 h at a
temperature of 80.degree. C. The white precipitate which forms is
captured by filtration, followed by washing with deionized water
and drying at room temperature. The remaining surfactant is then
extracted from the particles by suspension in an ethanolic solution
of HCl overnight. The particles are washed with ethanol,
centrifuged, and redispersed under ultrasonication. This wash
procedure is repeated two additional times.
[0269] The SiO2 nanoparticles are then functionalized with amino
groups using (3-aminopropyl)-triethoxysilane (APTMS). To do this,
the particles are suspended in ethanol (30 mL), and APTMS (50
.mu.L) is added to the suspension. The suspension is allowed to
stand at room temperature for 2 h and then is boiled for 4 h,
keeping the volume constant by periodically adding ethanol.
Remaining reactants are removed by five cycles of washing by
centrifugation and redispersing in pure ethanol.
[0270] In a separate reaction, 1-4 nm diameter gold seeds are
created. All water used in this reaction is first deionized and
then distilled from glass. Water (45.5 mL) is added to a 100 mL
round-bottom flask. While stirring, 0.2 M aqueous NaOH (1.5 mL) is
added, followed by a 1% aqueous solution of
tetrakis(hydroxymethyl)phosphonium chloride (THPC) (1.0 mL). Two
minutes after the addition of THPC solution, a 10 mg/mL aqueous
solution of chloroauric acid (2 mL), which has been aged at least
15 min, is added. The gold seeds are purified through dialysis
against water.
[0271] To form the core-shell nanocarriers, the
amino-functionalized SiO2 nanoparticles formed above are first
mixed with the gold seeds for 2 h at room temperature. The
gold-decorated SiO2 particles are collected through centrifugation
and mixed with an aqueous solution of chloroauric acid and
potassium bicarbonate to form the gold shell. The particles are
then washed by centrifugation and redispersed in water. Thiolated
Ovalbumin (made by treating Ovalbumin with 2-iminothiolane
hydrochloride) is loaded by suspending the particles in a solution
of thiolated Ovalbumin (1 mg/L) for 72 h. The particles is then
pellet washed with 1.times.PBS (pH 7.4) to remove free protein. The
resulting silica-gold core-shell nanocarriers containing Ovalbumin
are then re-suspended in 1.times.PBS for further analysis and
assays.
Example 7
Liposomes Containing Rapamycin and Ovalbumin (Prophetic)
[0272] The liposomes are formed by thin film hydration.
1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) (32 .mu.mol),
cholesterol (32 .mu.mol), and rapamycin (6.4 .mu.mol) are dissolved
in pure chloroform (3 mL). This lipid solution is added to a 10 mL
glass tube and the solvent is removed under nitrogen gas stream and
desiccated for 6 hr. under vacuum. Multilamellar vesicles are
obtained by hydration of the film with 2.0 ml of 25 mM MOPS buffer
pH 8.5, containing excess amount of Ovalbumin. The tube is vortexed
until the lipid film is peeled of from the tube surface. To break
the multilamellar vesicles into monolamellar, ten cycles of
freezing (liquid nitrogen) and thawing (30.degree. C. water bath)
are applied. The sample is then diluted to 1 ml in 25 mM MOPS
buffer pH 8.5. Size of the resulting liposome is homogenized by
extrusion by passing the sample 10 fold through a 200 nm pore
polycarbonate filters. The resulting liposomes are then used for
further analysis and bioassays.
Example 8
Polymeric Nanocarriers Composed of Modified Polyamino Acid with
Surface Conjugated Ovalbumin (Prophetic)
[0273] Step-1. Preparation of Poly(.gamma.-glutamic acid)
(.gamma.-PGA) modified with L-phenylalanine ethyl ester (L-PAE):
4.7 unit mmol of .gamma.-PGA (Mn=300 kD) is dissolved in 0.3
N-NaHCO3 aqueous solution (50 mL). L-PAE (4.7 mmol) and EDC.HCl
(4.7 mmol) are added to the solution and stirred for 30 min at 4 C.
The solution is then maintained at room temperature with stirring
for 24 h. Low-molecular-weight chemicals are removed by dialysis
using dialysis membrane with MWCO 50 kD. The resulting
.gamma.-PGA-graft-L-PAE is obtained by freeze-drying.
[0274] Step-2. Preparation of nanoparticles from
.gamma.-PGA-graft-L-PAE polymer: Nanoparticles composed of
.gamma.-PGA-graft-L-PAE are prepared by a precipitation and
dialysis method. .gamma.-PGA-graft-L-PAE (20 mg) was dissolved in 2
ml of DMSO followed by addition of 2 mL of water to form a
translucent solution. The solution is then dialyzed against
distilled water using cellulose membrane tubing (50,000 MWCO) to
form the nanoparticles and to remove the organic solvents for 72 h
at room temperature. The distilled water is exchanged at intervals
of 12 h. The resulting nanoparticle solution (10 mg/mL in water) is
then used for antigen conjugation.
[0275] Step-3. Ovalbumin conjugation to .gamma.-PGA nanoparticles:
Surface carboxylic acid groups of the .gamma.-PGA nanoparticles (10
mg/ml) are first activated by EDC and NHS (10 mg/mL each in
phosphate buffer, pH 5.8) for 2 h at ambient temperature. After
pellet washing to remove excess EDC/NHS, the activated
nanoparticles are mixed with 1 mL of Ovalbumin (10 mg/ml) in
phosphate-buffered saline (PBS, pH 7.4) and the mixture is
incubated at 4-8 C for 24 h. The resulting Ovalbumin conjugated
7-PGA nanoparticles are washed twice with PBS and resuspended at 5
mg/mL in PBS for further analysis and bioassays.
Example 9
Erythropoietin (EPO)-Encapsulated .gamma.-PGA Nanoparticles
(Prophetic)
[0276] To prepare the EPO-encapsulated .gamma.-PGA nanoparticles,
0.25-4 mg of EPO is dissolved in 1 mL of PBS (pH 7.4) and 1 mL of
the .gamma.-PGA-graft-L-PAE (10 mg/mL in DMSO) is added to the EPO
solution. The resulting solution is centrifuged at 14,000.times.g
for 15 min and repeatedly rinsed with PBS. The resulting
EPO-encapsulated .gamma.-PGA nanoparticles are then resuspended in
PBS (5 mg/mL) for further analysis and bioassay.
Example 10
Preparation of Gold Nanocarriers (AuNCs) Containing Ovalbumin
(Prophetic)
[0277] Step-1. Formation of Gold NCs (AuNCs): An aq. solution of
500 mL of 1 mM HAuCl4 is heated to reflux for 10 min with vigorous
stirring in a 1 L round-bottom flask equipped with a condenser. A
solution of 50 mL of 40 mM of trisodium citrate is then rapidly
added to the stirring solution. The resulting deep wine red
solution is kept at reflux for 25-30 min and the heat is withdrawn
and the solution is cooled to room temperature. The solution is
then filtered through a 0.8 .mu.m membrane filter to give the AuNCs
solution. The AuNCs are characterized using visible spectroscopy
and transmission electron microscopy. The AuNCs are ca. 20 nm
diameter capped by citrate with peak absorption at 520 nm.
[0278] Step-2. Conjugation of Ovalbumin to AuNCs: A solution of 150
.mu.l of thiolated Ovalbumin (10 .mu.M in 10 mM pH 9.0 carbonate
buffer) is added to 1 mL of 20 nm diameter citrate-capped gold
nanocarriers (1.16 nM) to produce a molar ratio of thiol to gold of
2500:1. The mixture is stirred at room temperature under argon for
1 hour to allow complete exchange of thiol with citrate on the gold
nanocarriers. The AuNCs with Ovalbumin on the surface is then
purified by centrifuge at 12,000 g for 30 minutes. The supernatant
is decanted and the pellet containing AuNC-Ovalbumin is then pellet
washed with 1.times.PBS buffer. The purified Gold-Ovalbumin
nanocarriers are then resuspend in suitable buffer for further
analysis and bioassays.
Example 11
Evaluating Tolerogenic Immune Response to Antigen In Vivo
(Prophetic)
[0279] Balb/c mice are immunized with an antigen in incomplete
Freund's adjuvant, and the level of regulatory B cell proliferation
is assessed. Subsequently, a composition of the invention is
administered in a dose-dependent manner. The same mice are then
again exposed to the antigen, and the level of regulatory B cell
proliferation is again assessed. Changes in regulatory B cell
proliferation are then monitored with an increase upon subsequent
challenge with the antigen indicating a tolerogenic immune
response.
Example 12
Evaluating In Vitro Induction of Regulatory B Cells (Prophetic)
[0280] A cell population comprising regulatory B cells or
regulatory B cell precursors is contacted in vitro with a
composition provided herein. After a time sufficient for the
composition to interact with the regulatory B cells or regulatory B
cell precursors in the cell population, an increase in the number
of regulatory B cells is expected. A time sufficient for the
increase in the number of regulatory B cells in the population of
cells is, in some embodiment, a period of about 1 day, about 2
days, about 3 days, about 4 days, about 5 days, about 6 days, about
1 week, about 2 weeks, about 3 weeks, or about 4 weeks. In some
embodiments, the number of regulatory B cells is increased after
that time, for example, to at least about 5 times, at least about
10 times, at least about 20 times, at least about 50 times, at
least about 100 times, at least about 1.000 times, at least about
10,000 times, at least about 100,000 times, or at least about
1,000,000 times as compared to the original total or relative
number of regulatory B cells in the population. In some
embodiments, the number of regulatory B cells is increased after
that time, for example, to about 1%, about 5%, about 10%, about
15%, about 20%, about 25%, about 30%, about 40%, about 50%, about
75%, about 90%, or about 95% of the total cells in the population
or of the lymphocytes in the cell population. In some embodiments,
regulatory B cells are absent from the cell population before the
contacting, but are present after the contacting.
[0281] In some embodiments, the total and/or relative number of
regulatory B cells in the population is determined before the
population of cells is contacted with a composition provided herein
to establish a baseline number of regulatory B cells in the
population. In some embodiments, the population of cells is
contacted once with a composition provided herein. In some
embodiments, the population of cells is contacted repeatedly with a
composition provided herein.
[0282] The number and/or presence of regulatory B cells in the
population of cells is also determined after the contacting. In
some embodiments, the number and/or presence of regulatory B cells
is monitored over a period of time, for example, by performing a
plurality of subsequent regulatory B cell detection assays. In some
embodiments, the number and/or presence of regulatory B cells in
the population of cells is determined by taking a sample from the
cell population that is representative of the cell population,
staining cells contained in that sample with antibodies or staining
agents that specifically detect regulatory B cell markers, and
detecting cells that express regulatory B cell markers in the
sample, for example, by FACS or by immunohistochemistry. In some
embodiments, the regulatory B cells are quantified. In some
embodiments, the quantity of regulatory B cells determined after
the contacting is compared to the quantity of regulatory B cells
before the contacting, for example, to the baseline number of
regulatory B cells, wherein, if the number of regulatory B cells in
the population of cells is higher after the contacting than the
baseline number, then it is determined that a tolerogenic response
to the composition has occurred.
Example 13
Evaluating In Vivo Induction of Regulatory B Cells (Prophetic)
[0283] A cell population comprising regulatory B cells or
regulatory B cell precursors is contacted in vivo with a
composition provided herein. In some embodiments, the composition
is administered to a subject harboring the cell population
comprising regulatory B cells or regulatory B cell precursors.
After a time sufficient for the composition to interact with the
regulatory B cells or regulatory B cell precursors in the subject,
an increase in the number of regulatory B cells is expected. A time
sufficient for the increase in the number of regulatory B cells in
the population is, in some embodiment, a period of about 1 day,
about 2 days, about 3 days, about 4 days, about 5 days, about 6
days, about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks.
In some embodiments, the period of time sufficient to effect an
increase in the number of regulatory B cells is longer than 4
weeks. In some embodiments, the number of regulatory B cells is
increased after that time to at least about 5 times, at least about
10 times, at least about 20 times, at least about 50 times, at
least about 100 times, at least about 1.000 times, at least about
10,000 times, at least about 100,000 times, or at least about
1,000,000 times as compared to the original total or relative
number of regulatory B cells in the subject. In some embodiments,
the number of regulatory B cells is increased after that time, for
example, to about 1%, about 5%, about 10%, about 15%, about 20%,
about 25%, about 30%, about 40%, about 50%, about 75%, about 90%,
or about 95% of the total cells in the subject, or of a lymphocyte
population in the subject. In some embodiments, regulatory B cells
are absent from the subject before the administration, but are
present after the administration.
[0284] In some embodiments, the total and/or relative number of
regulatory B cells in the subject is determined before the subject
is administered a composition provided herein to establish a
baseline number of regulatory B cells in the subject. In some
embodiments, the composition provided herein is administered to a
subject once. In some embodiments, a composition provided herein is
administered to a subject multiple times, for example, until a
desired increase or abundance of regulatory B cells is observed in
the subject.
[0285] The number and/or presence of regulatory B cells in the
subject is determined after the administration. In some
embodiments, the number and/or presence of regulatory B cells is
monitored over a period of time, for example, by performing a
plurality of subsequent regulatory B cell detection assays. In some
embodiments, the number and/or presence of regulatory B cells in
the subject is determined by taking a sample from the subject, for
example, a peripheral blood sample, or a lymph sample, that is
representative of a lymphocyte population in the subject, staining
cells contained in that sample with antibodies or staining agents
that specifically detect regulatory B cell markers, and detecting
cells that express regulatory B cell markers in the sample, for
example, by FACS or by immunohistochemistry. In some embodiments,
the regulatory B cells are quantified. In some embodiments, the
quantity of regulatory B cells determined after administration is
compared to the quantity of regulatory B cells before
administration, for example, to the baseline number of regulatory B
cells, wherein, if the number of regulatory B cells in the subject
is higher after the administration than the baseline number, then
it is determined that a tolerogenic response to the composition has
occurred.
Example 14
In Vivo Induction of a Desired Immune Response Against
Transplantable Bone Marrow Cells (Prophetic)
[0286] A population of at least 10.sup.6 synthetic nanocarriers
comprising immunosuppressant and antigens obtained or derived from
bone marrow cells are administered subcutaneously to a subject four
weeks prior to the subject receiving a bone marrow transplant.
After the transplant is received by the subject, the generation of
a desired immune response in the subject is assessed once daily
during the first week after receiving the transplant, and then
weekly for the next three weeks, and then monthly for the next 11
months. As part of the assessment, regulatory B cell counts are
taken and compared to regulatory B cell counts taken prior to
administering the bone marrow transplant or the synthetic
nanocarriers to the subject. During the first year, maintenance
doses of the synthetic nanocarriers are administered bi-monthly to
the subject. The subject is expected to exhibit higher or desired
levels of regulatory B cells specific to the bone marrow
transplant.
Example 15
Assessing the Effects of Nanocarriers with Antigens and
Immunosuppressants
Nanocarriers
[0287] Ovalbumin peptide 323-339, a 17 amino acid peptide known to
be a T and B cell epitope of Ovalbumin protein, was purchased from
Bachem Americas Inc. (3132 Kashiwa Street, Torrance Calif. 90505;
Part #4065609). PLGA with a lactide:glycolide ratio of 3:1 and an
inherent viscosity of 0.75 dL/g was purchased from SurModics
Pharmaceuticals (756 Tom Martin Drive, Birmingham, Ala. 35211;
Product Code 7525 DLG 7A). PLA-PEG block co-polymer with a PEG
block of approximately 5,000 Da and PLA block of approximately
20,000 Da was synthesized. Polyvinyl alcohol (85-89% hydrolyzed)
was purchased from EMD Chemicals (Product Number 1.41350.1001).
[0288] Solutions were prepared as follows: Solution 1: Ovalbumin
peptide 323-339 @ 20 mg/mL in dilute hydrochloric acid aqueous
solution. The solution was prepared by dissolving ovalbumin peptide
in 0.13 M hydrochloric acid solution at room temperature. Solution
2: PLGA @ 100 mg/mL in methylene chloride. The solution was
prepared by dissolving PLGA in pure methylene chloride. Solution 3:
PLA-PEG @ 100 mg/mL in methylene chloride. The solution was
prepared by dissolving PLA-PEG in pure methylene chloride. Solution
4: Polyvinyl alcohol @ 50 mg/mL in 100 mM pH 8 phosphate buffer. A
primary water-in-oil emulsion was prepared first. W1/O1 was
prepared by combining solution 1 (0.2 mL), solution 2 (0.75 mL),
and solution 3 (0.25 mL) in a small pressure tube and sonicating at
50% amplitude for 40 seconds using a Branson Digital Sonifier 250.
A secondary emulsion (W1/O1/W2) was then prepared by combining
solution 4 (3.0 mL) with the primary W1/O1 emulsion, vortexing for
10 s, and sonicating at 30% amplitude for 60 seconds using the
Branson Digital Sonifier 250.
[0289] The W1/O1/W2 emulsion was added to a beaker containing 70 mM
pH 8 phosphate buffer solution (30 mL) and stirred at room
temperature for 2 hours to allow the methylene chloride to
evaporate and for the nanocarriers to form. A portion of the
nanocarriers were washed by transferring the nanocarrier suspension
to a centrifuge tube and centrifuging at 75,600.times.g and
4.degree. C. for 35 min, removing the supernatant, and
re-suspending the pellet in phosphate buffered saline. The washing
procedure was repeated, and the pellet was re-suspended in
phosphate buffered saline for a final nanocarrier dispersion of
about 10 mg/mL.
[0290] Nanocarrier size was determined by dynamic light scattering.
The amount of peptide in the nanocarrier was determined by HPLC
analysis. The total dry-nanocarrier mass per mL of suspension was
determined by a gravimetric method.
TABLE-US-00002 Effective Diameter Peptide Content Nanocarrier (nm)
(% w/w) 1 234 2.1
[0291] Ovalbumin peptide 323-339, a 17 amino acid peptide known to
be a T and B cell epitope of Ovalbumin protein, was purchased from
Bachem Americas Inc. (3132 Kashiwa Street, Torrance Calif. 90505;
Part #4065609). Rapamycin was purchased from TSZ CHEM (185 Wilson
Street, Framingham, Mass. 01702; Product Catalogue # R1017). PLGA
with a lactide:glycolide ratio of 3:1 and an inherent viscosity of
0.75 dL/g was purchased from SurModics Pharmaceuticals (756 Tom
Martin Drive, Birmingham, Ala. 35211; Product Code 7525 DLG 7A).
PLA-PEG block co-polymer with a PEG block of approximately 5,000 Da
and PLA block of approximately 20,000 Da was synthesized. Polyvinyl
alcohol (85-89% hydrolyzed) was purchased from EMD Chemicals
(Product Number 1.41350.1001).
[0292] Solutions were prepared as follows: Solution 1: Ovalbumin
peptide 323-339 @ 20 mg/mL in dilute hydrochloric acid aqueous
solution. The solution was prepared by dissolving ovalbumin peptide
in 0.13 M hydrochloric acid solution at room temperature. Solution
2: Rapamycin @ 50 mg/mL in methylene chloride. The solution was
prepared by dissolving rapamycin in pure methylene chloride.
Solution 3: PLGA @ 100 mg/mL in methylene chloride. The solution
was prepared by dissolving PLGA in pure methylene chloride.
Solution 4: PLA-PEG @ 100 mg/mL in methylene chloride. The solution
was prepared by dissolving PLA-PEG in pure methylene chloride.
Solution 5: Polyvinyl alcohol @ 50 mg/mL in 100 mM pH 8 phosphate
buffer.
[0293] A primary water-in-oil emulsion was prepared first. W1/O1
was prepared by combining solution 1 (0.2 mL), solution 2 (0.2 mL),
solution 3 (0.75 mL), and solution 4 (0.25 mL) in a small pressure
tube and sonicating at 50% amplitude for 40 seconds using a Branson
Digital Sonifier 250. A secondary emulsion (W1/O1/W2) was then
prepared by combining solution 5 (3.0 mL) with the primary W1/O1
emulsion, vortexing for 10 s, and sonicating at 30% amplitude for
60 seconds using the Branson Digital Sonifier 250.
[0294] The W1/O1/W2 emulsion was added to a beaker containing 70 mM
pH 8 phosphate buffer solution (30 mL) and stirred at room
temperature for 2 hours to allow the methylene chloride to
evaporate and for the nanocarriers to form. A portion of the
nanocarriers were washed by transferring the nanocarrier suspension
to a centrifuge tube and centrifuging at 21,000.times.g and
4.degree. C. for 45 min, removing the supernatant, and
re-suspending the pellet in phosphate buffered saline. The washing
procedure was repeated, and the pellet was re-suspended in
phosphate buffered saline for a final nanocarrier dispersion of
about 10 mg/mL.
[0295] Nanocarrier size was determined by dynamic light scattering.
The amounts of peptide and rapamycin in the nanocarrier were
determined by HPLC analysis. The total dry-nanocarrier mass per mL
of suspension was determined by a gravimetric method.
TABLE-US-00003 Effective Rapamycin Peptide Diameter Content Content
Nanocarrier ID (nm) (% w/w) (% w/w) 2 227 9.0 2.5
[0296] Measurement of synthetic nanocarrier dimensions was obtained
by dynamic light scattering (DLS). A suspension of the synthetic
nanocarriers was diluted with purified water to achieve a final
synthetic nanocarrier suspension concentration of approximately
0.01 to 0.1 mg/mL. The diluted suspension was prepared directly
inside a suitable cuvette for DLS analysis. The cuvette was then
placed in a Brookhaven Instruments Corp. ZetaPALS, allowed to
equilibrate to 25.degree. C., and then scanned for sufficient time
to acquire a stable and reproducible distribution based on
appropriate inputs for viscosity of the medium and refractive
indicies of the sample. The effective diameter, or mean of the
distribution, was then reported.
Immunization
[0297] The nanocarriers were thawed and equilibrated. Initial
dilutions constituted a 10.times. stock solution, and were further
diluted to a concentration of 100 .mu.g/ml in OVA.sub.323-339, or a
1.times. solution. This 1.times. solution was used for injections
at 200 .mu.l per i.v. injection. Animals were immunized with OVA
protein (OVA) and treated with OVA.sub.323-339 peptide.
Immunization routes were as follows: 10 .mu.g of OVA+4 mg Alum i.p.
in 400 .mu.l per each Balb/C immunologically naive female mouse.
Experimental groups consisted of 5 animals each. Spleen cells were
restimulated with antigen using CFSE or CTO to determine the amount
of Ag-specific proliferation.
Measuring Regulatory B Cells
[0298] The frequency of Ovalbumin reactive IL-10 secreting B cells
was calculated by way of flow cytometry. Splenocytes from
experimental animals were stained with CFSE, a thiol-reactive
Fluorescent Probe suitable for long-term cell labeling, and
cultured in complete media at 37 C, 5% CO2 with Ovalbumin protein
for 3 days. On day 3 the cells were assayed for their potential to
secrete different cytokines by intracellular staining using
standard methods and kits. Briefly, cells were restimulated with
phorbol myristate acetate (PMA) and lonomycin for 2 hours and
protein transport was blocked for another 4 hours. Unspecific
binding of antibodies was blocked with anti-CD16/32 antibody and
then cells were stained with conjugated antibodies specific
recognizing specifically B cells (CD45R (B220) and CD19). After
fixation with paraformaldehyde cells were permeabilized to allow
monoclonal antibodies into the cells and label intracellular
epitopes (cytokines). Splenocytes that were B220+CD19+ were
assessed for proliferation by comparing the differential CFSE
staining and the proportion of IL-10-secreting cells
determined.
Measurement of IgE Antibodies
[0299] IgE antibodies were measured using a Mouse OVA-IgE ELISA kit
provided by MDBioproducts (Cat# M036005) consistent with the
manufacturer's instructions.
Results
[0300] FIG. 2 demonstrate the effectiveness of the nanocarriers in
the generation of regulatory B cells in lavage samples from animal
subjects treated with synthetic nanocarriers comprising
OVA.sub.323-339 and immunosuppressant. The figure demonstrates that
synthetic nanocarriers of the invention resulted in the production
of IL-10 and TGF-.beta. by antigen-specific CD24+ B cells. FIG. 3
shows a decrease in IgE production with synthetic nanocarriers of
the invention.
Example 16
Isolation of Regulatory B Cell (Bregs) Cells from a Subject after
Administration of Inventive Synthetic Nanocarriers (Prophetic)
[0301] Bregs are isolated from biological samples, for example,
from peripheral blood, obtained from a subject after the subject is
administered a synthetic nanocarrier composition as described
herein. Typically, the biological sample is obtained from the
subject after a time period sufficient for the administered itDCs
to induce Bregs. Bregs are isolated from the biological sample, for
example, from whole blood, by negative and/or positive
selection.
[0302] For example, the cellular fraction of whole blood is
obtained by centrifugation, and erythrocytes are lysed using
erythrocyte lysis buffer. After lysis, peripheral blood mononuclear
cells are depleted for CD4+ cells and CD8+ T cells. Subsequently,
Bregs are enriched for by positive selection for the markers
provided elsewhere herein or otherwise known in the art. In some
embodiments, the isolation of regulatory B cells is performed with
a cocktail of biotinylated antibodies and anti-biotin magnetic
beads for the recognition of one or more of these markers.
Sequence CWU 1
1
943118PRTArtificial SequenceHomo sapiens Aggrecan core protein
precursor epitope 1Ala Gly Met Asp Met Cys Ser Ala Gly Trp Leu Ala
Asp Arg Ser Val 1 5 10 15 Arg Tyr 218PRTArtificial SequenceHomo
sapiens Aggrecan core protein precursor epitope 2Glu Asp Ser Glu
Ala Thr Leu Glu Val Val Val Lys Gly Ile Val Phe 1 5 10 15 His Tyr
318PRTArtificial SequenceHomo sapiens Aggrecan core protein
precursor epitope 3Ser Arg Val Ser Lys Glu Lys Glu Val Val Leu Leu
Val Ala Thr Glu 1 5 10 15 Gly Arg 418PRTArtificial SequenceHomo
sapiens Aggrecan core protein precursor epitope 4Val Val Leu Leu
Val Ala Thr Glu Gly Arg Val Arg Val Asn Ser Ala 1 5 10 15 Tyr Gln
518PRTArtificial SequenceHomo sapiens Aggrecan core protein
precursor epitope 5Val Val Val Lys Gly Ile Val Phe His Tyr Arg Ala
Ile Ser Thr Arg 1 5 10 15 Tyr Thr 69PRTArtificial SequenceHomo
sapiens alpha 2 type VI collagen isoform 2C2 precursor epitope 6Asp
Arg Ala Ser Phe Ile Lys Asn Leu 1 5 720PRTArtificial SequenceHomo
sapiens arrestin epitope 7Ala Ser Ser Thr Ile Ile Lys Glu Gly Ile
Asp Arg Thr Val Leu Gly 1 5 10 15 Ile Leu Val Ser 20
820PRTArtificial SequenceHomo sapiens arrestin epitope 8Ala Ser Thr
Pro Thr Lys Leu Gln Glu Ser Leu Leu Lys Lys Leu Gly 1 5 10 15 Ser
Asn Thr Tyr 20 920PRTArtificial SequenceHomo sapiens arrestin
epitope 9Asp Arg Thr Val Leu Gly Ile Leu Val Ser Tyr Gln Ile Lys
Val Lys 1 5 10 15 Leu Thr Val Ser 20 1020PRTArtificial SequenceHomo
sapiens arrestin epitope 10Glu Phe Ala Arg His Asn Leu Lys Asp Ala
Gly Glu Ala Glu Glu Gly 1 5 10 15 Lys Arg Asp Lys 20
1120PRTArtificial SequenceHomo sapiens arrestin epitope 11Glu Pro
Asn His Val Ile Phe Lys Lys Ile Ser Arg Asp Lys Ser Val 1 5 10 15
Thr Ile Tyr Leu 20 1220PRTArtificial SequenceHomo sapiens arrestin
epitope 12Phe Glu Val Lys Ala Phe Ala Thr Asp Ser Thr Asp Ala Glu
Glu Asp 1 5 10 15 Lys Ile Pro Lys 20 1320PRTArtificial SequenceHomo
sapiens arrestin epitope 13Gly Phe Leu Gly Glu Leu Thr Ser Ser Glu
Val Ala Thr Glu Val Pro 1 5 10 15 Phe Arg Leu Met 20
1420PRTArtificial SequenceHomo sapiens arrestin epitope 14Gly Lys
Ile Lys His Glu Asp Thr Asn Leu Ala Ser Ser Thr Ile Ile 1 5 10 15
Lys Glu Gly Ile 20 1520PRTArtificial SequenceHomo sapiens arrestin
epitope 15Gly Asn Arg Asp Tyr Ile Asp His Val Ser Gln Val Gln Pro
Val Asp 1 5 10 15 Gly Val Val Leu 20 1620PRTArtificial SequenceHomo
sapiens arrestin epitope 16Lys Pro Val Ala Met Glu Glu Ala Gln Glu
Lys Val Pro Pro Asn Ser 1 5 10 15 Thr Leu Thr Lys 20
1720PRTArtificial SequenceHomo sapiens arrestin epitope 17Lys Val
Pro Pro Asn Ser Thr Leu Thr Lys Thr Leu Thr Leu Leu Pro 1 5 10 15
Leu Leu Ala Asn 20 1820PRTArtificial SequenceHomo sapiens arrestin
epitope 18Leu Leu Lys Lys Leu Gly Ser Asn Thr Tyr Pro Phe Leu Leu
Thr Phe 1 5 10 15 Pro Asp Tyr Leu 20 1920PRTArtificial SequenceHomo
sapiens arrestin epitope 19Leu Thr Phe Arg Arg Asp Leu Tyr Phe Ser
Arg Val Gln Val Tyr Pro 1 5 10 15 Pro Val Gly Ala 20
2020PRTArtificial SequenceHomo sapiens arrestin epitope 20Met Ala
Ala Ser Gly Lys Thr Ser Lys Ser Glu Pro Asn His Val Ile 1 5 10 15
Phe Lys Lys Ile 20 2120PRTArtificial SequenceHomo sapiens arrestin
epitope 21Asn Arg Glu Arg Arg Gly Ile Ala Leu Asp Gly Lys Ile Lys
His Glu 1 5 10 15 Asp Thr Asn Leu 20 2220PRTArtificial SequenceHomo
sapiens arrestin epitope 22Pro Cys Ser Val Met Leu Gln Pro Ala Pro
Gln Asp Ser Gly Lys Ser 1 5 10 15 Cys Gly Val Asp 20
2320PRTArtificial SequenceHomo sapiens arrestin epitope 23Pro Phe
Leu Leu Thr Phe Pro Asp Tyr Leu Pro Cys Ser Val Met Leu 1 5 10 15
Gln Pro Ala Pro 20 2420PRTArtificial SequenceHomo sapiens arrestin
epitope 24Gln Asp Ser Gly Lys Ser Cys Gly Val Asp Phe Glu Val Lys
Ala Phe 1 5 10 15 Ala Thr Asp Ser 20 2520PRTArtificial SequenceHomo
sapiens arrestin epitope 25Gln Val Gln Pro Val Asp Gly Val Val Leu
Val Asp Pro Asp Leu Val 1 5 10 15 Lys Gly Lys Lys 20
2620PRTArtificial SequenceHomo sapiens arrestin epitope 26Arg Val
Gln Val Tyr Pro Pro Val Gly Ala Ala Ser Thr Pro Thr Lys 1 5 10 15
Leu Gln Glu Ser 20 2720PRTArtificial SequenceHomo sapiens arrestin
epitope 27Ser Arg Asp Lys Ser Val Thr Ile Tyr Leu Gly Asn Arg Asp
Tyr Ile 1 5 10 15 Asp His Val Ser 20 2820PRTArtificial SequenceHomo
sapiens arrestin epitope 28Thr Leu Thr Leu Leu Pro Leu Leu Ala Asn
Asn Arg Glu Arg Arg Gly 1 5 10 15 Ile Ala Leu Asp 20
2920PRTArtificial SequenceHomo sapiens arrestin epitope 29Val Ala
Thr Glu Val Pro Phe Arg Leu Met His Pro Gln Pro Glu Asp 1 5 10 15
Pro Ala Lys Glu 20 3020PRTArtificial SequenceHomo sapiens arrestin
epitope 30Val Asp Pro Asp Leu Val Lys Gly Lys Lys Val Tyr Val Thr
Leu Thr 1 5 10 15 Cys Ala Phe Arg 20 3120PRTArtificial SequenceHomo
sapiens arrestin epitope 31Val Val Leu Tyr Ser Ser Asp Tyr Tyr Val
Lys Pro Val Ala Met Glu 1 5 10 15 Glu Ala Gln Glu 20
3220PRTArtificial SequenceHomo sapiens arrestin epitope 32Tyr Gln
Ile Lys Val Lys Leu Thr Val Ser Gly Phe Leu Gly Glu Leu 1 5 10 15
Thr Ser Ser Glu 20 339PRTArtificial SequenceHomo sapiens Chain B,
Structure Of Insulin epitope 33Ala Leu Tyr Leu Val Cys Gly Glu Arg
1 5 3410PRTArtificial SequenceHomo sapiens Chain B, Structure Of
Insulin epitope 34Ser His Leu Val Glu Ala Leu Tyr Leu Val 1 5 10
359PRTArtificial SequenceHomo sapiens chaperonin (HSP60) epitope
35Gln Met Arg Pro Val Ser Arg Val Leu 1 5 369PRTArtificial
SequenceHomo sapiens Collagen alpha-3(IV) chain epitope 36Gly Ser
Pro Ala Thr Trp Thr Thr Arg 1 5 379PRTArtificial SequenceHomo
sapiens collagen, type II, alpha 1 isoform 1 precursor epitope
37Ala Arg Gly Gln Pro Gly Val Met Gly 1 5 389PRTArtificial
SequenceHomo sapiens DNA topoisomerase 1 epitope 38Lys Met Leu Asp
His Glu Tyr Thr Thr 1 5 399PRTArtificial SequenceHomo sapiens ezrin
epitope 39Glu Tyr Thr Ala Lys Ile Ala Leu Leu 1 5 4010PRTArtificial
SequenceHomo sapiens ezrin epitope 40Leu Asn Ile Tyr Glu Lys Asp
Asp Lys Leu 1 5 10 419PRTArtificial SequenceHomo sapiens glial
fibrillary acidic protein isoform 2 epitope 41Asn Leu Ala Gln Asp
Leu Ala Thr Val 1 5 429PRTArtificial SequenceHomo sapiens glial
fibrillary acidic protein isoform 2 epitope 42Gln Leu Ala Arg Gln
Gln Val His Val 1 5 439PRTArtificial SequenceHomo sapiens glucagon
receptor epitope 43Arg Arg Arg Trp His Arg Trp Arg Leu 1 5
449PRTArtificial SequenceHomo sapiens glucose-6-phosphatase,
catalytic, related epitope 44Phe Leu Trp Ser Val Phe Trp Leu Ile 1
5 4515PRTArtificial SequenceHomo sapiens Glutamate decarboxylase 1
epitope 45Asn Met Phe Thr Tyr Glu Ile Ala Pro Val Phe Val Leu Met
Glu 1 5 10 15 4613PRTArtificial SequenceHomo sapiens Glutamate
decarboxylase 2 epitope 46Ile Ala Phe Thr Ser Glu His Ser His Phe
Ser Leu Lys 1 5 10 4713PRTArtificial SequenceHomo sapiens Glutamate
decarboxylase 2 epitope 47Asn Phe Phe Arg Met Val Ile Ser Asn Pro
Ala Ala Thr 1 5 10 489PRTArtificial SequenceHomo sapiens Glutamate
decarboxylase 2 epitope 48Phe Leu Gln Asp Val Met Asn Ile Leu 1 5
499PRTArtificial SequenceHomo sapiens Glutamate decarboxylase 2
epitope 49Leu Leu Gln Glu Tyr Asn Trp Glu Leu 1 5 5010PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 50Arg Met
Met Glu Tyr Gly Thr Thr Met Val 1 5 10 5110PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 51Val Met
Asn Ile Leu Leu Gln Tyr Val Val 1 5 10 5211PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 52Ala Phe
Thr Ser Glu His Ser His Phe Ser Leu 1 5 10 5312PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 53Ala Phe
Thr Ser Glu His Ser His Phe Ser Leu Lys 1 5 10 5411PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 54Phe Lys
Met Phe Pro Glu Val Lys Glu Lys Gly 1 5 10 5510PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 55Phe Thr
Ser Glu His Ser His Phe Ser Leu 1 5 10 5615PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 56Met Ile
Ala Arg Phe Lys Met Phe Pro Glu Val Lys Glu Lys Gly 1 5 10 15
579PRTArtificial SequenceHomo sapiens Glutamate decarboxylase 2
epitope 57Arg Phe Lys Met Phe Pro Glu Val Lys 1 5 5810PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 58Arg Phe
Lys Met Phe Pro Glu Val Lys Glu 1 5 10 5911PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 59Arg Phe
Lys Met Phe Pro Glu Val Lys Glu Lys 1 5 10 609PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 60Thr Ser
Glu His Ser His Phe Ser Leu 1 5 619PRTArtificial SequenceHomo
sapiens Glutamate decarboxylase 2 epitope 61Val Met Asn Ile Leu Leu
Gln Tyr Val 1 5 629PRTArtificial SequenceHomo sapiens Glutamate
decarboxylase 2 epitope 62Glu Leu Ala Glu Tyr Leu Tyr Asn Ile 1 5
639PRTArtificial SequenceHomo sapiens Glutamate decarboxylase 2
epitope 63Ile Leu Met His Cys Gln Thr Thr Leu 1 5 6411PRTArtificial
SequenceHomo sapiens heat shock 27kDa protein 1 epitope 64Gln Leu
Ser Ser Gly Val Ser Glu Ile Arg His 1 5 10 659PRTArtificial
SequenceHomo sapiens HLA class I histocompatibility antigen, B-27
alpha chain precursor epitope 65Leu Arg Arg Tyr Leu Glu Asn Gly Lys
1 5 669PRTArtificial SequenceHomo sapiens HLA class I
histocompatibility antigen, B-7 alpha chain precursor epitope 66Val
Met Ala Pro Arg Thr Val Leu Leu 1 5 6714PRTArtificial SequenceHomo
sapiens HLA class I histocompatibility antigen, B-7 alpha chain
precursor epitope 67Ala Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala
Asp Thr 1 5 10 6814PRTArtificial SequenceHomo sapiens HLA-B27
epitope 68Ala Leu Asn Glu Asp Leu Ser Ser Trp Thr Ala Ala Asp Thr 1
5 10 6910PRTArtificial SequenceHomo sapiens HLA-B27 epitope 69Leu
Leu Arg Gly Tyr His Gln Asp Ala Tyr 1 5 10 7015PRTArtificial
SequenceHomo sapiens HLA-B27 epitope 70Arg Val Ala Glu Gln Leu Arg
Ala Tyr Leu Glu Gly Glu Cys Val 1 5 10 15 7113PRTArtificial
SequenceHomo sapiens HLA-B27 epitope 71Trp Asp Arg Glu Thr Gln Ile
Cys Lys Ala Lys Ala Gln 1 5 10 7211PRTArtificial SequenceHomo
sapiens insulin epitope 72Ala Leu Trp Gly Pro Asp Pro Ala Ala Ala
Phe 1 5 10 7310PRTArtificial SequenceHomo sapiens insulin epitope
73Leu Ala Leu Trp Gly Pro Asp Pro Ala Ala 1 5 10 7411PRTArtificial
SequenceHomo sapiens insulin epitope 74Arg Leu Leu Pro Leu Leu Ala
Leu Leu Ala Leu 1 5 10 759PRTArtificial SequenceHomo sapiens
Insulin precursor epitope 75Ala Leu Trp Met Arg Leu Leu Pro Leu 1 5
769PRTArtificial SequenceHomo sapiens Insulin precursor epitope
76His Leu Val Glu Ala Leu Tyr Leu Val 1 5 7710PRTArtificial
SequenceHomo sapiens Insulin precursor epitope 77Ser Leu Gln Lys
Arg Gly Ile Val Glu Gln 1 5 10 789PRTArtificial SequenceHomo
sapiens Insulin precursor epitope 78Ser Leu Gln Pro Leu Ala Leu Glu
Gly 1 5 799PRTArtificial SequenceHomo sapiens Insulin precursor
epitope 79Ser Leu Tyr Gln Leu Glu Asn Tyr Cys 1 5 8010PRTArtificial
SequenceHomo sapiens Insulin precursor epitope 80Val Cys Gly Glu
Arg Gly Phe Phe Tyr Thr 1 5 10 818PRTArtificial SequenceHomo
sapiens Insulin precursor epitope 81Trp Gly Pro Asp Pro Ala Ala Ala
1 5 829PRTArtificial SequenceHomo sapiens Insulin precursor epitope
82Phe Tyr Thr Pro Lys Thr Arg Arg Glu 1 5 838PRTArtificial
SequenceHomo sapiens Insulin precursor epitope 83Gly Glu Arg Gly
Phe Phe Tyr Thr 1 5 849PRTArtificial SequenceHomo sapiens Insulin
precursor epitope 84Glu Arg Gly Phe Phe Tyr Thr Pro Lys 1 5
8510PRTArtificial SequenceHomo sapiens Insulin precursor epitope
85Leu Cys Gly Ser His Leu Val Glu Ala Leu 1 5 10 8610PRTArtificial
SequenceHomo sapiens Insulin precursor epitope 86Leu Val Cys Gly
Glu Arg Gly Phe Phe Tyr 1 5 10 8710PRTArtificial SequenceHomo
sapiens Insulin precursor epitope 87Leu Tyr Leu Val Cys Gly Glu Arg
Gly Phe 1 5 10 889PRTArtificial SequenceHomo sapiens Islet amyloid
polypeptide precursor epitope 88Phe Leu Ile Val Leu Ser Val Ala Leu
1 5 899PRTArtificial SequenceHomo sapiens Islet amyloid polypeptide
precursor epitope 89Lys Leu Gln Val Phe Leu Ile Val Leu 1 5
909PRTArtificial SequenceHomo sapiens islet-specific
glucose-6-phosphatase-related protein epitope 90Phe Leu Trp Ser Val
Phe Met Leu Ile 1 5 919PRTArtificial SequenceHomo sapiens
islet-specific glucose-6-phosphatase-related protein isoform 1
epitope 91Phe Leu Phe Ala Val Gly Phe Tyr Leu 1 5 929PRTArtificial
SequenceHomo sapiens islet-specific glucose-6-phosphatase-related
protein isoform 1 epitope 92Leu Asn Ile Asp Leu Leu Trp Ser Val 1 5
939PRTArtificial SequenceHomo sapiens islet-specific
glucose-6-phosphatase-related protein isoform 1 epitope 93Val Leu
Phe Gly Leu Gly Phe Ala Ile 1 5 949PRTArtificial SequenceHomo
sapiens islet-specific glucose-6-phosphatase-related protein
isoform 1 epitope 94Asn Leu Phe Leu Phe Leu Phe Ala Val 1 5
959PRTArtificial SequenceHomo sapiens islet-specific
glucose-6-phosphatase-related protein isoform 1 epitope 95Tyr Leu
Leu Leu Arg Val Leu Asn Ile 1 5 969PRTArtificial SequenceHomo
sapiens keratin 6C epitope 96Ala Leu Gln Lys Ala Lys Gln Asp Leu 1
5 979PRTArtificial SequenceHomo sapiens keratin 6C epitope 97Asp
Ala Lys Asn Lys Leu Glu Gly Leu 1 5 989PRTArtificial SequenceHomo
sapiens keratin 6C epitope 98Gly Ala Ser Gly Val Gly Ser Gly Leu 1
5 999PRTArtificial SequenceHomo sapiens keratin 6C epitope 99Lys
Ala Lys Gln Asp Leu Ala Arg Leu 1 5 1009PRTArtificial SequenceHomo
sapiens keratin 6C epitope 100Lys Leu Glu Gly Leu Glu Asp Ala Leu 1
5 1019PRTArtificial SequenceHomo sapiens keratin 6C epitope 101Asn
Met Gln Asp Leu Val Glu Asp Leu 1 5 1029PRTArtificial SequenceHomo
sapiens keratin 6C epitope 102Arg Leu Leu Lys Glu Tyr Gln Glu Leu 1
5 1039PRTArtificial SequenceHomo sapiens keratin 6C epitope 103Trp
Tyr Gln Thr Lys Tyr Glu Glu Leu 1 5
10420PRTArtificial SequenceHomo sapiens Keratin, type I
cytoskeletal 17 (Cytokeratin 17) (K17) (CK 17) (Version 2) epitope
104Leu Arg Arg Val Leu Asp Glu Leu Thr Leu Ala Arg Thr Asp Leu Glu
1 5 10 15 Met Gln Ile Glu 20 1059PRTArtificial SequenceHomo sapiens
Keratin, type I cytoskeletal 17 (Cytokeratin 17) (K17) (CK 17)
(Version 2) epitope 105Ala Leu Glu Glu Ala Asn Ala Asp Leu 1 5
1069PRTArtificial SequenceHomo sapiens Keratin, type I cytoskeletal
17 (Cytokeratin 17) (K17) (CK 17) (Version 2) epitope 106Ala Asn
Ala Asp Leu Glu Val Lys Ile 1 5 1079PRTArtificial SequenceHomo
sapiens Keratin, type I cytoskeletal 17 (Cytokeratin 17) (K17) (CK
17) (Version 2) epitope 107Ala Arg Thr Asp Leu Glu Met Gln Ile 1 5
1089PRTArtificial SequenceHomo sapiens Keratin, type I cytoskeletal
17 (Cytokeratin 17) (K17) (CK 17) (Version 2) epitope 108Ala Ser
Tyr Leu Asp Lys Val Arg Ala 1 5 1099PRTArtificial SequenceHomo
sapiens Keratin, type I cytoskeletal 17 (Cytokeratin 17) (K17) (CK
17) (Version 2) epitope 109Asp Val Asn Gly Leu Arg Arg Val Leu 1 5
1109PRTArtificial SequenceHomo sapiens Keratin, type I cytoskeletal
17 (Cytokeratin 17) (K17) (CK 17) (Version 2) epitope 110Gly Leu
Arg Arg Val Leu Asp Glu Leu 1 5 11112PRTArtificial SequenceHomo
sapiens Keratin, type I cytoskeletal 17 (Cytokeratin 17) (K17) (CK
17) (Version 2) epitope 111Ile Ser Ser Val Leu Ala Gly Ala Ser Cys
Pro Ala 1 5 10 1129PRTArtificial SequenceHomo sapiens Keratin, type
I cytoskeletal 17 (Cytokeratin 17) (K17) (CK 17) (Version 2)
epitope 112Leu Asp Lys Val Arg Ala Leu Glu Glu 1 5
1139PRTArtificial SequenceHomo sapiens Keratin, type I cytoskeletal
17 (Cytokeratin 17) (K17) (CK 17) (Version 2) epitope 113Gln Ile
Glu Gly Leu Lys Glu Glu Leu 1 5 11412PRTArtificial SequenceHomo
sapiens Keratin, type I cytoskeletal 17 (Cytokeratin 17) (K17) (CK
17) (Version 2) epitope 114Arg Ala Leu Glu Glu Ala Asn Ala Asp Leu
Glu Val 1 5 10 1159PRTArtificial SequenceHomo sapiens Keratin, type
I cytoskeletal 17 (Cytokeratin 17) (K17) (CK 17) (Version 2)
epitope 115Arg Leu Ala Ser Tyr Leu Asp Lys Val 1 5
1168PRTArtificial SequenceHomo sapiens Keratin, type I cytoskeletal
17 (Cytokeratin 17) (K17) (CK 17) (Version 2) epitope 116Ser Tyr
Leu Asp Lys Val Arg Ala 1 5 11720PRTArtificial SequenceHomo sapiens
Keratin, type I cytoskeletal 17 (Cytokeratin 17) (K17) (CK 17)
(Version 2) epitope 117Ser Tyr Leu Asp Lys Val Arg Ala Leu Glu Glu
Ala Asn Ala Asp Leu 1 5 10 15 Glu Val Lys Ile 20 1189PRTArtificial
SequenceHomo sapiens maspin epitope 118Gly Leu Glu Lys Ile Glu Lys
Gln Leu 1 5 11910PRTArtificial SequenceHomo sapiens maspin epitope
119Met Gly Asn Ile Asp Ser Ile Asn Cys Lys 1 5 10 1209PRTArtificial
SequenceHomo sapiens maspin epitope 120Tyr Ser Leu Lys Leu Ile Lys
Arg Leu 1 5 12120PRTArtificial SequenceHomo sapiens MBP protein
epitope 121Ala Ser Gln Lys Arg Pro Ser Gln Arg His Gly Ser Lys Tyr
Leu Ala 1 5 10 15 Thr Ala Ser Thr 20 12215PRTArtificial
SequenceHomo sapiens MBP protein epitope 122Glu Asn Pro Val Val His
Phe Phe Lys Asn Ile Val Thr Pro Arg 1 5 10 15 1239PRTArtificial
SequenceHomo sapiens MBP protein epitope 123Val Val His Phe Phe Lys
Asn Ile Val 1 5 12419PRTArtificial SequenceHomo sapiens MBP protein
epitope 124Asp Glu Asn Pro Val Val His Phe Phe Lys Asn Ile Val Thr
Pro Arg 1 5 10 15 Thr Pro Pro 12520PRTArtificial SequenceHomo
sapiens MBP protein epitope 125His His Pro Ala Arg Thr Ala His Tyr
Gly Ser Leu Pro Gln Lys Ser 1 5 10 15 His Gly Arg Thr 20
12620PRTArtificial SequenceHomo sapiens MBP protein epitope 126Val
Val His Phe Phe Lys Asn Ile Val Thr Pro Arg Thr Pro Pro Pro 1 5 10
15 Ser Gln Gly Lys 20 12721PRTArtificial SequenceHomo sapiens MBP
protein epitope 127Ala Ser Gln Lys Arg Pro Ser Gln Arg His Gly Ser
Lys Tyr Leu Ala 1 5 10 15 Thr Ala Ser Thr Met 20 12820PRTArtificial
SequenceHomo sapiens MBP protein epitope 128Phe Lys Gly Val Asp Ala
Gln Gly Thr Leu Ser Lys Ile Phe Lys Leu 1 5 10 15 Gly Gly Arg Asp
20 12919PRTArtificial SequenceHomo sapiens MBP protein epitope
129Arg Pro Gly Phe Gly Tyr Gly Gly Arg Ala Ser Asp Tyr Lys Ser Ala
1 5 10 15 His Lys Gly 13038PRTArtificial SequenceHomo sapiens MBP
protein epitope 130Ala Ser Gln Lys Arg Pro Ser Gln Arg His Gly Ser
Lys Tyr Leu Ala 1 5 10 15 Thr Ala Ser Thr Met Asp His Ala Arg His
Gly Phe Leu Pro Arg His 20 25 30 Arg Asp Thr Gly Ile Leu 35
1319PRTArtificial SequenceHomo sapiens MBP protein epitope 131Lys
Tyr Leu Ala Thr Ala Ser Thr Met 1 5 13220PRTArtificial SequenceHomo
sapiens MBP protein epitope 132Gly Leu Ser Leu Ser Arg Phe Ser Trp
Gly Ala Glu Gly Gln Arg Pro 1 5 10 15 Gly Phe Gly Tyr 20
13343PRTArtificial SequenceHomo sapiens MBP protein epitope 133Phe
Gly Gly Asp Arg Gly Ala Pro Lys Arg Gly Ser Gly Lys Asp Ser 1 5 10
15 His His Pro Ala Arg Thr Ala His Tyr Gly Ser Leu Pro Gln Lys Ser
20 25 30 His Gly Arg Thr Gln Asp Glu Asn Pro Val Val 35 40
13440PRTArtificial SequenceHomo sapiens MBP protein epitope 134Gly
Leu Ser Leu Ser Arg Phe Ser Trp Gly Ala Glu Gly Gln Arg Pro 1 5 10
15 Gly Phe Gly Tyr Gly Gly Arg Ala Ser Asp Tyr Lys Ser Ala His Lys
20 25 30 Gly Phe Lys Gly Val Asp Ala Gln 35 40 1359PRTArtificial
SequenceHomo sapiens MHC class I related protein A epitope 135Ala
Ala Ala Ala Ala Ile Phe Val Ile 1 5 1369PRTArtificial SequenceHomo
sapiens Myelin basic protein epitope 136Ser Leu Ser Arg Phe Ser Trp
Gly Ala 1 5 1379PRTArtificial SequenceHomo sapiens Myelin basic
protein epitope 137Asp Tyr Lys Ser Ala His Lys Gly Phe 1 5
13819PRTArtificial SequenceHomo sapiens myelin basic protein
epitope 138Ser Lys Ile Phe Lys Leu Gly Gly Arg Asp Ser Arg Ser Gly
Ser Pro 1 5 10 15 Met Ala Arg 1398PRTArtificial SequenceHomo
sapiens myelin basic protein epitope 139Thr Pro Arg Thr Pro Pro Pro
Gln 1 5 1409PRTArtificial SequenceHomo sapiens myelin proteolipid
protein epitope 140Phe Leu Tyr Gly Ala Leu Leu Leu Ala 1 5
1419PRTArtificial SequenceHomo sapiens myelin proteolipid protein
epitope 141Lys Leu Ile Glu Thr Tyr Phe Ser Lys 1 5
1429PRTArtificial SequenceHomo sapiens Myelin-associated
glycoprotein precursor epitope 142Leu Met Trp Ala Lys Ile Gly Pro
Val 1 5 1439PRTArtificial SequenceHomo sapiens Myelin-associated
glycoprotein precursor epitope 143Ser Leu Leu Leu Glu Leu Glu Glu
Val 1 5 1449PRTArtificial SequenceHomo sapiens Myelin-associated
glycoprotein precursor epitope 144Val Leu Phe Ser Ser Asp Phe Arg
Ile 1 5 1459PRTArtificial SequenceHomo sapiens Myosin heavy chain,
skeletal muscle, adult 2 (Myosin heavy chain IIa) (MyHC-IIa)
epitope 145Glu Phe Gln Lys Met Arg Arg Asp Leu 1 5
1469PRTArtificial SequenceHomo sapiens Myosin heavy chain, skeletal
muscle, adult 2 (Myosin heavy chain IIa) (MyHC-IIa) epitope 146Lys
Met Arg Arg Asp Leu Glu Glu Ala 1 5 14712PRTArtificial SequenceHomo
sapiens peroxiredoxin-2 isoform a epitope 147Glu Val Lys Leu Ser
Asp Tyr Lys Gly Lys Tyr Val 1 5 10 14810PRTArtificial SequenceHomo
sapiens proinsulin precursor epitope 148His Leu Cys Gly Ser His Leu
Val Glu Ala 1 5 10 14910PRTArtificial SequenceHomo sapiens
proinsulin precursor epitope 149Ala Leu Trp Gly Pro Asp Pro Ala Ala
Ala 1 5 10 1509PRTArtificial SequenceHomo sapiens proinsulin
precursor epitope 150Arg Leu Leu Pro Leu Leu Ala Leu Leu 1 5
15110PRTArtificial SequenceHomo sapiens proinsulin precursor
epitope 151Ala Leu Trp Met Arg Leu Leu Pro Leu Leu 1 5 10
15210PRTArtificial SequenceHomo sapiens proinsulin precursor
epitope 152Trp Met Arg Leu Leu Pro Leu Leu Ala Leu 1 5 10
15310PRTArtificial SequenceHomo sapiens proinsulin precursor
epitope 153Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys 1 5 10
15410PRTArtificial SequenceHomo sapiens proinsulin precursor
epitope 154Pro Leu Leu Ala Leu Leu Ala Leu Trp Gly 1 5 10
1559PRTArtificial SequenceHomo sapiens Receptor-type
tyrosine-protein phosphatase-like N precursor epitope 155Leu Leu
Pro Pro Leu Leu Glu His Leu 1 5 1569PRTArtificial SequenceHomo
sapiens Receptor-type tyrosine-protein phosphatase-like N precursor
epitope 156Ser Leu Ala Ala Gly Val Lys Leu Leu 1 5
1579PRTArtificial SequenceHomo sapiens Receptor-type
tyrosine-protein phosphatase-like N precursor epitope 157Ser Leu
Ser Pro Leu Gln Ala Glu Leu 1 5 1589PRTArtificial SequenceHomo
sapiens Receptor-type tyrosine-protein phosphatase-like N precursor
epitope 158Ala Leu Thr Ala Val Ala Glu Glu Val 1 5
15910PRTArtificial SequenceHomo sapiens Receptor-type
tyrosine-protein phosphatase-like N precursor epitope 159Ser Leu
Tyr His Val Tyr Glu Val Asn Leu 1 5 10 1609PRTArtificial
SequenceHomo sapiens Receptor-type tyrosine-protein
phosphatase-like N precursor epitope 160Thr Ile Ala Asp Phe Trp Gln
Met Val 1 5 1619PRTArtificial SequenceHomo sapiens Receptor-type
tyrosine-protein phosphatase-like N precursor epitope 161Val Ile
Val Met Leu Thr Pro Leu Val 1 5 1629PRTArtificial SequenceHomo
sapiens Receptor-type tyrosine-protein phosphatase-like N precursor
epitope 162Met Val Trp Glu Ser Gly Cys Thr Val 1 5
16314PRTArtificial SequenceHomo sapiens S-arrestin epitope 163Phe
Leu Gly Glu Leu Thr Ser Ser Glu Val Ala Thr Glu Val 1 5 10
16420PRTArtificial SequenceHomo sapiens S-arrestin epitope 164Phe
Met Ser Asp Lys Pro Leu His Leu Ala Val Ser Leu Asn Lys Glu 1 5 10
15 Ile Tyr Phe His 20 16515PRTArtificial SequenceHomo sapiens
S-arrestin epitope 165Gly Glu Ala Glu Glu Gly Lys Arg Asp Lys Asn
Asp Val Asp Glu 1 5 10 15 16620PRTArtificial SequenceHomo sapiens
S-arrestin epitope 166Gly Glu Pro Ile Pro Val Thr Val Thr Val Thr
Asn Asn Thr Glu Lys 1 5 10 15 Thr Val Lys Lys 20 16720PRTArtificial
SequenceHomo sapiens S-arrestin epitope 167His Pro Gln Pro Glu Asp
Pro Ala Lys Glu Ser Tyr Gln Asp Ala Asn 1 5 10 15 Leu Val Phe Glu
20 16820PRTArtificial SequenceHomo sapiens S-arrestin epitope
168Ile Lys Ala Phe Val Glu Gln Val Ala Asn Val Val Leu Tyr Ser Ser
1 5 10 15 Asp Tyr Tyr Val 20 16920PRTArtificial SequenceHomo
sapiens S-arrestin epitope 169Lys Ser Ser Val Arg Leu Leu Ile Arg
Lys Val Gln His Ala Pro Leu 1 5 10 15 Glu Met Gly Pro 20
17020PRTArtificial SequenceHomo sapiens S-arrestin epitope 170Gln
Pro Arg Ala Glu Ala Ala Trp Gln Phe Phe Met Ser Asp Lys Pro 1 5 10
15 Leu His Leu Ala 20 17120PRTArtificial SequenceHomo sapiens
S-arrestin epitope 171Ser Tyr Gln Asp Ala Asn Leu Val Phe Glu Glu
Phe Ala Arg His Asn 1 5 10 15 Leu Lys Asp Ala 20 17220PRTArtificial
SequenceHomo sapiens S-arrestin epitope 172Thr Asp Ala Glu Glu Asp
Lys Ile Pro Lys Lys Ser Ser Val Arg Leu 1 5 10 15 Leu Ile Arg Lys
20 17320PRTArtificial SequenceHomo sapiens S-arrestin epitope
173Thr Asn Asn Thr Glu Lys Thr Val Lys Lys Ile Lys Ala Phe Val Glu
1 5 10 15 Gln Val Ala Asn 20 17420PRTArtificial SequenceHomo
sapiens S-arrestin epitope 174Val Gln His Ala Pro Leu Glu Met Gly
Pro Gln Pro Arg Ala Glu Ala 1 5 10 15 Ala Trp Gln Phe 20
17520PRTArtificial SequenceHomo sapiens S-arrestin epitope 175Val
Ser Leu Asn Lys Glu Ile Tyr Phe His Gly Glu Pro Ile Pro Val 1 5 10
15 Thr Val Thr Val 20 17620PRTArtificial SequenceHomo sapiens
S-arrestin epitope 176Val Tyr Val Thr Leu Thr Cys Ala Phe Arg Tyr
Gly Gln Glu Asp Ile 1 5 10 15 Asp Val Ile Gly 20 17720PRTArtificial
SequenceHomo sapiens S-arrestin epitope 177Tyr Gly Gln Glu Asp Ile
Asp Val Ile Gly Leu Thr Phe Arg Arg Asp 1 5 10 15 Leu Tyr Phe Ser
20 17810PRTArtificial SequenceHomo sapiens SSA protein SS-56
epitope 178Tyr Thr Cys Pro Leu Cys Arg Ala Pro Val 1 5 10
1798PRTArtificial SequenceHomo sapiens Steroid 21-hydroxylase
epitope 179Glu Pro Leu Ala Arg Leu Glu Leu 1 5 18020PRTArtificial
SequenceHomo sapiens Steroid 21-hydroxylase epitope 180Glu Pro Leu
Ala Arg Leu Glu Leu Phe Val Val Leu Thr Arg Leu Leu 1 5 10 15 Gln
Ala Phe Thr 20 18120PRTArtificial SequenceHomo sapiens Steroid
21-hydroxylase epitope 181Ile Lys Asp Asp Asn Leu Met Pro Ala Tyr
Tyr Lys Cys Ile Gln Glu 1 5 10 15 Val Leu Lys Thr 20
18220PRTArtificial SequenceHomo sapiens Steroid 21-hydroxylase
epitope 182Ile Arg Asp Ser Met Glu Pro Val Val Glu Gln Leu Thr Gln
Glu Phe 1 5 10 15 Cys Glu Arg Met 20 1838PRTArtificial SequenceHomo
sapiens T-cell receptor V beta chain 13.1 epitope 183Leu Gly Arg
Ala Gly Leu Thr Tyr 1 5 1849PRTArtificial SequenceHomo sapiens
transaldolase 1 epitope 184Leu Leu Phe Ser Phe Ala Gln Ala Val 1 5
1859PRTArtificial SequenceHomo sapiens Vasoactive intestinal
polypeptide receptor 1 precursor epitope 185Arg Arg Lys Trp Arg Arg
Trp His Leu 1 5 1869PRTArtificial SequenceHomo sapiens Vasoactive
intestinal polypeptide receptor 1 precursor epitope 186Arg Arg Lys
Trp Arg Arg Trp His Leu 1 5 18720PRTArtificial SequenceArachis
hypogaea 2S protein 1 epitope 187Ala His Ala Ser Ala Arg Gln Gln
Trp Glu Leu Gln Gly Asp Arg Arg 1 5 10 15 Cys Gln Ser Gln 20
18820PRTArtificial SequenceArachis hypogaea 2S protein 1 epitope
188Ala Lys Leu Thr Ile Leu Val Ala Leu Ala Leu Phe Leu Leu Ala Ala
1 5 10 15 His Ala Ser Ala 20 18919PRTArtificial SequenceArachis
hypogaea 2S protein 1 epitope 189Ala Leu Gln Gln Ile Met Glu Asn
Gln Ser Asp Arg Leu Gln Gly Arg 1 5 10 15 Gln Gln Glu
19020PRTArtificial SequenceArachis hypogaea 2S protein 1 epitope
190Ala Asn Leu Arg Pro Cys Glu Gln His Leu Met Gln Lys Ile Gln Arg
1 5 10 15 Asp Glu Asp Ser 20 19120PRTArtificial SequenceArachis
hypogaea 2S protein 1 epitope 191Cys Asn Glu Leu Asn Glu Phe Glu
Asn Asn Gln Arg Cys Met Cys Glu 1 5 10 15 Ala Leu Gln Gln 20
19216PRTArtificial SequenceHomo sapiens 5-hydroxytryptamine
receptor 2C (5-HT-2C) (Serotonin receptor 2C) (5-HT2C) (5-HTR2C)
(5HT-1C) epitope 192Pro Arg Gly Thr Met Gln Ala Ile Asn Asn Glu Arg
Lys Ala Ser Lys 1 5 10 15 19312PRTArtificial SequenceBos taurus
Allergen Bos d 2 precursor epitope 193Asp Gln Gly Thr Cys Leu Leu
Leu Thr Glu Val Ala 1 5 10
19414PRTArtificial SequenceBos taurus Allergen Bos d 2 precursor
epitope 194Glu Leu Glu Lys Tyr Gln Gln Leu Asn Ser Glu Arg Gly Val
1 5 10 19513PRTArtificial SequenceBos taurus Allergen Bos d 2
precursor epitope 195Gly Glu Arg Ile Thr Lys Met Thr Glu Gly Leu
Ala Lys 1 5 10 19614PRTArtificial SequenceBos taurus Allergen Bos d
2 precursor epitope 196Pro Gly Glu Trp Arg Ile Ile Tyr Ala Ala Ala
Asp Asn Lys 1 5 10 1978PRTArtificial SequenceBos taurus Allergen
Bos d 2 precursor epitope 197Arg Ile Glu Cys Ile Asn Asp Cys 1 5
19812PRTArtificial SequenceBos taurus Allergen Bos d 2 precursor
epitope 198Val Ala Lys Arg Gln Glu Gly Tyr Val Tyr Val Leu 1 5 10
19910PRTArtificial SequenceBos taurus Allergen Bos d 2 precursor
epitope 199Val Ser Glu Asn Met Leu Val Thr Tyr Val 1 5 10
20012PRTArtificial SequenceBos taurus Allergen Bos d 2 precursor
epitope 200Asp Gln Gly Thr Cys Leu Leu Leu Thr Glu Val Ala 1 5 10
20114PRTArtificial SequenceBos taurus Allergen Bos d 2 precursor
epitope 201Glu Leu Glu Lys Tyr Gln Gln Leu Asn Ser Glu Arg Gly Val
1 5 10 20216PRTArtificial SequenceBos taurus Allergen Bos d 2
precursor epitope 202Glu Leu Glu Lys Tyr Gln Gln Leu Asn Ser Glu
Arg Gly Val Pro Asn 1 5 10 15 20313PRTArtificial SequenceBos taurus
Allergen Bos d 2 precursor epitope 203Gly Glu Arg Ile Thr Lys Met
Thr Glu Gly Leu Ala Lys 1 5 10 20414PRTArtificial SequenceBos
taurus Allergen Bos d 2 precursor epitope 204Pro Gly Glu Trp Arg
Ile Ile Tyr Ala Ala Ala Asp Asn Lys 1 5 10 2058PRTArtificial
SequenceBos taurus Allergen Bos d 2 precursor epitope 205Arg Ile
Glu Cys Ile Asn Asp Cys 1 5 20612PRTArtificial SequenceBos taurus
Allergen Bos d 2 precursor epitope 206Val Ala Lys Arg Gln Glu Gly
Tyr Val Tyr Val Leu 1 5 10 20710PRTArtificial SequenceBos taurus
Allergen Bos d 2 precursor epitope 207Val Ser Glu Asn Met Leu Val
Thr Tyr Val 1 5 10 20813PRTArtificial SequenceCryptomeria japonica
Allergen Cry j 2 epitope 208Asp Ile Phe Ala Ser Lys Asn Phe His Leu
Gln Lys Asn 1 5 10 20913PRTArtificial SequenceCryptomeria japonica
Allergen Cry j 2 epitope 209Gly Ile Ile Ala Ala Tyr Gln Asn Pro Ala
Ser Trp Lys 1 5 10 21012PRTArtificial SequenceCryptomeria japonica
Allergen Cry j 2 epitope 210Lys Leu Thr Ser Gly Lys Ile Ala Ser Cys
Leu Asn 1 5 10 21112PRTArtificial SequenceCryptomeria japonica
Allergen Cry j 2 epitope 211Gln Phe Ala Lys Leu Thr Gly Phe Thr Leu
Met Gly 1 5 10 2128PRTArtificial SequenceAspergillus fumigatus
allergen I/a; Asp f I/a epitope 212Ile Asn Gln Gln Leu Asn Pro Lys
1 5 21315PRTArtificial SequenceAspergillus fumigatus allergen I/a;
Asp f I/a epitope 213Ile Asn Gln Gln Leu Asn Pro Lys Thr Asn Lys
Trp Glu Asp Lys 1 5 10 15 21411PRTArtificial SequenceAspergillus
fumigatus allergen I/a; Asp f I/a epitope 214Leu Asn Pro Lys Thr
Asn Lys Trp Glu Asp Lys 1 5 10 2158PRTArtificial
SequenceAspergillus fumigatus allergen I/a; Asp f I/a epitope
215Ile Asn Gln Gln Leu Asn Pro Lys 1 5 21615PRTArtificial
SequenceAspergillus fumigatus allergen I/a; Asp f I/a epitope
216Ile Asn Gln Gln Leu Asn Pro Lys Thr Asn Lys Trp Glu Asp Lys 1 5
10 15 21711PRTArtificial SequenceAspergillus fumigatus allergen
I/a; Asp f I/a epitope 217Leu Asn Pro Lys Thr Asn Lys Trp Glu Asp
Lys 1 5 10 2187PRTArtificial SequenceAspergillus fumigatus allergen
I/a; Asp f I/a epitope 218Thr Asn Lys Trp Glu Asp Lys 1 5
21912PRTArtificial SequenceAspergillus fumigatus allergen I/a; Asp
f I/a epitope 219Leu Asn Pro Lys Thr Asn Lys Trp Glu Asp Lys Arg 1
5 10 22015PRTArtificial SequenceDermatophagoides farinae Allergen
Mag epitope 220Pro Arg Leu Ser Trp His Gln Tyr Thr Lys Arg Asp Ser
Arg Glu 1 5 10 15 22115PRTArtificial SequenceDermatophagoides
farinae Allergen Mag epitope 221Thr Val Asp Leu Ile Ser Pro Val Thr
Lys Arg Ala Ser Leu Lys 1 5 10 15 22218PRTArtificial SequenceBos
taurus Alpha-S1-casein precursor epitope 222Ala Trp Tyr Tyr Val Pro
Leu Gly Thr Gln Tyr Thr Asp Ala Pro Ser 1 5 10 15 Phe Ser
22318PRTArtificial SequenceBos taurus Alpha-S1-casein precursor
epitope 223Asp Ala Tyr Pro Ser Gly Ala Trp Tyr Tyr Val Pro Leu Gly
Thr Gln 1 5 10 15 Tyr Thr 22418PRTArtificial SequenceBos taurus
Alpha-S1-casein precursor epitope 224Asp Ile Gly Ser Glu Ser Thr
Glu Asp Gln Ala Met Glu Asp Ile Lys 1 5 10 15 Gln Met
2256PRTArtificial SequenceBos taurus Alpha-S1-casein precursor
epitope 225Glu Asp Ile Lys Gln Met 1 5 22612PRTArtificial
SequenceBos taurus Alpha-S1-casein precursor epitope 226Glu Pro Met
Ile Gly Val Asn Gln Glu Leu Ala Tyr 1 5 10 22718PRTArtificial
SequenceBos taurus Alpha-S1-casein precursor epitope 227Glu Pro Met
Ile Gly Val Asn Gln Glu Leu Ala Tyr Phe Tyr Pro Glu 1 5 10 15 Leu
Phe 22817PRTArtificial SequenceArachis hypogaea Ara h 2.01 allergen
epitope 228Glu Leu Asn Glu Phe Glu Asn Asn Gln Arg Cys Met Cys Glu
Ala Leu 1 5 10 15 Gln 22916PRTArtificial SequenceArachis hypogaea
Ara h 2.01 allergen epitope 229Ser Gln Leu Glu Arg Ala Asn Leu Arg
Pro Cys Glu Gln His Leu Met 1 5 10 15 23015PRTArtificial
SequenceCryptomeria japonica Cry j 1 precursor epitope 230Gly Ala
Thr Arg Asp Arg Pro Leu Trp Ile Ile Phe Ser Gly Asn 1 5 10 15
23120PRTArtificial SequenceCryptomeria japonica Cry j 1 precursor
epitope 231Ile Phe Ser Gly Asn Met Asn Ile Lys Leu Lys Met Pro Met
Tyr Ile 1 5 10 15 Ala Gly Tyr Lys 20 23220PRTArtificial
SequenceCryptomeria japonica Cry j 1 precursor epitope 232Lys Met
Pro Met Tyr Ile Ala Gly Tyr Lys Thr Phe Asp Gly Arg Gly 1 5 10 15
Ala Gln Val Tyr 20 23320PRTArtificial SequenceCryptomeria japonica
Cry j 1 precursor epitope 233Leu Gly His Asp Asp Ala Tyr Ser Asp
Asp Lys Ser Met Lys Val Thr 1 5 10 15 Val Ala Phe Asn 20
23419PRTArtificial SequenceCryptomeria japonica Cry j 1 precursor
epitope 234Ser Gly Lys Tyr Glu Gly Gly Asn Ile Tyr Thr Lys Lys Glu
Ala Phe 1 5 10 15 Asn Val Glu 23511PRTArtificial
SequenceCochliobolus lunatus Cytochrome c epitope 235Glu Asn Pro
Lys Lys Tyr Ile Pro Gly Thr Lys 1 5 10 23611PRTArtificial
SequenceCochliobolus lunatus Cytochrome c epitope 236Gly Leu Phe
Gly Arg Lys Thr Gly Ser Val Ala 1 5 10 2379PRTArtificial
SequenceCochliobolus lunatus Cytochrome c epitope 237Lys Ile Gly
Pro Glu Leu His Gly Leu 1 5 23812PRTArtificial SequenceCochliobolus
lunatus Cytochrome c epitope 238Leu Lys Ala Gly Glu Gly Asn Lys Ile
Gly Pro Glu 1 5 10 23911PRTArtificial SequenceCochliobolus lunatus
Cytochrome c epitope 239Leu Lys Lys Pro Lys Asp Arg Asn Asp Leu Ile
1 5 10 24018PRTArtificial SequenceDermatophagoides farinae Der f 2
allergen epitope 240Gly Leu Glu Ile Asp Val Pro Gly Ile Asp Thr Asn
Ala Cys His Phe 1 5 10 15 Val Lys 24120PRTArtificial
SequenceDermatophagoides farinae Der f 2 allergen epitope 241Pro
Gly Ile Asp Thr Asn Ala Cys His Phe Val Lys Cys Pro Leu Val 1 5 10
15 Lys Gly Gln Gln 20 24219PRTArtificial SequenceDermatophagoides
pteronyssinus Der p 1 allergen epitope 242Arg Phe Gly Ile Ser Asn
Tyr Cys Gln Ile Tyr Pro Pro Asn Ala Asn 1 5 10 15 Lys Ile Arg
24315PRTArtificial SequenceDermatophagoides pteronyssinus Der p 1
allergen epitope 243Ala Val Asn Ile Val Gly Tyr Ser Asn Ala Gln Gly
Val Asp Tyr 1 5 10 15 24420PRTArtificial SequenceChironomus thummi
globin Ctt 3-1 epitope 244Phe Ala Gly Lys Asp Leu Glu Ser Ile Lys
Gly Thr Ala Pro Phe Glu 1 5 10 15 Thr His Ala Asn 20
24511PRTArtificial SequenceChironomus thummi globin Ctt 3-1 epitope
245Gly Thr Ala Pro Phe Glu Thr His Ala Asn Arg 1 5 10
24621PRTArtificial SequenceChironomus thummi globin Ctt 3-1 epitope
246Lys Gly Thr Ala Pro Phe Glu Thr His Ala Asn Arg Ile Val Gly Phe
1 5 10 15 Phe Ser Lys Ile Ile 20 24721PRTArtificial
SequenceChironomus thummi thummi Globin CTT-III epitope 247Ala His
Thr Asp Phe Ala Gly Ala Glu Ala Ala Trp Gly Ala Thr Leu 1 5 10 15
Asp Thr Phe Phe Gly 20 24820PRTArtificial SequenceChironomus thummi
thummi Globin CTT-III epitope 248Phe Ala Gly Lys Asp Leu Glu Ser
Ile Lys Gly Thr Ala Pro Phe Glu 1 5 10 15 Ile His Ala Asn 20
24921PRTArtificial SequenceChironomus thummi thummi Globin CTT-III
epitope 249Val Asn Thr Phe Val Ala Ser His Lys Pro Arg Gly Val Thr
His Asp 1 5 10 15 Gln Leu Asn Asn Phe 20 2508PRTArtificial
SequenceChironomus thummi thummi Globin CTT-III precursor epitope
250Ala Asp Pro Ser Ile Met Ala Lys 1 5 25121PRTArtificial
SequenceChironomus thummi thummi Globin CTT-III precursor epitope
251Ala Asp Pro Ser Ile Met Ala Lys Phe Thr Gln Phe Ala Gly Lys Asp
1 5 10 15 Leu Glu Ser Ile Lys 20 2525PRTArtificial
SequenceChironomus thummi thummi Globin CTT-III precursor epitope
252Ala Glu Ala Ala Trp 1 5 25320PRTArtificial SequenceChironomus
thummi thummi Globin CTT-III precursor epitope 253Ala Glu Ala Ala
Trp Gly Ala Thr Leu Asp Thr Phe Phe Gly Met Ile 1 5 10 15 Phe Ser
Lys Met 20 2548PRTArtificial SequenceChironomus thummi thummi
Globin CTT-III precursor epitope 254Ala Gly Phe Val Ser Tyr Met Lys
1 5 25515PRTArtificial SequencePhaseolus vulgaris Glycine-rich cell
wall structural protein 1.8 precursor epitope 255Gly Gly Tyr Gly
Asp Gly Gly Ala His Gly Gly Gly Tyr Gly Gly 1 5 10 15
25615PRTArtificial SequencePhleum pratense Group V allergen Phl p 5
epitope 256Ala Thr Pro Glu Ala Lys Tyr Asp Ala Tyr Val Ala Thr Leu
Ser 1 5 10 15 25715PRTArtificial SequencePhleum pratense Group V
allergen Phl p 5 epitope 257Phe Thr Val Phe Glu Ala Ala Phe Asn Asn
Ala Ile Lys Ala Gly 1 5 10 15 25815PRTArtificial SequencePhleum
pratense Group V allergen Phl p 5 epitope 258Lys Tyr Asp Ala Tyr
Val Ala Thr Leu Ser Glu Ala Leu Arg Ile 1 5 10 15
25915PRTArtificial SequencePhleum pratense Group V allergen Phl p 5
epitope 259Pro Ala Asn Asp Lys Phe Thr Val Phe Glu Ala Ala Phe Asn
Asn 1 5 10 15 26015PRTArtificial SequencePhleum pratense Group V
allergen Phl p 5 epitope 260Pro Lys Gly Gly Ala Glu Ser Ser Ser Lys
Ala Ala Leu Thr Ser 1 5 10 15 26116PRTArtificial SequenceHomo
sapiens KIAA1224 protein epitope 261Asp Leu Glu Ser Tyr Leu Gln Leu
Asn Cys Glu Arg Gly Thr Trp Arg 1 5 10 15 26215PRTArtificial
SequenceLepidoglyphus destructor Lep D 2 precursor epitope 262Lys
Gly Glu Ala Leu Asp Phe Asn Tyr Gly Met Thr Ile Pro Ala 1 5 10 15
26312PRTArtificial SequenceCorylus avellana lipid transfer protein
precursor epitope 263Ala Gly Leu Pro Gly Lys Cys Gly Val Asn Ile
Pro 1 5 10 26412PRTArtificial SequenceCorylus avellana lipid
transfer protein precursor epitope 264Ala Lys Gly Ile Ala Gly Leu
Asn Pro Asn Leu Ala 1 5 10 26512PRTArtificial SequenceCorylus
avellana lipid transfer protein precursor epitope 265Cys Gly Val
Asn Ile Pro Tyr Lys Ile Ser Pro Ser 1 5 10 26612PRTArtificial
SequenceCorylus avellana lipid transfer protein precursor epitope
266Cys Lys Gly Val Arg Ala Val Asn Asp Ala Ser Arg 1 5 10
26712PRTArtificial SequenceCorylus avellana lipid transfer protein
precursor epitope 267Cys Val Leu Tyr Leu Lys Asn Gly Gly Val Leu
Pro 1 5 10 26816PRTArtificial SequenceHomo sapiens Lipocalin 1
(tear prealbumin) epitope 268Lys Pro Val Arg Gly Val Lys Leu Val
Gly Arg Asp Pro Lys Asn Asn 1 5 10 15 26915PRTArtificial
SequenceDermatophagoides farinae Mag3 epitope 269Glu Phe Asn Thr
Glu Phe Thr Ile His Ala Asp Lys Asn Asn Leu 1 5 10 15
27015PRTArtificial SequenceDermatophagoides farinae Mag3 epitope
270Phe Thr Ile His Ala Asp Lys Asn Asn Leu Lys Met His Met Asp 1 5
10 15 27115PRTArtificial SequenceDermatophagoides farinae Mag3
epitope 271Lys Met His Met Asp Phe Pro Asn Val Phe Gln Ala Asp Leu
Thr 1 5 10 15 27213PRTArtificial SequenceApium graveolens Major
allergen Api g 1 epitope 272Ala Leu Phe Lys Ala Leu Glu Ala Tyr Leu
Ile Ala Asn 1 5 10 27312PRTArtificial SequenceApium graveolens
Major allergen Api g 1 epitope 273Asp Ala Val Val Pro Glu Glu Asn
Ile Lys Tyr Ala 1 5 10 27412PRTArtificial SequenceApium graveolens
Major allergen Api g 1 epitope 274Asp Ile Leu Leu Gly Phe Ile Glu
Ser Ile Glu Asn 1 5 10 27512PRTArtificial SequenceApium graveolens
Major allergen Api g 1 epitope 275Gly Gly Ser Ile Cys Lys Thr Thr
Ala Ile Phe His 1 5 10 27612PRTArtificial SequenceApium graveolens
Major allergen Api g 1 epitope 276Gly Val Gln Thr His Val Leu Glu
Leu Thr Ser Ser 1 5 10 27715PRTArtificial SequenceAspergillus
fumigatus Major allergen Asp f 2 precursor epitope 277Phe Gly Asn
Arg Pro Thr Met Glu Ala Val Gly Ala Tyr Asp Val 1 5 10 15
27815PRTArtificial SequenceAspergillus fumigatus Major allergen Asp
f 2 precursor epitope 278Met Glu Ala Val Gly Ala Tyr Asp Val Ile
Val Asn Gly Asp Lys 1 5 10 15 27916PRTArtificial SequenceCanis
lupus familiaris Major allergen Can f 1 precursor epitope 279Ala
Leu Glu Asp Phe Arg Glu Phe Ser Arg Ala Lys Gly Leu Asn Gln 1 5 10
15 28016PRTArtificial SequenceCanis lupus familiaris Major allergen
Can f 1 precursor epitope 280Asp Gln Glu Val Pro Glu Lys Pro Asp
Ser Val Thr Pro Met Ile Leu 1 5 10 15 28112PRTArtificial
SequenceCorylus avellana major allergen Cor a 1.0401 epitope 281Ala
Gly Lys Glu Lys Ala Ala Gly Leu Phe Lys Ala 1 5 10
28212PRTArtificial SequenceCorylus avellana major allergen Cor a
1.0401 epitope 282Ala Gly Leu Phe Lys Ala Val Glu Ala Tyr Leu Leu 1
5 10 28312PRTArtificial SequenceCorylus avellana major allergen Cor
a 1.0401 epitope 283Ala Pro Gln His Phe Thr Ser Ala Glu Asn Leu Glu
1 5 10 28412PRTArtificial SequenceCorylus avellana major allergen
Cor a 1.0401 epitope 284Ala Arg Leu Phe Lys Ser Phe Val Leu Asp Ala
Asp 1 5 10 28512PRTArtificial SequenceCorylus avellana major
allergen Cor a 1.0401 epitope 285Glu Ile Asp His Ala Asn Phe Lys
Tyr Cys Tyr Ser 1 5 10 28613PRTArtificial SequenceDaucus carota
Major allergen Dau c 1 epitope 286Ala Leu Phe Lys Ala Ile Glu Ala
Tyr Leu Ile Ala Asn 1 5 10 28716PRTArtificial SequenceEquus
caballus Major allergen Equ c 1
precursor epitope 287Asp Gly Tyr Asn Val Phe Arg Ile Ser Glu Phe
Glu Asn Asp Glu His 1 5 10 15 28816PRTArtificial SequenceEquus
caballus Major allergen Equ c 1 precursor epitope 288Asp Lys Asp
Arg Pro Phe Gln Leu Phe Glu Phe Tyr Ala Arg Glu Pro 1 5 10 15
28916PRTArtificial SequenceEquus caballus Major allergen Equ c 1
precursor epitope 289Asp Leu Thr Lys Ile Asp Arg Cys Phe Gln Leu
Arg Gly Asn Gly Val 1 5 10 15 29016PRTArtificial SequenceEquus
caballus Major allergen Equ c 1 precursor epitope 290Asp Arg Pro
Phe Gln Leu Phe Glu Phe Tyr Ala Arg Glu Pro Asp Val 1 5 10 15
29116PRTArtificial SequenceEquus caballus Major allergen Equ c 1
precursor epitope 291Asp Val Ser Pro Glu Ile Lys Glu Glu Phe Val
Lys Ile Val Gln Lys 1 5 10 15 29217PRTArtificial SequenceFelis
catus major allergen I epitope 292Glu Asn Ala Arg Ile Leu Lys Asn
Cys Val Asp Ala Lys Met Thr Glu 1 5 10 15 Glu 29317PRTArtificial
SequenceFelis catus major allergen I epitope 293Arg Asp Val Asp Leu
Phe Leu Thr Gly Thr Pro Asp Glu Tyr Val Glu 1 5 10 15 Gln
29417PRTArtificial SequenceFelis catus major allergen I epitope
294Thr Gly Thr Pro Asp Glu Tyr Val Glu Gln Val Ala Gln Tyr Lys Ala
1 5 10 15 Leu 29517PRTArtificial SequenceFelis catus Major allergen
I polypeptide chain 1 precursor epitope 295Asp Val Asp Leu Phe Leu
Thr Gly Thr Pro Asp Glu Tyr Val Glu Gln 1 5 10 15 Val
29617PRTArtificial SequenceFelis catus Major allergen I polypeptide
chain 1 precursor epitope 296Glu Ile Cys Pro Ala Val Lys Arg Asp
Val Asp Leu Phe Leu Thr Gly 1 5 10 15 Thr 29716PRTArtificial
SequenceFelis catus Major allergen I polypeptide chain 1 precursor
epitope 297Glu Gln Val Ala Gln Tyr Lys Ala Leu Pro Val Val Leu Glu
Asn Ala 1 5 10 15 29817PRTArtificial SequenceFelis catus Major
allergen I polypeptide chain 1 precursor epitope 298Lys Ala Leu Pro
Val Val Leu Glu Asn Ala Arg Ile Leu Lys Asn Cys 1 5 10 15 Val
29917PRTArtificial SequenceFelis catus Major allergen I polypeptide
chain 1 precursor epitope 299Leu Phe Leu Thr Gly Thr Pro Asp Glu
Tyr Val Glu Gln Val Ala Gln 1 5 10 15 Tyr 30016PRTArtificial
SequenceFelis catus major allergen I, polypeptide chain 1 epitope
300Lys Glu Asn Ala Leu Ser Leu Leu Asp Lys Ile Tyr Thr Ser Pro Leu
1 5 10 15 30116PRTArtificial SequenceFelis catus major allergen I,
polypeptide chain 1 epitope 301Lys Met Thr Glu Glu Asp Lys Glu Asn
Ala Leu Ser Leu Leu Asp Lys 1 5 10 15 30215PRTArtificial
SequenceMalus x domestica Major allergen Mal d 1 epitope 302Gly Leu
Phe Lys Leu Ile Glu Ser Tyr Leu Lys Asp His Pro Asp 1 5 10 15
30315PRTArtificial SequencePrunus avium Major allergen Pru av 1
epitope 303Asn Leu Phe Lys Leu Ile Glu Thr Tyr Leu Lys Gly His Pro
Asp 1 5 10 15 30420PRTArtificial SequenceHevea brasiliensis Major
latex allergen Hev b 5 epitope 304Ala Ala Pro Ala Glu Gly Glu Lys
Pro Ala Glu Glu Glu Lys Pro Ile 1 5 10 15 Thr Glu Ala Ala 20
30520PRTArtificial SequenceHevea brasiliensis Major latex allergen
Hev b 5 epitope 305Ala Glu Glu Glu Lys Pro Ile Thr Glu Ala Ala Glu
Thr Ala Thr Thr 1 5 10 15 Glu Val Pro Val 20 30620PRTArtificial
SequenceHevea brasiliensis Major latex allergen Hev b 5 epitope
306Ala Pro Ala Glu Pro Glu Ala Pro Ala Pro Glu Thr Glu Lys Ala Glu
1 5 10 15 Glu Val Glu Lys 20 30720PRTArtificial SequenceHevea
brasiliensis Major latex allergen Hev b 5 epitope 307Ala Pro Glu
Ala Asp Gln Thr Thr Pro Glu Glu Lys Pro Ala Glu Pro 1 5 10 15 Glu
Pro Val Ala 20 30820PRTArtificial SequenceHevea brasiliensis Major
latex allergen Hev b 5 epitope 308Ala Ser Glu Gln Glu Thr Ala Asp
Ala Thr Pro Glu Lys Glu Glu Pro 1 5 10 15 Thr Ala Ala Pro 20
30911PRTArtificial SequenceDermatophagoides pteronyssinus Major
mite fecal allergen Der p 1 epitope 309Tyr Ala Tyr Val Ala Arg Glu
Gln Ser Cys Arg 1 5 10 31019PRTArtificial SequenceDermatophagoides
pteronyssinus Major mite fecal allergen Der p 1 epitope 310Ala Leu
Ala Gln Thr His Thr Ala Ile Ala Val Ile Ile Gly Ile Lys 1 5 10 15
Asp Leu Asp 31135PRTArtificial SequenceOlea europaea Major pollen
allergen epitope 311Glu Asp Ile Pro Gln Pro Pro Val Ser Gln Phe His
Ile Gln Gly Gln 1 5 10 15 Val Tyr Cys Asp Thr Cys Arg Ala Gly Phe
Ile Thr Glu Leu Ser Glu 20 25 30 Phe Ile Pro 35 31231PRTArtificial
SequenceOlea europaea Major pollen allergen epitope 312Gly Ala Ser
Leu Arg Leu Gln Cys Lys Asp Lys Glu Asn Gly Asp Val 1 5 10 15 Thr
Phe Thr Glu Val Gly Tyr Thr Arg Ala Glu Gly Leu Tyr Ser 20 25 30
31334PRTArtificial SequenceOlea europaea Major pollen allergen
epitope 313Gly Thr Thr Arg Thr Val Asn Pro Leu Gly Phe Phe Lys Lys
Glu Ala 1 5 10 15 Leu Pro Lys Cys Ala Gln Val Tyr Asn Lys Leu Gly
Met Tyr Pro Pro 20 25 30 Asn Met 31453PRTArtificial SequenceOlea
europaea Major pollen allergen epitope 314Leu Val Glu Arg Asp His
Lys Asn Glu Phe Cys Glu Ile Thr Leu Ile 1 5 10 15 Ser Ser Gly Arg
Lys Asp Cys Asn Glu Ile Pro Thr Glu Gly Trp Ala 20 25 30 Lys Pro
Ser Leu Lys Phe Lys Leu Asn Thr Val Asn Gly Thr Thr Arg 35 40 45
Thr Val Asn Pro Leu 50 31533PRTArtificial SequenceOlea europaea
Major pollen allergen epitope 315Met Leu Val Glu Arg Asp His Lys
Asn Glu Phe Cys Glu Ile Thr Leu 1 5 10 15 Ile Ser Ser Gly Arg Lys
Asp Cys Asn Glu Ile Pro Thr Glu Gly Trp 20 25 30 Ala
31612PRTArtificial SequenceArtemisia vulgaris Major pollen allergen
Art v 1 precursor epitope 316Ala Gly Gly Ser Pro Ser Pro Pro Ala
Asp Gly Gly 1 5 10 31712PRTArtificial SequenceArtemisia vulgaris
Major pollen allergen Art v 1 precursor epitope 317Ala Gly Ser Lys
Leu Cys Glu Lys Thr Ser Lys Thr 1 5 10 31812PRTArtificial
SequenceArtemisia vulgaris Major pollen allergen Art v 1 precursor
epitope 318Cys Asp Lys Lys Cys Ile Glu Trp Glu Lys Ala Gln 1 5 10
31912PRTArtificial SequenceArtemisia vulgaris Major pollen allergen
Art v 1 precursor epitope 319Asp Gly Gly Ser Pro Pro Pro Pro Ala
Asp Gly Gly 1 5 10 32012PRTArtificial SequenceArtemisia vulgaris
Major pollen allergen Art v 1 precursor epitope 320Glu Lys Thr Ser
Lys Thr Tyr Ser Gly Lys Cys Asp 1 5 10 32112PRTArtificial
SequenceBetula pendula Major pollen allergen Bet v 1-A epitope
321Ala Ala Arg Leu Phe Lys Ala Phe Ile Leu Asp Gly 1 5 10
32215PRTArtificial SequenceBetula pendula Major pollen allergen Bet
v 1-A epitope 322Ala Ala Arg Leu Phe Lys Ala Phe Ile Leu Asp Gly
Asp Asn Leu 1 5 10 15 32312PRTArtificial SequenceBetula pendula
Major pollen allergen Bet v 1-A epitope 323Ala Glu Gln Val Lys Ala
Ser Lys Glu Met Gly Glu 1 5 10 32421PRTArtificial SequenceBetula
pendula Major pollen allergen Bet v 1-A epitope 324Ala Phe Ile Leu
Asp Gly Asp Asn Leu Phe Pro Lys Val Ala Pro Gln 1 5 10 15 Ala Ile
Ser Ser Val 20 32512PRTArtificial SequenceBetula pendula Major
pollen allergen Bet v 1-A epitope 325Ala Ile Ser Ser Val Glu Asn
Ile Glu Gly Asn Gly 1 5 10 32615PRTArtificial SequenceBetula
pendula Major pollen allergen Bet v 1-A epitope 326Glu Thr Leu Leu
Arg Ala Val Glu Ser Tyr Leu Leu Ala His Ser 1 5 10 15
32716PRTArtificial SequenceBetula pendula Major pollen allergen Bet
v 1-F/I epitope 327Gly Glu Thr Leu Leu Arg Ala Val Glu Ser Tyr Leu
Leu Ala His Ser 1 5 10 15 32820PRTArtificial SequenceChamaecyparis
obtusa Major pollen allergen Cha o 1 precursor epitope 328Ala Asn
Asn Asn Tyr Asp Pro Trp Ser Ile Tyr Ala Ile Gly Gly Ser 1 5 10 15
Ser Asn Pro Thr 20 32920PRTArtificial SequenceChamaecyparis obtusa
Major pollen allergen Cha o 1 precursor epitope 329Ala Ser Thr Gly
Val Thr Ile Ser Asn Asn His Phe Phe Asn His His 1 5 10 15 Lys Val
Met Leu 20 33020PRTArtificial SequenceChamaecyparis obtusa Major
pollen allergen Cha o 1 precursor epitope 330Cys Ala Asn Trp Val
Trp Arg Ser Thr Gln Asp Ser Phe Asn Asn Gly 1 5 10 15 Ala Tyr Phe
Val 20 33120PRTArtificial SequenceChamaecyparis obtusa Major pollen
allergen Cha o 1 precursor epitope 331Asp Ala Ile Thr Met Arg Asn
Val Thr Asp Val Trp Ile Asp His Asn 1 5 10 15 Ser Leu Ser Asp 20
33220PRTArtificial SequenceChamaecyparis obtusa Major pollen
allergen Cha o 1 precursor epitope 332Asp Ala Asn Trp Asp Gln Asn
Arg Met Lys Leu Ala Asp Cys Ala Val 1 5 10 15 Gly Phe Gly Ser 20
33320PRTArtificial SequenceCynodon dactylon Major pollen allergen
Cyn d 1 epitope 333Ala Ile Gly Asp Lys Pro Gly Pro Asn Ile Thr Ala
Thr Tyr Gly Asn 1 5 10 15 Lys Trp Leu Glu 20 33420PRTArtificial
SequenceCynodon dactylon Major pollen allergen Cyn d 1 epitope
334Cys Tyr Glu Ile Lys Cys Lys Glu Pro Val Glu Cys Ser Gly Glu Pro
1 5 10 15 Val Leu Val Lys 20 33520PRTArtificial SequenceCynodon
dactylon Major pollen allergen Cyn d 1 epitope 335Asp His Gly Gly
Ala Cys Gly Tyr Lys Asp Val Asp Lys Pro Pro Phe 1 5 10 15 Asp Gly
Met Thr 20 33620PRTArtificial SequenceCynodon dactylon Major pollen
allergen Cyn d 1 epitope 336Glu Gly Gly Ala His Leu Val Gln Asp Asp
Val Ile Pro Ala Asn Trp 1 5 10 15 Lys Pro Asp Thr 20
33720PRTArtificial SequenceCynodon dactylon Major pollen allergen
Cyn d 1 epitope 337Phe Lys Asp Gly Leu Gly Cys Gly Ala Cys Tyr Glu
Ile Lys Cys Lys 1 5 10 15 Glu Pro Val Glu 20 33815PRTArtificial
SequencePhleum pratense Major pollen allergen Phl p 4 precursor
epitope 338Phe Ala Glu Tyr Lys Ser Asp Tyr Val Tyr Gln Pro Phe Pro
Lys 1 5 10 15 33915PRTArtificial SequencePhleum pratense Major
pollen allergen Phl p 4 precursor epitope 339Met Leu Leu Arg Lys
Tyr Gly Ile Ala Ala Glu Asn Val Ile Asp 1 5 10 15
34015PRTArtificial SequencePhleum pratense Major pollen allergen
Phl p 4 precursor epitope 340Asn Ser Phe Lys Pro Phe Ala Glu Tyr
Lys Ser Asp Tyr Val Tyr 1 5 10 15 34120PRTArtificial SequenceRattus
norvegicus Major urinary protein precursor epitope 341Ala Ser Asn
Lys Arg Glu Lys Ile Glu Glu Asn Gly Ser Met Arg Val 1 5 10 15 Phe
Met Gln His 20 34220PRTArtificial SequenceRattus norvegicus Major
urinary protein precursor epitope 342Asp Ile Lys Glu Lys Phe Ala
Lys Leu Cys Glu Ala His Gly Ile Thr 1 5 10 15 Arg Asp Asn Ile 20
34320PRTArtificial SequenceRattus norvegicus Major urinary protein
precursor epitope 343Glu Glu Ala Ser Ser Thr Arg Gly Asn Leu Asp
Val Ala Lys Leu Asn 1 5 10 15 Gly Asp Trp Phe 20 34420PRTArtificial
SequenceRattus norvegicus Major urinary protein precursor epitope
344Glu Glu Asn Gly Ser Met Arg Val Phe Met Gln His Ile Asp Val Leu
1 5 10 15 Glu Asn Ser Leu 20 34520PRTArtificial SequenceRattus
norvegicus Major urinary protein precursor epitope 345Glu Asn Ser
Leu Gly Phe Lys Phe Arg Ile Lys Glu Asn Gly Glu Cys 1 5 10 15 Arg
Glu Leu Tyr 20 34621PRTArtificial SequenceDermatophagoides farinae
Mite group 2 allergen Der f 2 precursor epitope 346Asp Ile Lys Tyr
Thr Trp Asn Val Pro Lys Ile Ala Pro Lys Ser Glu 1 5 10 15 Asn Val
Val Val Thr 20 34717PRTArtificial SequenceDermatophagoides farinae
Mite group 2 allergen Der f 2 precursor epitope 347Asp Asn Gly Val
Leu Ala Cys Ala Ile Ala Thr His Gly Lys Ile Arg 1 5 10 15 Asp
34821PRTArtificial SequenceDermatophagoides farinae Mite group 2
allergen Der f 2 precursor epitope 348Glu Ala Leu Phe Asp Ala Asn
Gln Asn Thr Lys Thr Ala Lys Ile Glu 1 5 10 15 Ile Lys Ala Ser Leu
20 34945PRTArtificial SequenceDermatophagoides farinae Mite group 2
allergen Der f 2 precursor epitope 349Gln Tyr Asp Ile Lys Tyr Thr
Trp Asn Val Pro Lys Ile Ala Pro Lys 1 5 10 15 Ser Glu Asn Val Val
Val Thr Val Lys Leu Ile Gly Asp Asn Gly Val 20 25 30 Leu Ala Cys
Ala Ile Ala Thr His Gly Lys Ile Arg Asp 35 40 45 35019PRTArtificial
SequenceDermatophagoides farinae Mite group 2 allergen Der f 2
precursor epitope 350Thr Lys Thr Ala Lys Ile Glu Ile Lys Ala Ser
Leu Asp Gly Leu Glu 1 5 10 15 Ile Asp Val 35114PRTArtificial
SequenceDermatophagoides pteronyssinus Mite group 2 allergen Der p
2 epitope 351Ala Ser Ile Asp Gly Leu Gly Val Asp Val Pro Gly Ile
Asp 1 5 10 35215PRTArtificial SequenceDermatophagoides
pteronyssinus Mite group 2 allergen Der p 2 epitope 352Phe Glu Ala
Val Gln Asn Thr Lys Thr Ala Lys Ile Glu Ile Lys 1 5 10 15
35317PRTArtificial SequenceDermatophagoides pteronyssinus Mite
group 2 allergen Der p 2 epitope 353Arg Gly Lys Pro Pro Gln Leu Glu
Ala Val Phe Glu Ala Val Gln Asn 1 5 10 15 Thr 35415PRTArtificial
SequenceDermatophagoides pteronyssinus Mite group 2 allergen Der p
2 precursor epitope 354Cys His Gly Ser Glu Pro Cys Ile Ile His Arg
Gly Lys Pro Phe 1 5 10 15 35527PRTArtificial
SequenceDermatophagoides pteronyssinus Mite group 2 allergen Der p
2 precursor epitope 355Cys Pro Leu Val Lys Gly Gln Gln Tyr Asp Ile
Lys Tyr Thr Trp Asn 1 5 10 15 Val Pro Lys Ile Ala Pro Lys Ser Glu
Asn Val 20 25 35626PRTArtificial SequenceDermatophagoides
pteronyssinus Mite group 2 allergen Der p 2 precursor epitope
356Asp Ile Lys Tyr Thr Trp Asn Val Pro Lys Ile Ala Pro Lys Ser Glu
1 5 10 15 Asn Val Val Val Thr Val Lys Val Met Gly 20 25
35715PRTArtificial SequenceDermatophagoides pteronyssinus Mite
group 2 allergen Der p 2 precursor epitope 357Asp Gln Val Asp Val
Lys Asp Cys Ala Asn His Glu Ile Lys Lys 1 5 10 15
35820PRTArtificial SequenceDermatophagoides pteronyssinus Mite
group 2 allergen Der p 2 precursor epitope 358Asp Gln Val Asp Val
Lys Asp Cys Ala Asn His Glu Ile Lys Lys Val 1 5 10 15 Leu Val Pro
Gly 20 35915PRTArtificial SequenceDermatophagoides pteronyssinus
Mite group 2 allergen Der p 2 precursor epitope 359Cys His Gly Ser
Glu Pro Cys Ile Ile His Arg Gly Lys Pro Phe 1 5 10 15
36015PRTArtificial SequenceLepidoglyphus destructor Mite group 2
allergen
Lep d 2 precursor epitope 360Asp His Gly Val Met Ala Cys Gly Thr
Val His Gly Gln Val Glu 1 5 10 15 36115PRTArtificial
SequenceLepidoglyphus destructor Mite group 2 allergen Lep d 2
precursor epitope 361Gly Cys Lys Phe Ile Lys Cys Pro Val Lys Lys
Gly Glu Ala Leu 1 5 10 15 36215PRTArtificial SequenceLepidoglyphus
destructor Mite group 2 allergen Lep d 2 precursor epitope 362Gly
Glu Lys Met Thr Leu Glu Ala Lys Phe Ala Ala Asn Gln Asp 1 5 10 15
36315PRTArtificial SequenceLepidoglyphus destructor Mite group 2
allergen Lep d 2 precursor epitope 363Gly Glu Val Thr Glu Leu Asp
Ile Thr Gly Cys Ser Gly Asp Thr 1 5 10 15 36415PRTArtificial
SequenceLepidoglyphus destructor Mite group 2 allergen Lep d 2
precursor epitope 364Gly Lys Met Thr Phe Lys Asp Cys Gly His Gly
Glu Val Thr Glu 1 5 10 15 36516PRTArtificial SequenceHomo sapiens
Neurofilament heavy polypeptide (NF-H) (Neurofilament triplet H
protein) (200 kDa neurofilament protein) epitope 365Tyr Gln Glu Ala
Ile Gln Gln Leu Asp Ala Glu Leu Arg Asn Thr Lys 1 5 10 15
36610PRTArtificial SequencePrunus persica Non-specific
lipid-transfer protein 1 epitope 366Ala Ala Ala Leu Pro Gly Lys Cys
Gly Val 1 5 10 36710PRTArtificial SequencePrunus persica
Non-specific lipid-transfer protein 1 epitope 367Ala Cys Cys Asn
Gly Ile Arg Asn Val Asn 1 5 10 36810PRTArtificial SequencePrunus
persica Non-specific lipid-transfer protein 1 epitope 368Ala Pro
Cys Ile Pro Tyr Val Arg Gly Gly 1 5 10 36910PRTArtificial
SequencePrunus persica Non-specific lipid-transfer protein 1
epitope 369Ile Arg Asn Val Asn Asn Leu Ala Arg Thr 1 5 10
37011PRTArtificial SequencePrunus persica Non-specific
lipid-transfer protein 1 epitope 370Ile Ser Ala Ser Thr Asn Cys Ala
Thr Val Lys 1 5 10 37110PRTArtificial SequencePrunus persica
Non-specific lipid-transfer protein 1 epitope 371Asn Leu Ala Arg
Thr Thr Pro Asp Arg Gln 1 5 10 37210PRTArtificial SequenceGallus
gallus Ovalbumin epitope 372Cys Phe Asp Val Phe Lys Glu Leu Lys Val
1 5 10 37310PRTArtificial SequenceGallus gallus Ovalbumin epitope
373Gly Ser Ile Gly Ala Ala Ser Met Glu Phe 1 5 10
37418PRTArtificial SequenceGallus gallus Ovalbumin epitope 374Ile
Gly Leu Phe Arg Val Ala Ser Met Ala Ser Glu Lys Met Lys Ile 1 5 10
15 Leu Glu 37518PRTArtificial SequenceGallus gallus Ovalbumin
epitope 375Ile Lys His Ile Ala Thr Asn Ala Val Leu Phe Phe Gly Arg
Cys Val 1 5 10 15 Ser Pro 37613PRTArtificial SequenceGallus gallus
Ovalbumin epitope 376Ile Met Ser Ala Leu Ala Met Val Tyr Leu Gly
Ala Lys 1 5 10 37714PRTArtificial SequenceGallus gallus Ovomucoid
precursor epitope 377Ala Glu Val Asp Cys Ser Arg Phe Pro Asn Ala
Thr Asp Lys 1 5 10 37814PRTArtificial SequenceGallus gallus
Ovomucoid precursor epitope 378Ala Thr Asp Lys Glu Gly Lys Asp Val
Leu Val Cys Asn Lys 1 5 10 37917PRTArtificial SequenceGallus gallus
Ovomucoid precursor epitope 379Ala Val Val Glu Ser Asn Gly Thr Leu
Thr Leu Ser His Phe Gly Lys 1 5 10 15 Cys 38016PRTArtificial
SequenceGallus gallus Ovomucoid precursor epitope 380Cys Leu Leu
Cys Ala Tyr Ser Ile Glu Phe Gly Thr Asn Ile Ser Lys 1 5 10 15
38120PRTArtificial SequenceGallus gallus Ovomucoid precursor
epitope 381Asp Asn Glu Cys Leu Leu Cys Ala His Lys Val Glu Gln Gly
Ala Ser 1 5 10 15 Val Asp Lys Arg 20 38216PRTArtificial
SequenceMusa acuminata pectate lyase epitope 382Gly His Ser Asp Glu
Leu Thr Ser Asp Lys Ser Met Gln Val Thr Ile 1 5 10 15
38316PRTArtificial SequenceZinnia violacea Pectate lyase precursor
epitope 383Gly His Ser Asp Ser Tyr Thr Gln Asp Lys Asn Met Gln Val
Thr Ile 1 5 10 15 38421PRTArtificial SequenceDermatophagoides
farinae Peptidase 1 precursor (Major mite fecal allergen Der f 1)
(Allergen Der f I) epitope 384Asp Gly Arg Thr Ile Ile Gln His Asp
Asn Gly Tyr Gln Pro Asn Tyr 1 5 10 15 His Ala Val Asn Ile 20
38519PRTArtificial SequenceDermatophagoides farinae Peptidase 1
precursor (Major mite fecal allergen Der f 1) (Allergen Der f I)
epitope 385Asp Leu Arg Ser Leu Arg Thr Val Thr Pro Ile Arg Met Gln
Gly Gly 1 5 10 15 Cys Gly Ser 38619PRTArtificial
SequenceDermatophagoides farinae Peptidase 1 precursor (Major mite
fecal allergen Der f 1) (Allergen Der f I) epitope 386Gly Cys Gly
Ser Cys Trp Ala Phe Ser Gly Val Ala Ala Thr Glu Ser 1 5 10 15 Ala
Tyr Leu 38721PRTArtificial SequenceDermatophagoides farinae
Peptidase 1 precursor (Major mite fecal allergen Der f 1) (Allergen
Der f I) epitope 387Ile Arg Glu Ala Leu Thr Gln Thr His Thr Ala Ile
Ala Val Ile Ile 1 5 10 15 Gly Ile Lys Asp Leu 20 38819PRTArtificial
SequenceDermatophagoides farinae Peptidase 1 precursor (Major mite
fecal allergen Der f 1) (Allergen Der f I) epitope 388Ile Arg Met
Gln Gly Gly Cys Gly Ser Cys Trp Ala Phe Ser Gly Val 1 5 10 15 Ala
Ala Thr 38919PRTArtificial SequenceEuroglyphus maynei Peptidase 1
precursor (Mite group 1 allergen Eur m 1) (Allergen Eur m I)
epitope 389Phe Arg His Tyr Asp Gly Arg Thr Ile Met Gln His Asp Asn
Gly Tyr 1 5 10 15 Gln Pro Asn 39019PRTArtificial
SequenceEuroglyphus maynei Peptidase 1 precursor (Mite group 1
allergen Eur m 1) (Allergen Eur m I) epitope 390Gly Arg Thr Ile Met
Gln His Asp Asn Gly Tyr Gln Pro Asn Tyr His 1 5 10 15 Ala Val Asn
39119PRTArtificial SequenceEuroglyphus maynei Peptidase 1 precursor
(Mite group 1 allergen Eur m 1) (Allergen Eur m I) epitope 391His
Ala Val Asn Ile Val Gly Tyr Gly Asn Thr Gln Gly Val Asp Tyr 1 5 10
15 Trp Ile Val 39219PRTArtificial SequenceEuroglyphus maynei
Peptidase 1 precursor (Mite group 1 allergen Eur m 1) (Allergen Eur
m I) epitope 392Asn Lys Ile Arg Gln Ala Leu Thr Gln Thr His Thr Ala
Val Ala Val 1 5 10 15 Ile Ile Gly 39319PRTArtificial
SequenceEuroglyphus maynei Peptidase 1 precursor (Mite group 1
allergen Eur m 1) (Allergen Eur m I) epitope 393Pro Tyr Val Ala Arg
Glu Gln Ser Cys His Arg Pro Asn Ala Gln Arg 1 5 10 15 Tyr Gly Leu
39415PRTArtificial SequencePhleum pratense Phl p 3 allergen epitope
394Ala Val Gln Val Thr Phe Thr Val Gln Lys Gly Ser Asp Pro Lys 1 5
10 15 39515PRTArtificial SequencePhleum pratense Phl p 3 allergen
epitope 395Glu Glu Trp Glu Pro Leu Thr Lys Lys Gly Asn Val Trp Glu
Val 1 5 10 15 39615PRTArtificial SequencePhleum pratense Phl p 3
allergen epitope 396Phe Thr Val Gln Lys Gly Ser Asp Pro Lys Lys Leu
Val Leu Asp 1 5 10 15 39715PRTArtificial SequencePhleum pratense
Phl p 3 allergen epitope 397Phe Thr Val Gln Lys Gly Ser Asp Pro Lys
Lys Leu Val Leu Asn 1 5 10 15 39815PRTArtificial SequencePhleum
pratense Phl p 3 allergen epitope 398Gly Ser Asp Pro Lys Lys Leu
Val Leu Asp Ile Lys Tyr Thr Arg 1 5 10 15 39915PRTArtificial
SequenceApis mellifera Phospholipase A2 precursor epitope 399Cys
Asp Cys Asp Asp Lys Phe Tyr Asp Cys Leu Lys Asn Ser Ala 1 5 10 15
40012PRTArtificial SequenceApis mellifera Phospholipase A2
precursor epitope 400Cys Leu His Tyr Thr Val Asp Lys Ser Lys Pro
Lys 1 5 10 40115PRTArtificial SequenceApis mellifera Phospholipase
A2 precursor epitope 401Cys Arg Thr His Asp Met Cys Pro Asp Val Met
Ser Ala Gly Glu 1 5 10 15 40218PRTArtificial SequenceApis mellifera
Phospholipase A2 precursor epitope 402Asp Thr Ile Ser Ser Tyr Phe
Val Gly Lys Met Tyr Phe Asn Leu Ile 1 5 10 15 Asp Thr
40318PRTArtificial SequenceApis mellifera Phospholipase A2
precursor epitope 403Glu Arg Thr Glu Gly Arg Cys Leu His Tyr Thr
Val Asp Lys Ser Lys 1 5 10 15 Pro Lys 40416PRTArtificial
SequenceSpiroplasma citri plectrovirus spv1-r8a2b orf 14
transmembrane protein epitope 404His Val Ile Glu Val Gln Gln Ile
Asn Ser Glu Arg Ser Trp Phe Phe 1 5 10 15 40520PRTArtificial
SequenceLolium perenne pollen allergen epitope 405Cys Gly Tyr Lys
Asp Val Asp Lys Ala Pro Phe Asn Gly Met Thr Gly 1 5 10 15 Cys Gly
Asn Thr 20 40620PRTArtificial SequenceLolium perenne pollen
allergen epitope 406Gly Ala Gly Pro Lys Asp Asn Gly Gly Ala Cys Gly
Tyr Lys Asp Val 1 5 10 15 Asp Lys Ala Pro 20 40720PRTArtificial
SequenceLolium perenne pollen allergen epitope 407Ser Glu Val Glu
Asp Val Ile Pro Glu Gly Trp Lys Ala Asp Thr Ser 1 5 10 15 Tyr Ser
Ala Lys 20 40820PRTArtificial SequenceLolium perenne pollen
allergen epitope 408Val Glu Lys Gly Ser Asn Pro Asn Tyr Leu Ala Ile
Leu Val Lys Tyr 1 5 10 15 Val Asp Gly Asp 20 40920PRTArtificial
SequenceLolium perenne pollen allergen epitope 409Tyr Pro Asp Asp
Thr Lys Pro Thr Phe His Val Glu Lys Gly Ser Asn 1 5 10 15 Pro Asn
Tyr Leu 20 41016PRTArtificial SequenceAmbrosia artemisiifolia
Pollen allergen Amb a 1.1 precursor epitope 410Gly Ala Gly Asp Glu
Asn Ile Glu Asp Arg Gly Met Leu Ala Thr Val 1 5 10 15
41116PRTArtificial SequenceAmbrosia artemisiifolia Pollen allergen
Amb a 1.1 precursor epitope 411Gly Ala Gly Asp Glu Asn Ile Glu Asp
Arg Gly Met Leu Ala Thr Val 1 5 10 15 41216PRTArtificial
SequenceAmbrosia artemisiifolia Pollen allergen Amb a 2 precursor
epitope 412Gly Ala Ser Asp Thr His Phe Gln Asp Leu Lys Met His Val
Thr Leu 1 5 10 15 41316PRTArtificial SequenceAmbrosia
artemisiifolia Pollen allergen Amb a 2 precursor epitope 413Gly Ala
Ser Asp Thr His Phe Gln Asp Leu Lys Met His Val Thr Leu 1 5 10 15
41411PRTArtificial SequenceAmbrosia artemisiifolia var. elatior
Pollen allergen Amb a 3 epitope 414Glu Glu Ala Tyr His Ala Cys Asp
Ile Lys Asp 1 5 10 41515PRTArtificial SequenceAmbrosia
artemisiifolia var. elatior Pollen allergen Amb a 3 epitope 415Gly
Lys Val Tyr Leu Val Gly Gly Pro Glu Leu Gly Gly Trp Lys 1 5 10 15
41615PRTArtificial SequenceAmbrosia artemisiifolia var. elatior
Pollen allergen Amb a 3 epitope 416Leu Gly Gly Trp Lys Leu Gln Ser
Asp Pro Arg Ala Tyr Ala Leu 1 5 10 15 41715PRTArtificial
SequenceAmbrosia artemisiifolia var. elatior Pollen allergen Amb a
3 epitope 417Pro Gly Gly Pro Asp Arg Phe Thr Leu Leu Thr Pro Gly
Ser His 1 5 10 15 41815PRTArtificial SequenceAmbrosia
artemisiifolia var. elatior Pollen allergen Amb a 5 epitope 418Ala
Tyr Cys Cys Ser Asp Pro Gly Arg Tyr Cys Pro Trp Gln Val 1 5 10 15
41920PRTArtificial SequenceAmbrosia artemisiifolia var. elatior
Pollen allergen Amb a 5 epitope 419Cys Gly Glu Lys Arg Ala Tyr Cys
Cys Ser Asp Pro Gly Arg Tyr Cys 1 5 10 15 Pro Trp Gln Val 20
42017PRTArtificial SequenceAmbrosia artemisiifolia var. elatior
Pollen allergen Amb a 5 epitope 420Asp Pro Gly Arg Tyr Cys Pro Trp
Gln Val Val Cys Tyr Glu Ser Ser 1 5 10 15 Glu 42120PRTArtificial
SequenceAmbrosia artemisiifolia var. elatior Pollen allergen Amb a
5 epitope 421Asp Pro Gly Arg Tyr Cys Pro Trp Gln Val Val Cys Tyr
Glu Ser Ser 1 5 10 15 Glu Ile Cys Ser 20 42215PRTArtificial
SequenceAmbrosia artemisiifolia var. elatior Pollen allergen Amb a
5 epitope 422Gly Asn Val Cys Gly Glu Lys Arg Ala Tyr Cys Cys Ser
Asp Pro 1 5 10 15 42315PRTArtificial SequenceAmbrosia
artemisiifolia var. elatior Pollen allergen Amb a 5 epitope 423Leu
Val Pro Cys Ala Trp Ala Gly Asn Val Cys Gly Glu Lys Arg 1 5 10 15
42420PRTArtificial SequenceAmbrosia artemisiifolia var. elatior
Pollen allergen Amb a 5 epitope 424Leu Val Pro Cys Ala Trp Ala Gly
Asn Val Cys Gly Glu Lys Arg Ala 1 5 10 15 Tyr Cys Cys Ser 20
42515PRTArtificial SequenceAmbrosia artemisiifolia var. elatior
Pollen allergen Amb a 5 epitope 425Val Cys Tyr Glu Ser Ser Glu Ile
Cys Ser Lys Lys Cys Gly Lys 1 5 10 15 42620PRTArtificial
SequenceAmbrosia trifida Pollen allergen Amb t 5 precursor epitope
426Cys Gly Lys Val Gly Lys Tyr Cys Cys Ser Pro Ile Gly Lys Tyr Cys
1 5 10 15 Val Cys Tyr Asp 20 42720PRTArtificial SequenceAmbrosia
trifida Pollen allergen Amb t 5 precursor epitope 427Asp Asp Gly
Leu Cys Tyr Glu Gly Thr Asn Cys Gly Lys Val Gly Lys 1 5 10 15 Tyr
Cys Cys Ser 20 42812PRTArtificial SequenceAmbrosia trifida Pollen
allergen Amb t 5 precursor epitope 428Gly Lys Tyr Cys Val Cys Tyr
Asp Ser Lys Ala Ile 1 5 10 42914PRTArtificial SequenceAmbrosia
trifida Pollen allergen Amb t 5 precursor epitope 429Pro Ile Gly
Lys Tyr Cys Val Cys Tyr Asp Ser Lys Ala Ile 1 5 10
43020PRTArtificial SequenceAmbrosia trifida Pollen allergen Amb t 5
precursor epitope 430Pro Ile Gly Lys Tyr Cys Val Cys Tyr Asp Ser
Lys Ala Ile Cys Asn 1 5 10 15 Lys Asn Cys Thr 20 43114PRTArtificial
SequenceAmbrosia trifida Pollen allergen Amb t 5 precursor epitope
431Val Cys Tyr Asp Ser Lys Ala Ile Cys Asn Lys Asn Cys Thr 1 5 10
43215PRTArtificial SequenceBetula pendula pollen allergen Bet v 1
epitope 432His Glu Val Lys Ala Glu Gln Val Lys Ala Thr Lys Glu Met
Gly 1 5 10 15 43320PRTArtificial SequencePoa pratensis Pollen
allergen KBG 60 precursor epitope 433Ala Ala Asn Lys Tyr Lys Thr
Phe Val Ala Thr Phe Gly Ala Ala Ser 1 5 10 15 Asn Lys Ala Phe 20
43420PRTArtificial SequencePoa pratensis Pollen allergen KBG 60
precursor epitope 434Ala Ala Pro Ala Asn Asp Lys Phe Thr Val Phe
Glu Ala Ala Phe Asn 1 5 10 15 Asp Ala Ile Lys 20 43520PRTArtificial
SequencePoa pratensis Pollen allergen KBG 60 precursor epitope
435Ala Ala Val Asp Ser Ser Lys Ala Ala Leu Thr Ser Lys Leu Asp Ala
1 5 10 15 Ala Tyr Lys Leu 20 43620PRTArtificial SequencePoa
pratensis Pollen allergen KBG 60 precursor epitope 436Ala Glu Glu
Val Lys Ala Thr Pro Ala Gly Glu Leu Gln Val Ile Asp 1 5 10 15 Lys
Val Asp Ala 20 43720PRTArtificial SequencePoa pratensis Pollen
allergen KBG 60 precursor epitope 437Ala Phe Lys Val Ala Ala Thr
Ala Ala Asn Ala Ala Pro Ala Asn Asp 1 5 10 15 Lys Phe Thr Val 20
43820PRTArtificial SequenceLolium perenne Pollen allergen Lol p 1
precursor epitope 438Ala Phe Gly Ser Met Ala Lys Lys Gly Glu Glu
Gln Asn Val Arg Ser 1 5 10 15 Ala Gly Glu Leu 20 43920PRTArtificial
SequenceLolium perenne Pollen allergen Lol p 1
precursor epitope 439Ala Gly Glu Leu Glu Leu Gln Phe Arg Arg Val
Lys Cys Lys Tyr Pro 1 5 10 15 Asp Asp Thr Lys 20 44020PRTArtificial
SequenceLolium perenne Pollen allergen Lol p 1 precursor epitope
440Ala Lys Ser Thr Trp Tyr Gly Lys Pro Thr Gly Ala Gly Pro Lys Asp
1 5 10 15 Asn Gly Gly Ala 20 44120PRTArtificial SequenceLolium
perenne Pollen allergen Lol p 1 precursor epitope 441Ala Pro Tyr
His Phe Asp Leu Ser Gly His Ala Phe Gly Ser Met Ala 1 5 10 15 Lys
Lys Gly Glu 20 44212PRTArtificial SequenceLolium perenne Pollen
allergen Lol p 1 precursor epitope 442Ile Ala Pro Tyr His Phe Asp
Leu Ser Gly His Ala 1 5 10 44320PRTArtificial SequenceLolium
perenne Pollen allergen Lol p VA precursor epitope 443Ala Ala Leu
Thr Lys Ala Ile Thr Ala Met Thr Gln Ala Gln Lys Ala 1 5 10 15 Gly
Lys Pro Ala 20 44420PRTArtificial SequenceLolium perenne Pollen
allergen Lol p VA precursor epitope 444Ala Ala Asn Ala Ala Pro Thr
Asn Asp Lys Phe Thr Val Phe Glu Ser 1 5 10 15 Ala Phe Asn Lys 20
44520PRTArtificial SequenceLolium perenne Pollen allergen Lol p VA
precursor epitope 445Ala Asp Lys Phe Lys Ile Phe Glu Ala Ala Phe
Ser Glu Ser Ser Lys 1 5 10 15 Gly Leu Leu Ala 20 44620PRTArtificial
SequenceLolium perenne Pollen allergen Lol p VA precursor epitope
446Ala Phe Ser Glu Ser Ser Lys Gly Leu Leu Ala Thr Ser Ala Ala Lys
1 5 10 15 Ala Pro Gly Leu 20 44720PRTArtificial SequenceLolium
perenne Pollen allergen Lol p VA precursor epitope 447Ala Tyr Ala
Ala Thr Val Ala Ala Ala Pro Glu Val Lys Tyr Ala Val 1 5 10 15 Phe
Glu Ala Ala 20 44812PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 1 epitope 448Ala Cys Ser Gly Glu Pro Val Val Val His
Ile Thr 1 5 10 44912PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 1 epitope 449Ala Glu Asp Val Ile Pro Glu Gly Trp Lys
Ala Asp 1 5 10 45012PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 1 epitope 450Ala Gly Glu Leu Glu Leu Gln Phe Arg Arg
Val Lys 1 5 10 45112PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 1 epitope 451Asp Lys Trp Ile Glu Leu Lys Glu Ser Trp
Gly Ala 1 5 10 45212PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 1 epitope 452Asp Lys Trp Leu Asp Ala Lys Ser Thr Trp
Tyr Gly 1 5 10 45312PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 1 precursor epitope 453Phe Glu Ile Lys Cys Thr Lys
Pro Glu Ala Cys Ser 1 5 10 45412PRTArtificial SequencePhleum
pratense Pollen allergen Phl p 1 precursor epitope 454Tyr His Phe
Asp Leu Ser Gly His Ala Phe Gly Ala 1 5 10 45515PRTArtificial
SequencePhleum pratense Pollen allergen Phl p 1 precursor epitope
455Glu Leu Lys Glu Ser Trp Gly Ala Ile Trp Arg Ile Asp Thr Pro 1 5
10 15 45615PRTArtificial SequencePhleum pratense Pollen allergen
Phl p 1 precursor epitope 456Glu Pro Ile Ala Pro Tyr His Phe Asp
Leu Ser Gly His Ala Phe 1 5 10 15 45715PRTArtificial SequencePhleum
pratense Pollen allergen Phl p 1 precursor epitope 457Phe Glu Ile
Lys Cys Thr Lys Pro Glu Ala Cys Ser Gly Glu Pro 1 5 10 15
45815PRTArtificial SequencePhleum pratense Pollen allergen Phl p 1
precursor epitope 458Trp Gly Ala Ile Trp Arg Ile Asp Thr Pro Asp
Lys Leu Thr Gly 1 5 10 15 45915PRTArtificial SequencePhleum
pratense Pollen allergen Phl p 11 epitope 459Arg Tyr Ala Asn Pro
Ile Ala Phe Phe Arg Lys Glu Pro Leu Lys 1 5 10 15
46015PRTArtificial SequencePhleum pratense Pollen allergen Phl p 2
epitope 460Glu His Gly Ser Asp Glu Trp Val Ala Met Thr Lys Gly Glu
Gly 1 5 10 15 46115PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 2 epitope 461Glu Trp Val Ala Met Thr Lys Gly Glu Gly
Gly Val Trp Thr Phe 1 5 10 15 46215PRTArtificial SequencePhleum
pratense Pollen allergen Phl p 2 epitope 462Gly Val Trp Thr Phe Asp
Ser Glu Glu Pro Leu Gln Gly Pro Phe 1 5 10 15 46315PRTArtificial
SequencePhleum pratense Pollen allergen Phl p 2 epitope 463Lys Asn
Val Phe Asp Asp Val Val Pro Glu Lys Tyr Thr Ile Gly 1 5 10 15
46415PRTArtificial SequencePhleum pratense Pollen allergen Phl p 2
epitope 464Leu Gln Gly Pro Phe Asn Phe Arg Phe Leu Thr Glu Lys Gly
Met 1 5 10 15 46515PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 4 epitope 465Phe Lys Pro Phe Ala Glu Tyr Lys Ser Asp
Tyr Val Tyr Glu Pro 1 5 10 15 46615PRTArtificial SequencePhleum
pratense Pollen allergen Phl p 4 epitope 466Phe Pro Lys Glu Val Trp
Glu Gln Ile Phe Ser Thr Trp Leu Leu 1 5 10 15 46715PRTArtificial
SequencePhleum pratense Pollen allergen Phl p 4 epitope 467Phe Val
His Leu Gly His Arg Asp Asn Ile Glu Asp Asp Leu Leu 1 5 10 15
46815PRTArtificial SequencePhleum pratense Pollen allergen Phl p 4
epitope 468Gly Ile Val Val Ala Trp Lys Val Arg Leu Leu Pro Val Pro
Pro 1 5 10 15 46915PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 4 epitope 469Asn Arg Asn Asn Thr Phe Lys Pro Phe Ala
Glu Tyr Lys Ser Asp 1 5 10 15 47012PRTArtificial SequencePhleum
pratense Pollen allergen Phl p 5a epitope 470Glu Val Lys Tyr Thr
Val Phe Glu Thr Ala Leu Lys 1 5 10 47119PRTArtificial
SequencePhleum pratense Pollen allergen Phl p 5a epitope 471Asn Ala
Gly Phe Lys Ala Ala Leu Ala Gly Ala Gly Val Gln Pro Ala 1 5 10 15
Asp Lys Tyr 47227PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 5b precursor epitope 472Ala Ala Gly Lys Ala Thr Thr
Glu Glu Gln Lys Leu Ile Glu Asp Ile 1 5 10 15 Asn Val Gly Phe Lys
Ala Ala Val Ala Ala Ala 20 25 47333PRTArtificial SequencePhleum
pratense Pollen allergen Phl p 5b precursor epitope 473Ala Ala Gly
Lys Ala Thr Thr Glu Glu Gln Lys Leu Ile Glu Asp Ile 1 5 10 15 Asn
Val Gly Phe Lys Ala Ala Val Ala Ala Ala Ala Ser Val Pro Ala 20 25
30 Ala 47419PRTArtificial SequencePhleum pratense Pollen allergen
Phl p 5b precursor epitope 474Ala Ala Val Ala Ala Ala Ala Ser Val
Pro Ala Ala Asp Lys Phe Lys 1 5 10 15 Thr Phe Glu
47527PRTArtificial SequencePhleum pratense Pollen allergen Phl p 5b
precursor epitope 475Ala Lys Phe Asp Ser Phe Val Ala Ser Leu Thr
Glu Ala Leu Arg Val 1 5 10 15 Ile Ala Gly Ala Leu Glu Val His Ala
Val Lys 20 25 47619PRTArtificial SequencePhleum pratense Pollen
allergen Phl p 5b precursor epitope 476Ala Met Ser Glu Val Gln Lys
Val Ser Gln Pro Ala Thr Gly Ala Ala 1 5 10 15 Thr Val Ala
47720PRTArtificial SequenceChamaecyparis obtusa Polygalacturonase
epitope 477Ala Arg Trp Lys Asn Ser Lys Ile Trp Leu Gln Phe Ala Gln
Leu Thr 1 5 10 15 Asp Phe Asn Leu 20 47820PRTArtificial
SequenceChamaecyparis obtusa Polygalacturonase epitope 478Ala Val
Leu Leu Val Pro Ala Asn Lys Lys Phe Phe Val Asn Asn Leu 1 5 10 15
Val Phe Arg Gly 20 47920PRTArtificial SequenceChamaecyparis obtusa
Polygalacturonase epitope 479Asp Gly Thr Ile Val Ala Gln Pro Asp
Pro Ala Arg Trp Lys Asn Ser 1 5 10 15 Lys Ile Trp Leu 20
48020PRTArtificial SequenceChamaecyparis obtusa Polygalacturonase
epitope 480Phe Phe Val Asn Asn Leu Val Phe Arg Gly Pro Cys Gln Pro
His Leu 1 5 10 15 Ser Phe Lys Val 20 48120PRTArtificial
SequenceChamaecyparis obtusa Polygalacturonase epitope 481Phe Gly
Glu Cys Glu Gly Val Lys Ile Gln Gly Leu Lys Ile Lys Ala 1 5 10 15
Pro Arg Asp Ser 20 48215PRTArtificial SequenceCryptomeria japonica
Polygalacturonase precursor epitope 482Ala Ala Tyr Gln Asn Pro Ala
Ser Trp Lys Asn Asn Arg Ile Trp 1 5 10 15 48315PRTArtificial
SequenceCryptomeria japonica Polygalacturonase precursor epitope
483Ala Cys Lys Lys Pro Ser Ala Met Leu Leu Val Pro Gly Asn Lys 1 5
10 15 48415PRTArtificial SequenceCryptomeria japonica
Polygalacturonase precursor epitope 484Ala Ile Lys Phe Asp Phe Ser
Thr Gly Leu Ile Ile Gln Gly Leu 1 5 10 15 48515PRTArtificial
SequenceCryptomeria japonica Polygalacturonase precursor epitope
485Ala Ile Asn Ile Phe Asn Val Glu Lys Tyr Gly Ala Val Gly Asp 1 5
10 15 48615PRTArtificial SequenceCryptomeria japonica
Polygalacturonase precursor epitope 486Ala Asn Gly Tyr Phe Ser Gly
His Val Ile Pro Ala Cys Lys Asn 1 5 10 15 48716PRTArtificial
SequenceArabidopsis thaliana Probable pectate lyase 18 precursor
epitope 487Gly His Ser Asp Thr Tyr Ser Arg Asp Lys Asn Met Gln Val
Thr Ile 1 5 10 15 48815PRTArtificial SequencePhleum pratense
Profilin-2/4 epitope 488Leu Gly His Asp Gly Thr Val Trp Ala Gln Ser
Ala Asp Phe Pro 1 5 10 15 48920PRTArtificial SequenceHevea
brasiliensis Pro-hevein precursor epitope 489Asp Glu Tyr Cys Ser
Pro Asp His Asn Cys Gln Ser Asn Cys Lys Asp 1 5 10 15 Ser Gly Glu
Gly 20 49020PRTArtificial SequenceHevea brasiliensis Pro-hevein
precursor epitope 490Glu Gln Cys Gly Arg Gln Ala Gly Gly Lys Leu
Cys Pro Asn Asn Leu 1 5 10 15 Cys Cys Ser Gln 20 49143PRTArtificial
SequenceHevea brasiliensis Pro-hevein precursor epitope 491Glu Gln
Cys Gly Arg Gln Ala Gly Gly Lys Leu Cys Pro Asn Asn Leu 1 5 10 15
Cys Cys Ser Gln Trp Gly Trp Cys Gly Ser Thr Asp Glu Tyr Cys Ser 20
25 30 Pro Asp His Asn Cys Gln Ser Asn Cys Lys Asp 35 40
49220PRTArtificial SequenceHevea brasiliensis Pro-hevein precursor
epitope 492Lys Leu Cys Pro Asn Asn Leu Cys Cys Ser Gln Trp Gly Trp
Cys Gly 1 5 10 15 Ser Thr Asp Glu 20 49320PRTArtificial
SequenceHevea brasiliensis Pro-hevein precursor epitope 493Asn Gly
Gly Leu Asp Leu Asp Val Asn Val Phe Arg Gln Leu Asp Thr 1 5 10 15
Asp Gly Lys Gly 20 49410PRTArtificial SequencePrunus persica pru p
1 epitope 494Gly Lys Cys Gly Val Ser Ile Pro Tyr Lys 1 5 10
49510PRTArtificial SequencePrunus persica pru p 1 epitope 495Ile
Thr Cys Gly Gln Val Ser Ser Ser Leu 1 5 10 49610PRTArtificial
SequencePrunus persica pru p 1 epitope 496Ser Ile Pro Tyr Lys Ile
Ser Ala Ser Thr 1 5 10 49715PRTArtificial SequencePrunus persica
pru p 1 epitope 497Asp Arg Gln Ala Ala Cys Asn Cys Leu Lys Gln Leu
Ser Ala Ser 1 5 10 15 49815PRTArtificial SequencePrunus persica pru
p 1 epitope 498Val Asn Pro Asn Asn Ala Ala Ala Leu Pro Gly Lys Cys
Gly Val 1 5 10 15 49916PRTArtificial SequenceArabidopsis thaliana
Putative pectate lyase 17 precursor epitope 499Gly His Asn Asp Asn
Phe Val Lys Asp Val Lys Met Lys Val Thr Val 1 5 10 15
50016PRTArtificial SequenceHomo sapiens RAD51-like 1 isoform 1
epitope 500Thr Arg Leu Ile Leu Gln Tyr Leu Asp Ser Glu Arg Arg Gln
Ile Leu 1 5 10 15 50116PRTArtificial SequenceAspergillus fumigatus
Ribonuclease mitogillin precursor epitope 501Asp Pro Gly Pro Ala
Arg Val Ile Tyr Thr Tyr Pro Asn Lys Val Phe 1 5 10 15
50220PRTArtificial SequenceAspergillus fumigatus Ribonuclease
mitogillin precursor epitope 502Ala Thr Trp Thr Cys Ile Asn Gln Gln
Leu Asn Pro Lys Thr Asn Lys 1 5 10 15 Trp Glu Asp Lys 20
50320PRTArtificial SequenceAspergillus fumigatus Ribonuclease
mitogillin precursor epitope 503His Tyr Leu Leu Glu Phe Pro Thr Phe
Pro Asp Gly His Asp Tyr Lys 1 5 10 15 Phe Asp Ser Lys 20
50420PRTArtificial SequenceAspergillus fumigatus Ribonuclease
mitogillin precursor epitope 504Lys Phe Asp Ser Lys Lys Pro Lys Glu
Asp Pro Gly Pro Ala Arg Val 1 5 10 15 Ile Tyr Thr Tyr 20
50520PRTArtificial SequenceAspergillus fumigatus Ribonuclease
mitogillin precursor epitope 505Leu Ile Lys Gly Arg Thr Pro Ile Lys
Phe Gly Lys Ala Asp Cys Asp 1 5 10 15 Arg Pro Pro Lys 20
50620PRTArtificial SequenceAspergillus fumigatus Ribonuclease
mitogillin precursor epitope 506Ser Tyr Pro His Trp Phe Thr Asn Gly
Tyr Asp Gly Asn Gly Lys Leu 1 5 10 15 Ile Lys Gly Arg 20
50719PRTArtificial SequenceHevea brasiliensis Rubber elongation
factor protein epitope 507Ala Glu Asp Glu Asp Asn Gln Gln Gly Gln
Gly Glu Gly Leu Lys Tyr 1 5 10 15 Leu Gly Phe 50819PRTArtificial
SequenceHevea brasiliensis Rubber elongation factor protein epitope
508Phe Ser Asn Val Tyr Leu Phe Ala Lys Asp Lys Ser Gly Pro Leu Gln
1 5 10 15 Pro Gly Val 50919PRTArtificial SequenceHevea brasiliensis
Rubber elongation factor protein epitope 509Lys Phe Val Asp Ser Thr
Val Val Ala Ser Val Thr Ile Ile Asp Arg 1 5 10 15 Ser Leu Pro
51019PRTArtificial SequenceHevea brasiliensis Rubber elongation
factor protein epitope 510Gln Pro Gly Val Asp Ile Ile Glu Gly Pro
Val Lys Asn Val Ala Val 1 5 10 15 Pro Leu Tyr 51119PRTArtificial
SequenceHevea brasiliensis Rubber elongation factor protein epitope
511Arg Ser Leu Pro Pro Ile Val Lys Asp Ala Ser Ile Gln Val Val Ser
1 5 10 15 Ala Ile Arg 51217PRTArtificial SequenceBos taurus Serum
albumin precursor epitope 512Asp Asp Ser Pro Asp Leu Pro Lys Leu
Lys Pro Asp Pro Asn Thr Leu 1 5 10 15 Cys 51320PRTArtificial
SequenceBos taurus Serum albumin precursor epitope 513Glu Lys Asp
Ala Ile Pro Glu Asn Leu Pro Pro Leu Thr Ala Asp Phe 1 5 10 15 Ala
Glu Asp Lys 20 5149PRTArtificial SequenceBos taurus Serum albumin
precursor epitope 514Glu Ser His Ala Gly Cys Glu Lys Ser 1 5
51510PRTArtificial SequenceBos taurus Serum albumin precursor
epitope 515His Pro Glu Tyr Ala Val Ser Val Leu Leu 1 5 10
5169PRTArtificial SequenceBos taurus Serum albumin precursor
epitope 516Leu Ser Leu Ile Leu Asn Arg Leu Cys 1 5
51712PRTArtificial SequenceHevea brasiliensis Small rubber particle
protein epitope 517Asp Phe Val Arg Ala Ala Gly Val Tyr Ala Val Asp
1 5 10 51812PRTArtificial SequenceHevea brasiliensis Small rubber
particle protein epitope 518Lys Tyr Leu Asp Phe Val Arg Ala Ala Gly
Val Tyr 1 5 10 51912PRTArtificial SequenceHevea brasiliensis Small
rubber particle protein epitope 519Asn Val Val Lys Thr Val Val Thr
Pro Val Tyr Tyr 1 5 10 52012PRTArtificial SequenceHevea
brasiliensis Small rubber particle
protein epitope 520Pro Arg Ile Val Leu Asp Val Ala Ser Ser Val Phe
1 5 10 52112PRTArtificial SequenceHevea brasiliensis Small rubber
particle protein epitope 521Gln Gly Tyr Arg Val Ser Ser Tyr Leu Pro
Leu Leu 1 5 10 52215PRTArtificial SequenceGlycine max
Stress-induced protein SAM22 epitope 522Ala Leu Phe Lys Ala Ile Glu
Ala Tyr Leu Leu Ala His Pro Asp 1 5 10 15 52315PRTArtificial
SequenceCryptomeria japonica Sugi basic protein precursor epitope
523Ala Phe Asn Val Glu Asn Gly Asn Ala Thr Pro Gln Leu Thr Lys 1 5
10 15 52415PRTArtificial SequenceCryptomeria japonica Sugi basic
protein precursor epitope 524Ala Asn Asn Asn Tyr Asp Pro Trp Thr
Ile Tyr Ala Ile Gly Gly 1 5 10 15 52515PRTArtificial
SequenceCryptomeria japonica Sugi basic protein precursor epitope
525Ala Tyr Ser Asp Asp Lys Ser Met Lys Val Thr Val Ala Phe Asn 1 5
10 15 52615PRTArtificial SequenceCryptomeria japonica Sugi basic
protein precursor epitope 526Cys Gly Gln Arg Met Pro Arg Ala Arg
Tyr Gly Leu Val His Val 1 5 10 15 52715PRTArtificial
SequenceCryptomeria japonica Sugi basic protein precursor epitope
527Cys Ser Asn Trp Val Trp Gln Ser Thr Gln Asp Val Phe Tyr Asn 1 5
10 15 52820PRTArtificial SequenceTrichophyton rubrum Tri r 2
allergen epitope 528Ala Asp Phe Ser Asn Tyr Gly Ala Val Val Asp Val
Tyr Ala Pro Gly 1 5 10 15 Lys Asp Ile Thr 20 52920PRTArtificial
SequenceTrichophyton rubrum Tri r 2 allergen epitope 529Ala Lys Gly
Val Ser Leu Val Ala Val Lys Val Leu Asp Cys Asp Gly 1 5 10 15 Ser
Gly Ser Asn 20 53020PRTArtificial SequenceTrichophyton rubrum Tri r
2 allergen epitope 530Ala Ser Asn Gln Ala Ala Lys Ala Ile Ser Asp
Ala Gly Ile Phe Met 1 5 10 15 Ala Val Ala Ala 20 53120PRTArtificial
SequenceTrichophyton rubrum Tri r 2 allergen epitope 531Asp Cys Asn
Gly His Gly Thr His Val Ala Gly Thr Val Gly Gly Thr 1 5 10 15 Lys
Tyr Gly Leu 20 53220PRTArtificial SequenceTrichophyton rubrum Tri r
2 allergen epitope 532Asp Pro Ser Ala Gly Lys Gly Val Thr Ala Tyr
Ile Ile Asp Thr Gly 1 5 10 15 Ile Asp Ile Asp 20 53312PRTArtificial
SequenceVespula vulgaris Venom allergen 5 precursor epitope 533Ala
Cys Lys Tyr Gly Ser Leu Lys Pro Asn Cys Gly 1 5 10
53412PRTArtificial SequenceVespula vulgaris Venom allergen 5
precursor epitope 534Cys Asn Tyr Gly Pro Ser Gly Asn Phe Met Asn
Glu 1 5 10 53512PRTArtificial SequenceVespula vulgaris Venom
allergen 5 precursor epitope 535Asp Val Ala Lys Tyr Gln Val Gly Gln
Asn Val Ala 1 5 10 53612PRTArtificial SequenceVespula vulgaris
Venom allergen 5 precursor epitope 536Glu Lys Trp His Lys His Tyr
Leu Val Cys Asn Tyr 1 5 10 53712PRTArtificial SequenceVespula
vulgaris Venom allergen 5 precursor epitope 537Glu Leu Ala Tyr Val
Ala Gln Val Trp Ala Asn Gln 1 5 10 53815PRTArtificial
SequenceCorylus avellana 11S globulin-like protein epitope 538Ala
Phe Gln Ile Ser Arg Glu Glu Ala Arg Arg Leu Lys Tyr Asn 1 5 10 15
53912PRTArtificial SequenceCarya illinoinensis 11S legumin protein
epitope 539Glu Glu Ser Gln Arg Gln Ser Gln Gln Gly Gln Arg 1 5 10
54018PRTArtificial SequenceFagopyrum esculentum 13S globulin
epitope 540Asp Ala His Gln Pro Thr Arg Arg Val Arg Lys Gly Asp Val
Val Ala 1 5 10 15 Leu Pro 54112PRTArtificial SequenceFagopyrum
esculentum 13S globulin seed storage protein 1 precursor
(Legumin-like protein 1) epitope 541Phe Lys Gln Asn Val Asn Arg Pro
Ser Arg Ala Asp 1 5 10 54212PRTArtificial SequenceFagopyrum
esculentum 13S globulin seed storage protein 3 precursor
(Legumin-like protein 3) (Allergen Fag e 1) epitope 542Asp Ile Ser
Thr Lys Glu Ala Phe Arg Leu Lys Asn 1 5 10 54312PRTArtificial
SequenceAnacardium occidentale 2s albumin epitope 543Cys Gln Arg
Gln Phe Glu Glu Gln Gln Arg Phe Arg 1 5 10 54410PRTArtificial
SequenceSesamum indicum 2S seed storage protein 1 epitope 544His
Phe Arg Glu Cys Cys Asn Glu Ile Arg 1 5 10 54510PRTArtificial
SequenceSesamum indicum 2S seed storage protein 1 precursor epitope
545Cys Met Gln Trp Met Arg Ser Met Arg Gly 1 5 10
54614PRTArtificial SequenceBertholletia excelsa 2S sulfur-rich seed
storage protein precursor (Allergen Ber e 1) epitope 546Cys Arg Cys
Glu Gly Leu Arg Met Met Met Met Arg Met Gln 1 5 10
54740PRTArtificial SequenceHomo sapiens 52 kDa Ro protein epitope
547Leu Glu Lys Asp Glu Arg Glu Gln Leu Arg Ile Leu Gly Glu Lys Glu
1 5 10 15 Ala Lys Leu Ala Gln Gln Ser Gln Ala Leu Gln Glu Leu Ile
Ser Glu 20 25 30 Leu Asp Arg Arg Cys His Ser Ser 35 40
54810PRTArtificial SequenceHomo sapiens 52-kD SS-A/Ro autoantigen
epitope 548Gln Glu Lys Leu Gln Val Ala Leu Gly Glu 1 5 10
54921PRTArtificial SequenceHomo sapiens 5-hydroxytryptamine
(serotonin) receptor 4 epitope 549Gly Ile Ile Asp Leu Ile Glu Lys
Arg Lys Phe Asn Gln Asn Ser Asn 1 5 10 15 Ser Thr Tyr Cys Val 20
55010PRTArtificial SequenceHomo sapiens 60 kDa heat shock protein,
mitochondrial precursor epitope 550Asp Gly Val Ala Val Leu Lys Val
Gly Gly 1 5 10 55122PRTArtificial SequenceHomo sapiens 60 kDa
SS-A/Ro ribonucleoprotein epitope 551Glu Leu Tyr Lys Glu Lys Ala
Leu Ser Val Glu Thr Glu Lys Leu Leu 1 5 10 15 Lys Tyr Leu Glu Ala
Val 20 55222PRTArtificial SequenceHomo sapiens 60S acidic ribosomal
protein P0 epitope 552Ala Lys Val Glu Ala Lys Glu Glu Ser Glu Glu
Ser Asp Glu Asp Met 1 5 10 15 Gly Phe Gly Leu Phe Asp 20
55313PRTArtificial SequenceHomo sapiens 60S acidic ribosomal
protein P2 epitope 553Glu Glu Ser Asp Asp Asp Met Gly Phe Gly Leu
Phe Asp 1 5 10 55450PRTArtificial SequenceHomo sapiens 64 Kd
autoantigen epitope 554Ala Thr Lys Lys Glu Glu Glu Lys Lys Gly Gly
Asp Arg Asn Thr Gly 1 5 10 15 Leu Ser Arg Asp Lys Asp Lys Lys Arg
Glu Glu Met Lys Glu Val Ala 20 25 30 Lys Lys Glu Asp Asp Glu Lys
Val Lys Gly Glu Arg Arg Asn Thr Asp 35 40 45 Thr Arg 50
55519PRTArtificial SequenceHomo sapiens 65 kDa heat shock protein
epitope 555Ala Leu Leu Arg Cys Ile Pro Ala Leu Asp Ser Leu Thr Pro
Ala Asn 1 5 10 15 Glu Asp Cys 55614PRTArtificial SequenceHomo
sapiens Acetylcholine receptor subunit alpha precursor epitope
556Ala Ile Asn Pro Glu Ser Asp Gln Pro Asp Leu Ser Asn Phe 1 5 10
55721PRTArtificial SequenceCynodon dactylon acidic Cyn d 1
isoallergen isoform 1 precursor epitope 557Gln Asp Asp Val Ile Pro
Glu Asp Trp Lys Pro Asp Thr Val Tyr Lys 1 5 10 15 Ser Lys Ile Gln
Phe 20 55850PRTArtificial SequenceCynodon dactylon acidic Cyn d 1
isoallergen isoform 3 precursor epitope 558Glu Glu Asp Lys Leu Arg
Lys Ala Gly Glu Leu Met Leu Gln Phe Arg 1 5 10 15 Arg Val Lys Cys
Glu Tyr Pro Ser Asp Thr Lys Ile Thr Phe His Val 20 25 30 Glu Lys
Gly Ser Ser Pro Asn Tyr Leu Ala Leu Leu Val Lys Tyr Ala 35 40 45
Ala Gly 50 5598PRTArtificial SequenceHomo sapiens acidic ribosomal
phosphoprotein (P0) epitope 559Ala Ala Ala Ala Ala Pro Ala Lys 1 5
56015PRTArtificial SequenceHomo sapiens acidic ribosomal
phosphoprotein (P1) epitope 560Glu Ser Glu Glu Ser Asp Asp Asp Met
Gly Phe Gly Leu Phe Asp 1 5 10 15 56115PRTArtificial SequenceHomo
sapiens acidic ribosomal phosphoprotein (P2) epitope 561Ala Pro Ala
Ala Gly Ser Ala Pro Ala Ala Ala Glu Glu Lys Lys 1 5 10 15
56216PRTArtificial SequenceHomo sapiens Adrenergic, beta-2-,
receptor, surface epitope 562His Trp Tyr Arg Ala Thr His Gln Glu
Ala Ile Asn Cys Tyr Ala Asn 1 5 10 15 56310PRTArtificial
SequenceHomo sapiens Alanyl-tRNA synthetase, cytoplasmic epitope
563Phe Ile Asp Glu Pro Arg Arg Arg Pro Ile 1 5 10 5647PRTArtificial
SequenceBos taurus albumin epitope 564Pro Val Glu Ser Lys Val Thr 1
5 56513PRTArtificial SequenceJuglans regia Albumin seed storage
protein epitope 565Gly Leu Arg Gly Glu Glu Met Glu Glu Met Val Gln
Ser 1 5 10 56618PRTArtificial SequenceCochliobolus lunatus alcohol
dehydrogenase epitope 566Ala Val Asn Gly Asp Trp Pro Leu Pro Thr
Lys Leu Pro Leu Val Gly 1 5 10 15 Gly His 56737PRTArtificial
SequencePenicillium chrysogenum alkaline serine protease epitope
567Ala Asn Val Val Gln Arg Asn Ala Pro Ser Trp Gly Leu Ser Arg Ile
1 5 10 15 Ser Ser Lys Lys Ser Gly Ala Thr Asp Tyr Val Tyr Asp Ser
Thr Ala 20 25 30 Gly Glu Gly Ile Val 35 56810PRTArtificial
SequenceArachis hypogaea allergen epitope 568Asp Asp Gln Cys Gln
Arg Gln Leu Gln Arg 1 5 10 56915PRTArtificial SequenceAnacardium
occidentale allergen Ana o 2 epitope 569Glu Glu Ser Glu Asp Glu Lys
Arg Arg Trp Gly Gln Arg Asp Asn 1 5 10 15 57010PRTArtificial
SequenceArachis hypogaea Allergen Ara h 1, clone P41B precursor
epitope 570Ala Lys Ser Ser Pro Tyr Gln Lys Lys Thr 1 5 10
57115PRTArtificial SequenceArachis hypogaea allergen Arah3/Arah4
epitope 571Ala Gly Val Ala Leu Ser Arg Leu Val Leu Arg Arg Asn Ala
Leu 1 5 10 15 57210PRTArtificial SequenceArachis hypogaea allergen
Arah6 epitope 572Asp Arg Gln Met Val Gln His Phe Lys Arg 1 5 10
57311PRTArtificial SequencePeriplaneta americana Allergen Cr-PI
epitope 573Ile Pro Lys Gly Lys Lys Gly Gly Gln Ala Tyr 1 5 10
5748PRTArtificial SequenceAspergillus fumigatus allergen I/a; Asp f
I/a epitope 574Ile Asn Gln Gln Leu Asn Pro Lys 1 5
57510PRTArtificial SequenceArachis hypogaea Allergen II epitope
575Asp Arg Leu Gln Gly Arg Gln Gln Glu Gln 1 5 10
57615PRTArtificial SequenceLens culinaris allergen Len c 1.0101
epitope 576Ala Ile Asn Ala Ser Ser Asp Leu Asn Leu Ile Gly Phe Gly
Ile 1 5 10 15 57712PRTArtificial SequenceDermatophagoides farinae
Allergen Mag epitope 577Asp Val Glu Leu Ser Leu Arg Ser Ser Asp Ile
Ala 1 5 10 57831PRTArtificial SequencePenicillium chrysogenum
Allergen Pen n 18 epitope 578Ala His Ile Lys Lys Ser Lys Lys Gly
Asp Lys Lys Phe Lys Gly Ser 1 5 10 15 Val Ala Asn Met Ser Leu Gly
Gly Gly Ser Ser Arg Thr Leu Asp 20 25 30 57914PRTArtificial
SequenceSinapis alba Allergen Sin a 1 epitope 579Gln Gly Pro His
Val Ile Ser Arg Ile Tyr Gln Thr Ala Thr 1 5 10 58012PRTArtificial
SequenceZiziphus mauritiana allergen Ziz m 1 epitope 580Lys Thr Asn
Tyr Ser Ser Ser Ile Ile Leu Glu Tyr 1 5 10 58137PRTArtificial
SequenceFagopyrum tataricum allergenic protein epitope 581Asp Ile
Ser Thr Glu Glu Ala Tyr Lys Leu Lys Asn Gly Arg Gln Glu 1 5 10 15
Val Glu Val Phe Arg Pro Phe Gln Ser Arg Tyr Glu Lys Glu Glu Glu 20
25 30 Lys Glu Arg Glu Arg 35 58214PRTArtificial SequenceHomo
sapiens alpha 2 interferon epitope 582Glu Val Val Arg Ala Glu Ile
Met Arg Ser Phe Ser Leu Ser 1 5 10 58330PRTArtificial SequenceBos
taurus alpha S1 casein epitope 583Glu Asp Gln Ala Met Glu Asp Ile
Lys Gln Met Glu Ala Glu Ser Ile 1 5 10 15 Ser Ser Ser Glu Glu Ile
Val Pro Asn Ser Val Glu Gln Lys 20 25 30 58410PRTArtificial
SequenceTriticum aestivum Alpha/beta-gliadin A-II precursor epitope
584Gln Val Ser Phe Gln Gln Pro Gln Gln Gln 1 5 10
58510PRTArtificial SequenceTriticum aestivum Alpha/beta-gliadin A-V
epitope 585Leu Ala Leu Gln Thr Leu Pro Ala Met Cys 1 5 10
58610PRTArtificial SequenceHomo sapiens alpha-1 type IV collagen
epitope 586Ser Arg Cys Gln Val Cys Met Arg Arg Thr 1 5 10
58712PRTArtificial SequenceHomo sapiens alpha1A-voltage-dependent
calcium channel epitope 587Glu Asp Ser Asp Glu Asp Glu Phe Gln Ile
Thr Glu 1 5 10 58815PRTArtificial SequenceHomo sapiens alpha-2 type
XI collagen epitope 588Gly Ser Leu Asp Ser Leu Arg Arg Glu Ile Glu
Gln Met Arg Arg 1 5 10 15 58920PRTArtificial SequenceBos taurus
alpha2(I) collagen epitope 589Leu Pro Gly Leu Lys Gly His Asn Gly
Leu Gln Gly Leu Pro Gly Leu 1 5 10 15 Ala Gly His His 20
5905PRTArtificial SequenceTriticum aestivum Alpha-amylase inhibitor
0.28 precursor (CIII) (WMAI-1) epitope 590Ala Tyr Pro Asp Val 1 5
59117PRTArtificial SequenceHomo sapiens Alpha-enolase epitope
591Lys Ile His Ala Arg Glu Ile Phe Asp Ser Arg Gly Asn Pro Thr Val
1 5 10 15 Glu 59215PRTArtificial SequenceHomo sapiens
alpha-fibrinogen precursor epitope 592Gly Pro Arg Val Val Glu Arg
His Gln Ser Ala Cys Lys Asp Ser 1 5 10 15 59312PRTArtificial
SequenceTriticum aestivum Alpha-gliadin epitope 593Leu Gly Gln Gly
Ser Phe Arg Pro Ser Gln Gln Asn 1 5 10 59410PRTArtificial
SequenceBos taurus Alpha-lactalbumin epitope 594Lys Asp Leu Lys Gly
Tyr Gly Gly Val Ser 1 5 10 59514PRTArtificial SequenceBos taurus
Alpha-lactalbumin precursor epitope 595Lys Cys Glu Val Phe Arg Glu
Leu Lys Asp Leu Lys Gly Tyr 1 5 10 59620PRTArtificial SequenceBos
taurus alpha-S1-casein epitope 596Leu Asn Glu Asn Leu Leu Arg Phe
Phe Val Ala Pro Phe Pro Gln Val 1 5 10 15 Phe Gly Lys Glu 20
59710PRTArtificial SequenceBos taurus Alpha-S1-casein precursor
epitope 597Ala Met Glu Asp Ile Lys Gln Met Glu Ala 1 5 10
59810PRTArtificial SequenceBos taurus Alpha-S2-casein precursor
epitope 598Glu Asn Leu Cys Ser Thr Phe Cys Lys Glu 1 5 10
5997PRTArtificial SequenceHomo sapiens anti-beta-amyloid peptide
immunoglobulin heavy chain variable region epitope 599Ala His Ile
Trp Trp Asn Asp 1 5 60024PRTArtificial SequenceHomo sapiens
Aquaporin-4 epitope 600Phe Cys Pro Asp Val Glu Phe Lys Arg Arg Phe
Lys Glu Ala Phe Ser 1 5 10 15 Lys Ala Ala Gln Gln Thr Lys Gly 20
60115PRTArtificial SequenceArachis hypogaea Ara h 2.01 allergen
epitope 601Cys Cys Asn Glu Leu Asn Glu Phe Glu Asn Asn Gln Arg Cys
Met 1 5 10 15 60220PRTArtificial SequenceHomo sapiens ATP-dependent
DNA helicase 2 subunit 2 epitope 602Glu Glu Ala Ser Gly Ser Ser Val
Thr Ala Glu Glu Ala Lys Lys Phe 1 5 10 15 Leu Ala Pro Lys 20
60328PRTArtificial SequenceHomo sapiens autoantigen epitope 603Glu
Ile Arg Val Arg Leu Gln Ser Ala Ser Pro Ser Thr Arg Trp Thr 1 5 10
15 Glu Leu Asp Asp Val Lys Arg Leu Leu Lys Gly Ser 20 25
60416PRTArtificial SequenceHomo sapiens Band 3 anion transport
protein epitope 604Leu
Phe Lys Pro Pro Lys Tyr His Pro Asp Val Pro Tyr Val Lys Arg 1 5 10
15 60515PRTArtificial SequenceGlycine max Bd 30K (34 kDa maturing
seed protein) epitope 605Glu Asp Trp Gly Glu Asp Gly Tyr Ile Trp
Ile Gln Arg Asn Thr 1 5 10 15 6067PRTArtificial SequenceHomo
sapiens Bence Jones protein HAG epitope 606Ala Trp His Gln Gln Gln
Pro 1 5 6078PRTArtificial SequenceBetula pendula Bet v 4 epitope
607Phe Ala Arg Ala Asn Arg Gly Leu 1 5 6089PRTArtificial
SequenceMusa acuminata beta-1, 3-glucananse epitope 608Gly Leu Phe
Tyr Pro Asn Lys Gln Pro 1 5 60915PRTArtificial SequenceHevea
brasiliensis beta-1,3-glucanase epitope 609Gly Leu Phe Phe Pro Asp
Lys Arg Pro Lys Tyr Asn Leu Asn Phe 1 5 10 15 61012PRTArtificial
SequenceOlea europaea beta-1,3-glucanase-like protein epitope
610Ala Gly Arg Asn Ser Trp Asn Cys Asp Phe Ser Gln 1 5 10
6116PRTArtificial SequenceHomo sapiens beta-2-glycoprotein 1
precursor epitope 611Leu Lys Thr Pro Arg Val 1 5 6126PRTArtificial
SequenceHomo sapiens beta-2-glycoprotein I epitope 612Thr Leu Arg
Val Tyr Lys 1 5 61313PRTArtificial SequenceBos taurus beta-casein
epitope 613Gln Ser Lys Val Leu Pro Val Pro Gln Lys Ala Val Pro 1 5
10 61412PRTArtificial SequenceBos taurus Beta-casein precursor
epitope 614Asp Glu Leu Gln Asp Lys Ile His Pro Phe Ala Gln 1 5 10
61510PRTArtificial SequenceBos taurus Beta-lactoglobulin epitope
615Ala Gln Lys Lys Ile Ile Ala Glu Lys Thr 1 5 10
61616PRTArtificial SequenceBos taurus Beta-lactoglobulin precursor
epitope 616Ala Ala Ser Asp Ile Ser Leu Leu Asp Ala Gln Ser Ala Pro
Leu Arg 1 5 10 15 6177PRTArtificial SequenceHomo sapiens Botulinum
neurotoxin type E epitope 617Trp Lys Ala Pro Ser Ser Pro 1 5
61819PRTArtificial SequenceHomo sapiens bullous pemphigoid antigen
epitope 618Lys Ser Thr Ala Lys Asp Cys Thr Phe Lys Pro Asp Phe Glu
Met Thr 1 5 10 15 Val Lys Glu 61920PRTArtificial SequenceHomo
sapiens Bullous pemphigoid antigen 1, isoforms 1/2/3/4/5/8 epitope
619Leu Thr Asp Thr Lys Thr Gly Leu His Phe Asn Ile Asn Glu Ala Ile
1 5 10 15 Glu Gln Gly Thr 20 6208PRTArtificial SequenceFagopyrum
esculentum BW 16kDa allergen epitope 620Glu Gly Val Arg Asp Leu Lys
Glu 1 5 62117PRTArtificial SequenceHomo sapiens calcium channel,
alpha 1A subunit isoform 3 epitope 621Gly Asn Ile Gly Ile Asp Val
Glu Asp Glu Asp Ser Asp Glu Asp Glu 1 5 10 15 Phe
62215PRTArtificial SequenceHomo sapiens Calpastatin epitope 622Ala
Val Cys Arg Thr Ser Met Cys Ser Ile Gln Ser Ala Pro Pro 1 5 10 15
62318PRTArtificial SequenceHomo sapiens Calreticulin precursor
epitope 623Lys Glu Gln Phe Leu Asp Gly Asp Gly Trp Thr Ser Arg Trp
Ile Glu 1 5 10 15 Ser Lys 62413PRTArtificial SequenceHomo sapiens
Ca-sensing receptor epitope 624Phe Val Ala Gln Asn Lys Ile Asp Ser
Leu Asn Leu Asp 1 5 10 6259PRTArtificial SequenceHomo sapiens
Caspase-8 precursor epitope 625Asp Arg Asn Gly Thr His Leu Asp Ala
1 5 62614PRTArtificial SequenceHomo sapiens centromere protein A
isoform a epitope 626Gly Pro Ser Arg Arg Gly Pro Ser Leu Gly Ala
Ser Ser His 1 5 10 62710PRTArtificial SequenceHomo sapiens
centromere protein B, 80kDa epitope 627Met Gly Pro Lys Arg Arg Gln
Leu Thr Phe 1 5 10 62810PRTArtificial SequenceHomo sapiens
centromere protein-A epitope 628Glu Ala Pro Arg Arg Arg Ser Pro Ser
Pro 1 5 10 62917PRTArtificial SequenceBetula pendula Chain A, Birch
Pollen Profilin epitope 629Ala Gln Ser Ser Ser Phe Pro Gln Phe Lys
Pro Gln Glu Ile Thr Gly 1 5 10 15 Ile 6305PRTArtificial
SequenceHomo sapiens Chain A, Crystal Structure Of The Glycosylated
Five-Domain Human Beta2-Glycoprotein I Purified From Blood Plasma
epitope 630Arg Gly Gly Met Arg 1 5 6317PRTArtificial SequenceHomo
sapiens Chain H, Three-Dimensional Structure Of A Human
Immunoglobulin With A Hinge Deletion epitope 631Ala Leu Pro Ala Pro
Ile Glu 1 5 63227PRTArtificial SequenceHomo sapiens cholesterol
side-chain cleavage enzyme P450scc (EC 1.14.15.67) epitope 632Phe
Asp Pro Glu Asn Phe Asp Pro Thr Arg Trp Leu Ser Lys Asp Lys 1 5 10
15 Asn Ile Thr Tyr Phe Arg Asn Leu Gly Phe Gly 20 25
63310PRTArtificial SequenceHomo sapiens citrate synthase epitope
633Ala Leu Lys His Leu Pro Asn Asp Pro Met 1 5 10 6345PRTArtificial
SequenceHomo sapiens claudin 11 epitope 634Ala His Arg Glu Thr 1 5
63511PRTArtificial SequenceHomo sapiens Coagulation factor VIII
precursor epitope 635Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr 1
5 10 63620PRTArtificial SequenceOncorhynchus mykiss collagen a2(I)
epitope 636Met Lys Gly Leu Arg Gly His Gly Gly Leu Gln Gly Met Pro
Gly Pro 1 5 10 15 Asn Gly Pro Ser 20 63714PRTArtificial
SequenceHomo sapiens Collagen alpha-1(II) chain epitope 637Ala Arg
Gly Ala Gln Gly Pro Pro Gly Ala Thr Gly Phe Pro 1 5 10
6388PRTArtificial SequenceHomo sapiens collagen alpha-1(VII) chain
precursor epitope 638Gly Thr Leu His Val Val Gln Arg 1 5
63923PRTArtificial SequenceHomo sapiens Collagen alpha-1(XVII)
chain epitope 639Arg Ser Ile Leu Pro Tyr Gly Asp Ser Met Asp Arg
Ile Glu Lys Asp 1 5 10 15 Arg Leu Gln Gly Met Ala Pro 20
64015PRTArtificial SequenceHomo sapiens Collagen alpha-3(IV) chain
epitope 640Thr Ala Ile Pro Ser Cys Pro Glu Gly Thr Val Pro Leu Tyr
Ser 1 5 10 15 6418PRTArtificial SequenceHomo sapiens collagen VII
epitope 641Ile Ile Trp Arg Ser Thr Gln Gly 1 5 64246PRTArtificial
SequenceBos taurus collagen, type I, alpha 2 epitope 642Ala Pro Gly
Pro Asp Gly Asn Asn Gly Ala Gln Gly Pro Pro Gly Leu 1 5 10 15 Gln
Gly Val Gln Gly Gly Lys Gly Glu Gln Gly Pro Ala Gly Pro Pro 20 25
30 Gly Phe Gln Gly Leu Pro Gly Pro Ala Gly Thr Ala Gly Glu 35 40 45
6439PRTArtificial SequenceHomo sapiens collagen, type II, alpha 1
epitope 643Pro Pro Gly Pro Thr Gly Ala Ser Gly 1 5
64411PRTArtificial SequenceHomo sapiens collagen, type II, alpha 1
isoform 1 precursor epitope 644Ala Arg Gly Leu Thr Gly Arg Pro Gly
Asp Ala 1 5 10 64511PRTArtificial SequenceHomo sapiens collagen,
type II, alpha 1 isoform 2 precursor epitope 645Leu Val Gly Pro Arg
Gly Glu Arg Gly Phe Pro 1 5 10 6469PRTArtificial SequenceHomo
sapiens Complement C1q subcomponent subunit A epitope 646Lys Gly
Glu Gln Gly Glu Pro Gly Ala 1 5 64710PRTArtificial SequenceHomo
sapiens Condensin-2 complex subunit D3 epitope 647Pro Thr Pro Glu
Thr Gly Pro Leu Gln Arg 1 5 10 64815PRTArtificial SequenceArachis
hypogaea Conglutin-7 precursor epitope 648Ala Ala His Ala Ser Ala
Arg Gln Gln Trp Glu Leu Gln Gly Asp 1 5 10 15 6498PRTArtificial
SequencePeriplaneta americana Cr-PII allergen epitope 649Ile Arg
Ser Trp Phe Gly Leu Pro 1 5 65011PRTArtificial SequenceCochliobolus
lunatus Cytochrome c epitope 650Glu Asn Pro Lys Lys Tyr Ile Pro Gly
Thr Lys 1 5 10 65110PRTArtificial SequenceRattus norvegicus
Cytochrome P450 3A1 epitope 651Asp Met Val Leu Asn Glu Thr Leu Arg
Leu 1 5 10 65215PRTArtificial SequenceHomo sapiens
cytoskeleton-associated protein 5 isoform b epitope 652Cys Gln Ala
Leu Val Arg Met Leu Ala Lys Lys Pro Gly Trp Lys 1 5 10 15
6539PRTArtificial SequenceDermatophagoides farinae Der f 2 epitope
653Ile Ala Thr His Ala Lys Ile Arg Asp 1 5 65415PRTArtificial
SequenceDermatophagoides farinae Der f 7 allergen epitope 654His
Ile Gly Gly Leu Ser Ile Leu Asp Pro Ile Phe Gly Val Leu 1 5 10 15
65543PRTArtificial SequenceDermatophagoides pteronyssinus Der p 1
allergen epitope 655Ala Arg Glu Gln Ser Cys Arg Arg Pro Asn Ala Gln
Arg Phe Gly Ile 1 5 10 15 Ser Asn Tyr Cys Gln Ile Tyr Pro Pro Asn
Ala Asn Lys Ile Arg Glu 20 25 30 Ala Leu Ala Gln Thr His Ser Ala
Ile Ala Val 35 40 65615PRTArtificial SequenceDermatophagoides
pteronyssinus Der p 7 allergen polypeptide epitope 656His Ile Gly
Gly Leu Ser Ile Leu Asp Pro Ile Phe Ala Val Leu 1 5 10 15
65712PRTArtificial SequenceHomo sapiens Desmoglein-1 epitope 657Arg
Glu Trp Ile Lys Phe Ala Ala Ala Cys Arg Glu 1 5 10
65812PRTArtificial SequenceHomo sapiens Desmoglein-3 precursor
epitope 658Arg Glu Trp Val Lys Phe Ala Lys Pro Cys Arg Glu 1 5 10
65930PRTArtificial SequenceHomo sapiens desmoglein-3 preproprotein
epitope 659Ser Gln Glu Pro Ala Gly Thr Pro Met Phe Leu Leu Ser Arg
Asn Thr 1 5 10 15 Gly Glu Val Arg Thr Leu Thr Asn Ser Leu Asp Arg
Glu Gln 20 25 30 66012PRTArtificial SequenceHomo sapiens
desmoplakin epitope 660Gly Asn Ser Ser Tyr Ser Tyr Ser Tyr Ser Phe
Ser 1 5 10 66120PRTArtificial SequenceHomo sapiens desmoplakin
isoform II epitope 661Leu Val Asp Arg Lys Thr Gly Ser Gln Tyr Asp
Ile Gln Asp Ala Ile 1 5 10 15 Asp Lys Gly Leu 20 66218PRTArtificial
SequenceHomo sapiens dihydrolipoamide S-acetyltransferase (E2
component of pyruvate dehydrogenase complex), isoform CRA_a epitope
662Ala Glu Ile Glu Thr Asp Lys Ala Thr Ile Gly Phe Glu Val Gln Glu
1 5 10 15 Glu Gly 66317PRTArtificial SequenceHomo sapiens DNA
topoisomerase 1 epitope 663Gly Val Pro Ile Glu Lys Ile Tyr Asn Lys
Thr Gln Arg Glu Lys Phe 1 5 10 15 Ala 66420PRTArtificial
SequenceHomo sapiens DNA topoisomerase I epitope 664Glu Leu Asp Gly
Gln Glu Tyr Val Val Glu Phe Asp Phe Leu Gly Lys 1 5 10 15 Asp Ser
Ile Arg 20 66520PRTArtificial SequenceHomo sapiens DNA
topoisomerase II beta epitope 665His Pro Met Leu Pro Asn Tyr Lys
Asn Phe Lys Gly Thr Ile Gln Glu 1 5 10 15 Leu Gly Gln Asn 20
6667PRTArtificial SequenceHomo sapiens DNA-directed RNA polymerase
II subunit RPB1 epitope 666Tyr Ser Pro Thr Ser Pro Ser 1 5
66736PRTArtificial SequenceHomo sapiens E3 ubiquitin-protein ligase
TRIM9 isoform 2 epitope 667Ala Phe Asn Lys Thr Gly Val Ser Pro Tyr
Ser Lys Thr Leu Val Leu 1 5 10 15 Gln Thr Ser Glu Gly Lys Ala Leu
Gln Gln Tyr Pro Ser Glu Arg Glu 20 25 30 Leu Arg Gly Ile 35
66839PRTArtificial SequenceCandida albicans Enolase 1
(2-phosphoglycerate dehydratase) (2-phospho-D-glycerate
hydro-lyase) epitope 668Gln Ala Ala Asn Asp Ser Tyr Ala Ala Gly Trp
Gly Val Met Val Ser 1 5 10 15 His Arg Ser Gly Glu Thr Glu Asp Thr
Phe Ile Ala Asp Leu Ser Val 20 25 30 Gly Leu Arg Ser Gly Gln Ile 35
66916PRTArtificial SequenceHomo sapiens enolase 1 variant epitope
669Lys Ile His Ala Arg Glu Ile Phe Asp Ser Arg Gly Asn Pro Thr Val
1 5 10 15 67021PRTArtificial SequenceHevea brasiliensis ENSP-like
protein epitope 670Phe Pro Leu Ile Thr Cys Cys Gly Tyr Gly Gly Lys
Tyr Asn Phe Ser 1 5 10 15 Val Thr Ala Pro Cys 20 67120PRTArtificial
SequenceHomo sapiens envoplakin epitope 671Ala Gly Glu Thr Lys Pro
Ser Ser Ser Leu Ser Ile Gly Ser Ile Ile 1 5 10 15 Ser Lys Ser Pro
20 67212PRTArtificial SequenceFagopyrum esculentum Fag e 1 epitope
672Ala Val Val Leu Lys Ala Gly Asn Glu Gly Leu Glu 1 5 10
6733PRTArtificial SequenceHomo sapiens Fas AMA epitope 673Cys Val
Pro 1 67418PRTArtificial SequenceHomo sapiens FGA protein epitope
674Ser Arg Ala Leu Ala Arg Glu Val Asp Leu Lys Asp Tyr Glu Asp Gln
1 5 10 15 Gln Lys 67520PRTArtificial SequenceHomo sapiens FGB
protein epitope 675Ala Arg Gly His Arg Pro Leu Asp Lys Lys Arg Glu
Glu Ala Pro Ser 1 5 10 15 Leu Arg Pro Ala 20 67619PRTArtificial
SequenceHomo sapiens fibrin beta epitope 676Ala Asn Lys Tyr Gln Ile
Ser Val Asn Lys Tyr Arg Gly Thr Ala Gly 1 5 10 15 Asn Ala Leu
67715PRTArtificial SequenceHomo sapiens fibrinogen alpha chain
isoform alpha preproprotein epitope 677Asp Ser Pro Gly Ser Gly Asn
Ala Arg Pro Asn Asn Pro Asp Trp 1 5 10 15 67817PRTArtificial
SequenceHomo sapiens Fibrinogen alpha chain precursor epitope
678Phe Leu Ala Glu Gly Gly Gly Val Arg Gly Pro Arg Val Val Glu Arg
1 5 10 15 His 67920PRTArtificial SequenceHomo sapiens fibrinogen
alpha chain preproprotein, isoform alpha epitope 679Asp His Glu Gly
Thr His Ser Thr Lys Arg Gly His Ala Lys Ser Arg 1 5 10 15 Pro Val
Arg Gly 20 68015PRTArtificial SequenceHomo sapiens fibrinogen beta
chain epitope 680Pro Arg Lys Gln Cys Ser Lys Glu Asp Gly Gly Gly
Trp Trp Tyr 1 5 10 15 68117PRTArtificial SequenceHomo sapiens
fibrinogen beta chain, isoform CRA_d epitope 681Asn Glu Glu Gly Phe
Phe Ser Ala Arg Gly His Arg Pro Leu Asp Lys 1 5 10 15 Lys
68224PRTArtificial SequenceHomo sapiens fibrinogen beta chain,
isoform CRA_i epitope 682Glu Glu Ala Pro Ser Leu Arg Pro Ala Pro
Pro Pro Ile Ser Gly Gly 1 5 10 15 Gly Tyr Arg Ala Arg Pro Ala Lys
20 6836PRTArtificial SequenceHomo sapiens Fibronectin precursor
epitope 683Leu Thr Ser Arg Pro Ala 1 5 68418PRTArtificial
SequenceHomo sapiens filaggrin epitope 684Asp Ser Gly His Arg Gly
Tyr Ser Gly Ser Gln Ala Ser Asp Asn Glu 1 5 10 15 Gly His
6858PRTArtificial SequenceHomo sapiens Follistatin-related protein
1 epitope 685Leu Lys Phe Val Glu Gln Asn Glu 1 5 68610PRTArtificial
SequenceHomo sapiens Forkhead box protein E3 epitope 686Pro Thr Pro
Ala Pro Gly Pro Gly Arg Arg 1 5 10 6877PRTArtificial SequenceHomo
sapiens GAD65 autoantigen glutamic acid decarboxylase epitope
687Ala Pro Ala Met Ile Pro Pro 1 5 68810PRTArtificial
SequenceTriticum aestivum Gamma-gliadin precursor epitope 688Leu
Gln Pro Gln Gln Pro Phe Pro Gln Gln 1 5 10 68921PRTArtificial
SequenceChironomus thummi thummi Globin CTT-III epitope 689Ala His
Thr Asp Phe Ala Gly Ala Glu Ala Ala Trp Gly Ala Thr Leu 1 5 10 15
Asp Thr Phe Phe Gly 20 69011PRTArtificial SequenceChironomus thummi
thummi Globin CTT-III precursor epitope 690Gly Val Thr His Asp Gln
Leu Asn Asn Phe Arg 1 5 10 69123PRTArtificial SequenceChironomus
thummi thummi Globin CTT-IV precursor epitope 691Lys Ala His Thr
Asp Phe Ala Gly Ala Glu Ala Ala Trp Gly Ala Thr 1 5 10 15 Leu Asp
Ala Phe Phe Gly Met 20 69235PRTArtificial SequenceChironomus thummi
thummi Globin CTT-VI precursor epitope 692Ile Val Ser Phe Leu Ser
Glu Val Ile Ser Leu Ala Gly Ser Asp Ala 1 5 10 15 Asn Ile Pro Ala
Ile Gln Asn Leu Ala Lys Glu Leu Ala Thr Ser His
20 25 30 Lys Pro Arg 35 69335PRTArtificial SequenceChironomus
thummi thummi Globin CTT-VIII epitope 693Ile Val Gly Phe Phe Ser
Glu Val Ile Gly Leu Ile Gly Asn Pro Glu 1 5 10 15 Asn Arg Pro Ala
Leu Lys Thr Leu Ile Asp Gly Leu Ala Ser Ser His 20 25 30 Lys Ala
Arg 35 6949PRTArtificial SequenceHevea brasiliensis Glucan
endo-1,3-beta-glucosidase, basic vacuolar isoform epitope 694Ala
Trp Leu Ala Gln Phe Val Leu Pro 1 5 69520PRTArtificial SequenceHomo
sapiens glutamate decarboxylase epitope 695Phe Arg Glu Arg Gln Ser
Ser Lys Asn Leu Leu Ser Cys Glu Asn Ser 1 5 10 15 Asp Arg Asp Ala
20 69620PRTArtificial SequenceHomo sapiens Glutamate decarboxylase
1 epitope 696Met Ala Ser Ser Thr Pro Ser Ser Ser Ala Thr Ser Ser
Asn Ala Gly 1 5 10 15 Ala Asp Pro Asn 20 69719PRTArtificial
SequenceHomo sapiens Glutamate decarboxylase 2 epitope 697Pro Gly
Ser Gly Phe Trp Ser Phe Gly Ser Glu Asp Gly Ser Gly Asp 1 5 10 15
Ser Glu Asn 69815PRTArtificial SequenceHomo sapiens glutamate
receptor, ionotropic, N-methyl D-aspartate 2A epitope 698Ser Val
Ser Tyr Asp Asp Trp Asp Tyr Ser Leu Glu Ala Arg Val 1 5 10 15
69914PRTArtificial SequenceHomo sapiens glutathione peroxidase-GI
epitope 699Asn Glu His Pro Val Phe Ala Tyr Leu Lys Asp Lys Leu Pro
1 5 10 70014PRTArtificial SequenceTriticum aestivum Glutenin, high
molecular weight subunit DX5 epitope 700Ala Gln Gly Gln Gln Pro Gly
Gln Gly Gln Gln Gly Gln Gln 1 5 10 7015PRTArtificial
SequenceTriticum aestivum Glutenin, high molecular weight subunit
DX5 precursor epitope 701Gln Gln Pro Gly Gln 1 5 7025PRTArtificial
SequenceTriticum aestivum Glutenin, low molecular weight subunit
precursor epitope 702Gln Gln Gln Pro Pro 1 5 70315PRTArtificial
SequencePhaseolus vulgaris Glycine-rich cell wall structural
protein 1.8 precursor epitope 703Gly Gly Tyr Gly Asp Gly Gly Ala
His Gly Gly Gly Tyr Gly Gly 1 5 10 15 70415PRTArtificial
SequenceArachis hypogaea Glycinin epitope 704Ala Leu Ser Arg Leu
Val Leu Arg Arg Asn Ala Leu Arg Arg Pro 1 5 10 15
70513PRTArtificial SequenceGlycine max Glycinin G1 precursor
epitope 705Gly Ala Ile Val Thr Val Lys Gly Gly Leu Ser Val Ile 1 5
10 70615PRTArtificial SequenceGlycine max Glycinin G2 precursor
epitope 706Ala Leu Ser Arg Cys Thr Leu Asn Arg Asn Ala Leu Arg Arg
Pro 1 5 10 15 70715PRTArtificial SequenceHolcus lanatus group V
allergen epitope 707Ala Asn Val Pro Pro Ala Asp Lys Tyr Lys Thr Phe
Glu Ala Ala 1 5 10 15 70838PRTArtificial SequenceHomo sapiens Gu
protein epitope 708Ile Asp Ala Pro Lys Pro Lys Lys Met Lys Lys Glu
Lys Glu Met Asn 1 5 10 15 Gly Glu Thr Arg Glu Lys Ser Pro Lys Leu
Lys Asn Gly Phe Pro His 20 25 30 Pro Glu Pro Asp Cys Asn 35
70917PRTArtificial SequenceHomo sapiens H1 histone family, member 0
epitope 709Lys Glu Ile Lys Lys Val Ala Thr Pro Lys Lys Ala Ser Lys
Pro Lys 1 5 10 15 Lys 71012PRTArtificial SequenceHomo sapiens heat
shock 60kDa protein 1 (chaperonin) epitope 710Ala Tyr Ala Lys Asp
Val Lys Phe Gly Ala Asp Ala 1 5 10 7116PRTArtificial SequenceHomo
sapiens Heat shock protein HSP 90-beta epitope 711Gly Leu Glu Leu
Pro Glu 1 5 71215PRTArtificial SequenceHomo sapiens high mobility
group protein 17 epitope 712Lys Lys Ala Pro Ala Lys Lys Gly Glu Lys
Val Pro Lys Gly Lys 1 5 10 15 71322PRTArtificial SequenceHomo
sapiens High mobility group protein B1 epitope 713Ala Lys Gly Lys
Pro Asp Ala Ala Lys Lys Gly Val Val Lys Ala Glu 1 5 10 15 Lys Ser
Lys Lys Lys Lys 20 71415PRTArtificial SequenceHomo sapiens
high-mobility group box 2 epitope 714Phe Glu Asp Met Ala Lys Ser
Asp Lys Ala Arg Tyr Asp Arg Glu 1 5 10 15 71543PRTArtificial
SequenceHomo sapiens histidyl-tRNA synthetase, cytoplasmic epitope
715Ala Glu Arg Ala Ala Leu Glu Glu Leu Val Lys Leu Gln Gly Glu Arg
1 5 10 15 Val Arg Gly Leu Lys Gln Gln Lys Ala Ser Ala Glu Leu Ile
Glu Glu 20 25 30 Glu Val Ala Lys Leu Leu Lys Leu Lys Ala Gln 35 40
71616PRTArtificial SequenceHomo sapiens Histone H1.4 epitope 716Ser
Glu Thr Ala Pro Ala Ala Pro Ala Ala Pro Ala Pro Ala Glu Lys 1 5 10
15 71715PRTArtificial SequenceHomo sapiens histone H1b epitope
717Lys Pro Lys Ala Ala Lys Pro Lys Lys Ala Ala Ala Lys Lys Lys 1 5
10 15 71820PRTArtificial SequenceHomo sapiens Histone H2A.Z epitope
718Gly Lys Ala Lys Thr Lys Ala Val Ser Arg Ser Gln Arg Ala Gly Leu
1 5 10 15 Gln Phe Pro Val 20 71915PRTArtificial SequenceHomo
sapiens histone H3 epitope 719Leu Pro Phe Gln Arg Leu Val Arg Glu
Ile Ala Gln Asp Phe Lys 1 5 10 15 72010PRTArtificial SequenceHomo
sapiens Histone H3-like centromeric protein A epitope 720Lys Pro
Glu Ala Pro Arg Arg Arg Ser Pro 1 5 10 7218PRTArtificial
SequenceHomo sapiens HLA class I histocompatibility antigen, B-27
alpha chain precursor epitope 721Lys Ala Lys Ala Gln Thr Asp Arg 1
5 72216PRTArtificial SequenceHomo sapiens HLA-B27 epitope 722Ala
Lys Ala Gln Thr Asp Arg Glu Asp Leu Arg Thr Leu Leu Arg Tyr 1 5 10
15 72320PRTArtificial SequenceHomo sapiens HLA-DR3 epitope 723Arg
Pro Asp Ala Glu Tyr Trp Asn Ser Gln Lys Asp Leu Leu Glu Gln 1 5 10
15 Lys Arg Gly Arg 20 72415PRTArtificial SequenceHomo sapiens
HMG-17 epitope 724Asp Gly Lys Ala Lys Val Lys Asp Glu Pro Gln Arg
Arg Ser Ala 1 5 10 15 72513PRTArtificial SequenceHomo sapiens
HNRNPA2B1 protein epitope 725Glu Thr Thr Glu Glu Ser Leu Arg Asn
Tyr Tyr Glu Gln 1 5 10 72635PRTArtificial SequenceHomo sapiens
hypothetical protein epitope 726Ala Asn Glu Asp Ala Ala Gln Gly Ile
Ala Asn Trp Asp Ala Val Gln 1 5 10 15 Asp Ile Ala Asn Glu Asp Gly
Phe His Gly Ile Asp Ile Glu Asp Ala 20 25 30 Ala Gln Gly 35
72712PRTArtificial SequenceOryza sativa Japonica Group hypothetical
protein epitope 727Ala Phe Asn His Phe Gly Ile Gln Leu Val Gln Arg
1 5 10 72820PRTArtificial SequenceHomo sapiens Ig alpha-1 chain C
region epitope 728Pro Val Pro Ser Thr Pro Pro Thr Pro Ser Pro Ser
Thr Pro Pro Thr 1 5 10 15 Pro Ser Pro Ser 20 7297PRTArtificial
SequenceHomo sapiens Ig gamma-1 chain C region epitope 729Lys Phe
Asn Trp Tyr Val Asp 1 5 7307PRTArtificial SequenceHomo sapiens Ig
gamma-3 chain C region epitope 730Asp Gly Ser Phe Phe Leu Tyr 1 5
7317PRTArtificial SequenceHomo sapiens Ig heavy chain V-III region
(ART) epitope 731Cys Ser Val Met His Glu Gly 1 5 73216PRTArtificial
SequenceHomo sapiens Ig lambda chain V-II region MGC epitope 732Ser
Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr 1 5 10
15 7337PRTArtificial SequenceHomo sapiens Ig L-chain V-region
epitope 733Ala Pro Ser Val Thr Leu Phe 1 5 7347PRTArtificial
SequenceHomo sapiens Immunoglobulin heavy chain epitope 734Asp Lys
Ser Arg Trp Gln Glu 1 5 73516PRTArtificial SequenceHomo sapiens
immunoglobulin light chain epitope 735Lys Ala Thr Leu Val Cys Leu
Ile Ser Asp Phe Tyr Pro Gly Ala Val 1 5 10 15 7367PRTArtificial
SequenceHomo sapiens immunoglobulin light chain variable region
epitope 736Ala Gly Glu Lys Val Thr Met 1 5 7373PRTArtificial
SequenceHomo sapiens Insulin precursor epitope 737Thr Ser Ile 1
73814PRTArtificial SequenceHomo sapiens Integrin alpha-6 epitope
738Leu Lys Arg Asp Met Lys Ser Ala His Leu Leu Pro Glu His 1 5 10
73918PRTArtificial SequenceHomo sapiens Integrin beta-3 precursor
epitope 739Cys Ala Pro Glu Ser Ile Glu Phe Pro Val Ser Glu Ala Arg
Val Leu 1 5 10 15 Glu Asp 74014PRTArtificial SequenceHomo sapiens
interferon alpha 2 epitope 740Cys Asp Leu Pro Gln Thr His Ser Leu
Gly Ser Arg Arg Thr 1 5 10 74114PRTArtificial SequenceHomo sapiens
interferon alpha A epitope 741Glu Asp Ser Ile Leu Ala Val Arg Lys
Tyr Phe Gln Arg Ile 1 5 10 74212PRTArtificial SequenceHomo sapiens
interferon beta precursor epitope 742His Leu Lys Arg Tyr Tyr Gly
Arg Ile Leu His Tyr 1 5 10 74314PRTArtificial SequenceHomo sapiens
interferon-alpha 2 epitope 743Leu Met Leu Leu Ala Gln Met Arg Arg
Ile Ser Leu Phe Ser 1 5 10 74437PRTArtificial SequenceHomo sapiens
Islet amyloid polypeptide precursor epitope 744Met Gly Ile Leu Lys
Leu Gln Val Phe Leu Ile Val Leu Ser Val Ala 1 5 10 15 Leu Asn His
Leu Lys Ala Thr Pro Ile Glu Ser His Gln Val Glu Lys 20 25 30 Arg
Lys Cys Asn Thr 35 74514PRTArtificial SequenceBos taurus
Kappa-casein precursor epitope 745Ala Lys Tyr Ile Pro Ile Gln Tyr
Val Leu Ser Arg Tyr Pro 1 5 10 74610PRTArtificial SequenceHomo
sapiens Ku antigen epitope 746Arg Gly Asp Gly Pro Phe Arg Leu Gly
Gly 1 5 10 74715PRTArtificial SequenceHomo sapiens leukotriene B4
receptor 2 epitope 747Gly Arg Gly Asn Gly Asp Pro Gly Gly Gly Met
Glu Lys Asp Gly 1 5 10 15 74814PRTArtificial SequenceHomo sapiens
liver histone H1e epitope 748Ile Lys Lys Val Ala Thr Pro Lys Lys
Ala Ser Pro Lys Lys 1 5 10 74923PRTArtificial SequenceHomo sapiens
Lupus La protein epitope 749Ala Gln Pro Gly Ser Gly Lys Gly Lys Val
Gln Phe Gln Gly Lys Lys 1 5 10 15 Thr Lys Phe Ala Ser Asp Asp 20
75030PRTArtificial SequenceHomo sapiens lymphocyte activation gene
3 protein precursor epitope 750Gly Pro Pro Ala Ala Ala Pro Gly His
Pro Leu Ala Pro Gly Pro His 1 5 10 15 Pro Ala Ala Pro Ser Ser Trp
Gly Pro Arg Pro Arg Arg Tyr 20 25 30 75110PRTArtificial
SequenceHomo sapiens m3 muscarinic cholinergic receptor epitope
751Glu Pro Thr Ile Thr Phe Gly Thr Ala Ile 1 5 10
75220PRTArtificial SequenceAlternaria alternata Major allergen Alt
a 1 precursor epitope 752Ala Asp Pro Val Thr Thr Glu Gly Asp Tyr
Val Val Lys Ile Ser Glu 1 5 10 15 Phe Tyr Gly Arg 20
75315PRTArtificial SequenceAnisakis simplex Major allergen Ani s 1
epitope 753Cys Lys Met Pro Asp Arg Gly Ala Cys Ala Leu Gly Lys Lys
Pro 1 5 10 15 75413PRTArtificial SequenceAspergillus fumigatus
Major allergen Asp f 1 epitope 754Leu Asn Pro Lys Thr Asn Lys Trp
Glu Asp Lys Arg Tyr 1 5 10 75510PRTArtificial SequenceAspergillus
fumigatus Major allergen Asp f 2 epitope 755Ala His Ile Leu Arg Trp
Gly Asn Glu Ser 1 5 10 75620PRTArtificial SequenceBos taurus major
allergen beta-lactoglobulin epitope 756Leu Gln Lys Trp Glu Asn Asp
Glu Cys Ala Gln Lys Lys Ile Ile Ala 1 5 10 15 Glu Lys Thr Lys 20
75714PRTArtificial SequenceFelis catus Major allergen I polypeptide
chain 1 precursor epitope 757Asp Ala Lys Met Thr Glu Glu Asp Lys
Glu Asn Ala Leu Ser 1 5 10 75814PRTArtificial SequenceFelis catus
Major allergen I polypeptide chain 2 precursor epitope 758Glu Pro
Glu Arg Thr Ala Met Lys Lys Ile Gln Asp Cys Tyr 1 5 10
75911PRTArtificial SequenceFelis catus major allergen I,
polypeptide chain 1 epitope 759Leu Leu Asp Lys Ile Tyr Thr Ser Pro
Leu Cys 1 5 10 76029PRTArtificial SequenceTurbo cornutus major
allergen Tur c1 - Turbo cornutus epitope 760Leu Glu Asp Glu Leu Leu
Ala Glu Lys Glu Lys Tyr Lys Ala Ile Ser 1 5 10 15 Asp Glu Leu Asp
Gln Thr Phe Ala Glu Leu Ala Gly Tyr 20 25 76125PRTArtificial
SequenceDermatophagoides pteronyssinus major house dust allergen
epitope 761Leu Ala His Arg Asn Gln Ser Leu Asp Leu Ala Glu Gln Glu
Leu Val 1 5 10 15 Asp Cys Ala Ser Gln His Gly Cys His 20 25
7629PRTArtificial SequenceHevea brasiliensis Major latex allergen
Hev b 5 epitope 762Ala Pro Pro Ala Ser Glu Gln Glu Thr 1 5
76343PRTArtificial SequenceDermatophagoides pteronyssinus Major
mite fecal allergen Der p 1 epitope 763Ala Arg Glu Gln Ser Cys Arg
Arg Pro Asn Ala Gln Arg Phe Gly Ile 1 5 10 15 Ser Asn Tyr Cys Gln
Ile Tyr Pro Pro Asn Ala Asn Lys Ile Arg Glu 20 25 30 Ala Leu Ala
Gln Pro Gln Arg Tyr Cys Arg His 35 40 76412PRTArtificial
SequenceOlea europaea Major pollen allergen epitope 764Phe Thr Glu
Val Gly Tyr Thr Arg Ala Glu Gly Leu 1 5 10 76510PRTArtificial
SequenceBetula pendula Major pollen allergen Bet v 1-A epitope
765Asp Gly Asp Asn Leu Phe Pro Lys Val Ala 1 5 10
76611PRTArtificial SequenceChamaecyparis obtusa Major pollen
allergen Cha o 1 precursor epitope 766Trp Arg Ser Thr Gln Asp Ser
Phe Asn Asn Gly 1 5 10 76717PRTArtificial SequenceCorylus avellana
Major pollen allergen Cor a 1 epitope 767Tyr Val Leu Asp Gly Asp
Lys Leu Leu Pro Lys Val Ala Pro Gln Ala 1 5 10 15 Leu
76827PRTArtificial SequenceHolcus lanatus Major pollen allergen Hol
l 1 precursor epitope 768Ala Lys Ser Thr Trp Tyr Gly Lys Pro Thr
Gly Ala Gly Pro Lys Asp 1 5 10 15 Asn Gly Gly Ala Cys Gly Tyr Lys
Asp Val Asp 20 25 76912PRTArtificial SequenceJuniperus ashei Major
pollen allergen Jun a 1 precursor epitope 769Ala Phe Asn Gln Phe
Gly Pro Asn Ala Gly Gln Arg 1 5 10 77034PRTArtificial SequenceOlea
europaea major pollen allergen Ole e 1 epitope 770Ser Gly Arg Lys
Asp Cys Asn Glu Ile Pro Thr Glu Gly Trp Val Lys 1 5 10 15 Pro Ser
Leu Lys Phe Ile Leu Asn Thr Val Asn Gly Thr Thr Arg Thr 20 25 30
Val Asn 7719PRTArtificial SequenceMalus x domestica mal d 3 epitope
771Arg Thr Thr Ala Asp Arg Gln Thr Ala 1 5 77215PRTArtificial
SequenceHomo sapiens MBP protein epitope 772Glu Asn Pro Val Val His
Phe Phe Lys Asn Ile Val Thr Pro Arg 1 5 10 15 77330PRTArtificial
SequenceHomo sapiens melanin-concentrating hormone receptor 1,
isoform CRA_a epitope 773Ala Glu His Ala Ser Arg Met Ser Val Leu
Arg Ala Lys Pro Met Ser 1 5 10 15 Asn Ser Gln Arg Leu Leu Leu Leu
Ser Pro Gly Ser Pro Pro 20 25 30 77416PRTArtificial SequenceHomo
sapiens Melanocyte protein Pmel 17 precursor epitope 774Gln Val Pro
Thr Thr Glu Val Val Gly Thr Thr Pro Gly Gln Ala Pro 1 5 10 15
7756PRTArtificial SequenceHomo sapiens MHC classII HLA-DRB1 epitope
775Glu Gln Arg Arg Ala Ala 1 5 77620PRTArtificial SequenceHomo
sapiens MHC HLA-DR1-beta epitope 776Arg Pro Asp Ala Glu Tyr Trp Asn
Ser Gln Lys Asp Leu Leu Glu Gln 1 5 10 15 Arg Arg Ala Ala
20 77716PRTArtificial SequenceBlomia tropicalis Mite allergen Blo t
5 epitope 777Glu Glu Ala Gln Thr Leu Ser Lys Ile Leu Leu Lys Asp
Leu Lys Glu 1 5 10 15 7785PRTArtificial SequenceDermatophagoides
farinae Mite group 2 allergen Der f 2 precursor epitope 778Asp Pro
Cys Ile Ile 1 5 77915PRTArtificial SequenceDermatophagoides
pteronyssinus Mite group 2 allergen Der p 2 precursor epitope
779Asp Gln Val Asp Val Lys Asp Cys Ala Asn His Glu Ile Lys Lys 1 5
10 15 78033PRTArtificial SequenceLepidoglyphus destructor Mite
group 2 allergen Lep d 2 precursor epitope 780Ala Ala Asn Gln Asp
Thr Ala Lys Val Thr Ile Lys Val Leu Ala Lys 1 5 10 15 Val Ala Gly
Thr Thr Ile Gln Val Pro Gly Leu Glu Thr Asp Gly Cys 20 25 30 Lys
7816PRTArtificial SequenceTriticum aestivum monomeric alpha-amylase
inhibitor epitope 781Ala Ala Ser Val Pro Glu 1 5 78221PRTArtificial
SequenceHomo sapiens Muscarinic acetylcholine receptor M1 epitope
782Gln Tyr Leu Val Gly Glu Arg Thr Val Leu Ala Gly Gln Cys Tyr Ile
1 5 10 15 Gln Phe Leu Ser Gln 20 78317PRTArtificial SequenceHomo
sapiens myelin associated glycoprotein epitope 783Asp Ser Tyr Thr
Leu Thr Glu Glu Leu Ala Tyr Ala Glu Ile Arg Val 1 5 10 15 Lys
78413PRTArtificial SequenceHomo sapiens Myelin basic protein
epitope 784Ile Val Thr Pro Arg Thr Pro Pro Pro Ser Gln Gly Lys 1 5
10 78526PRTArtificial SequenceHomo sapiens myelin oligodendrocyte
glycoprotein epitope 785Ala Leu Val Gly Asp Glu Val Glu Leu Pro Cys
Arg Ile Ser Pro Gly 1 5 10 15 Lys Asn Ala Thr Gly Met Glu Leu Gly
Trp 20 25 7865PRTArtificial SequenceHomo sapiens myelin
oligodendrocyte glycoprotein isoform alpha6 precursor epitope
786His Arg Thr Phe Glu 1 5 7875PRTArtificial SequenceHomo sapiens
myelin proteolipid protein epitope 787Ala Asp Ala Arg Met 1 5
78821PRTArtificial SequenceHomo sapiens Myelin-associated
glycoprotein precursor epitope 788Gly His Trp Gly Ala Trp Met Pro
Ser Ser Ile Ser Ala Phe Glu Gly 1 5 10 15 Thr Cys Val Ser Ile 20
78926PRTArtificial SequenceHomo sapiens Myelin-oligodendrocyte
glycoprotein precursor epitope 789Gly Gln Phe Arg Val Ile Gly Pro
Arg His Pro Ile Arg Ala Leu Val 1 5 10 15 Gly Asp Glu Val Glu Leu
Pro Cys Arg Ile 20 25 7908PRTArtificial SequenceHomo sapiens
Myeloblastin precursor epitope 790Ala His Arg Pro Pro Ser Pro Ala 1
5 79110PRTArtificial SequenceHomo sapiens Myeloperoxidase epitope
791Gly Ser Ala Ser Pro Met Glu Leu Leu Ser 1 5 10
79220PRTArtificial SequenceHomo sapiens Myosin-11 epitope 792Ala
Leu Lys Thr Glu Leu Glu Asp Thr Leu Asp Ser Thr Ala Thr Gln 1 5 10
15 Gln Glu Leu Arg 20 7938PRTArtificial SequenceHomo sapiens
Neurofilament heavy polypeptide (NF-H) (Neurofilament triplet H
protein) (200 kDa neurofilament protein) epitope 793Ala Lys Ser Pro
Glu Lys Ala Lys 1 5 79416PRTArtificial SequenceHomo sapiens
nicotinic acetylcholine receptor alpha subunit|AChR alpha subunit
epitope 794Glu Val Asn Gln Ile Val Thr Thr Asn Val Arg Leu Lys Gln
Gln Trp 1 5 10 15 79510PRTArtificial SequenceHomo sapiens
Non-histone chromosomal protein HMG-17 epitope 795Val Lys Asp Glu
Pro Gln Arg Arg Ser Ala 1 5 10 7969PRTArtificial SequencePrunus
armeniaca Non-specific lipid-transfer protein 1 epitope 796Val Asn
Pro Asn Asn Ala Ala Ala Leu 1 5 79715PRTArtificial SequencePrunus
armeniaca Non-specific lipid-transfer protein 1 (LTP 1) (Major
allergen Pru ar 3) epitope 797Leu Ala Arg Thr Thr Pro Asp Arg Arg
Thr Ala Cys Asn Cys Leu 1 5 10 15 79818PRTArtificial SequencePrunus
domestica Non-specific lipid-transfer protein 1 (LTP 1) (Major
allergen Pru d 3) epitope 798Leu Ala Arg Thr Thr Ala Asp Arg Arg
Ala Ala Cys Asn Cys Leu Lys 1 5 10 15 Gln Leu 79915PRTArtificial
SequenceMalus x domestica Non-specific lipid-transfer protein
precursor (LTP) (Allergen Mal d 3) epitope 799Ala Asp Arg Gln Thr
Ala Cys Asn Cys Leu Lys Asn Leu Ala Gly 1 5 10 15
80010PRTArtificial SequenceHomo sapiens NR2 subunit NMDA receptor
epitope 800Asp Trp Glu Tyr Ser Val Trp Leu Ser Asn 1 5 10
8018PRTArtificial SequenceHomo sapiens nuclear autoantigen Sp-100
isoform 1 epitope 801Glu Val Phe Ile Ser Ala Pro Arg 1 5
80212PRTArtificial SequenceOlea europaea Ole e 1 protein epitope
802Glu Asp Val Pro Gln Pro Pro Val Ser Gln Phe His 1 5 10
80325PRTArtificial SequenceOlea europaea Ole e 1.0102 protein
epitope 803Glu Asp Val Pro Gln Pro Pro Val Ser Gln Phe His Ile Gln
Gly Gln 1 5 10 15 Val Tyr Cys Asp Thr Cys Arg Ala Gly 20 25
80410PRTArtificial SequenceTriticum aestivum Omega gliadin storage
protein epitope 804Gln Gln Pro Gln Gln Ser Phe Pro Gln Gln 1 5 10
8057PRTArtificial SequenceTriticum aestivum omega-5 gliadin epitope
805Gln Gln Phe His Gln Gln Gln 1 5 80635PRTArtificial
SequenceAspergillus fumigatus Oryzin precursor epitope 806Ala Ser
Asn Thr Ser Pro Ala Ser Ala Pro Asn Ala Leu Thr Val Ala 1 5 10 15
Ala Ile Asn Lys Ser Asn Ala Arg Ala Ser Phe Ser Asn Tyr Gly Ser 20
25 30 Val Val Asp 35 8079PRTArtificial SequenceGallus gallus
Ovalbumin epitope 807Cys Phe Asp Val Phe Lys Glu Leu Lys 1 5
80810PRTArtificial SequenceGallus gallus Ovomucoid epitope 808Cys
Asn Phe Cys Asn Ala Val Val Glu Ser 1 5 10 80914PRTArtificial
SequenceGallus gallus Ovomucoid precursor epitope 809Ala Glu Val
Asp Cys Ser Arg Phe Pro Asn Ala Thr Asp Lys 1 5 10
81010PRTArtificial SequenceGlycine max P34 probable thiol protease
precursor epitope 810Ala Ser Trp Asp Trp Arg Lys Lys Gly Val 1 5 10
81110PRTArtificial SequenceGlycine max P34 probable thiol protease
precursor; Gly m 1 epitope 811Pro Gln Glu Phe Ser Lys Lys Thr Tyr
Gln 1 5 10 8129PRTArtificial SequenceHomo sapiens p70 autoantigen
epitope 812Glu Ala Leu Thr Lys His Phe Gln Asp 1 5
81320PRTArtificial SequenceHomo sapiens PADI-H protein epitope
813Lys Ala Ala Ser Gly Ser Thr Gly Asp Gln Lys Val Gln Ile Ser Tyr
1 5 10 15 Tyr Gly Pro Lys 20 8149PRTArtificial SequenceParietaria
judaica Par j epitope 814Gly Thr Ser Ser Cys Arg Leu Val Pro 1 5
81547PRTArtificial SequenceBlomia tropicalis Paramyosin epitope
815Glu Lys Leu Arg Asp Gln Lys Glu Ala Leu Ala Arg Glu Asn Lys Lys
1 5 10 15 Leu Ala Asp Asp Leu Ala Glu Ala Lys Ser Gln Leu Asn Asp
Ala His 20 25 30 Arg Arg Ile His Glu Gln Glu Ile Glu Ile Lys Arg
Leu Glu Asn 35 40 45 8168PRTArtificial SequenceGadus morhua
callarias Parvalbumin beta epitope 816Ala Ala Glu Ala Ala Cys Phe
Lys 1 5 81715PRTArtificial SequenceSalmo salar parvalbumin like 1
epitope 817Ala Asp Ile Lys Thr Ala Leu Glu Ala Arg Lys Ala Ala Asp
Thr 1 5 10 15 81812PRTArtificial SequenceJuniperus ashei
Pathogenesis-related protein precursor epitope 818Ala Asp Ile Asn
Ala Val Cys Pro Ser Glu Leu Lys 1 5 10 81912PRTArtificial
SequenceNicotiana tabacum Pectate lyase epitope 819Ala Tyr Asn His
Phe Gly Lys Arg Leu Asp Gln Arg 1 5 10 82012PRTArtificial
SequenceMusa acuminata AAA Group pectate lyase 2 epitope 820Ala Phe
Asn His Phe Gly Glu Gly Leu Ile Gln Arg 1 5 10 82115PRTArtificial
SequenceFarfantepenaeus aztecus Pen a 1 allergen epitope 821Ala Asn
Ile Gln Leu Val Glu Lys Asp Lys Ala Leu Ser Asn Ala 1 5 10 15
82243PRTArtificial SequenceDermatophagoides pteronyssinus Peptidase
1 precursor (Major mite fecal allergen Der p 1) (Allergen Der p I)
epitope 822Ala Arg Glu Gln Ser Cys Arg Arg Pro Asn Ala Gln Arg Phe
Gly Ile 1 5 10 15 Ser Asn Tyr Cys Gln Ile Tyr Pro Pro Asn Val Asn
Lys Ile Arg Glu 20 25 30 Ala Leu Ala Gln Thr His Ser Ala Ile Ala
Val 35 40 8233PRTArtificial SequenceHomo sapiens pericentriolar
material 1 protein epitope 823Lys Asp Cys 1 82420PRTArtificial
SequenceHomo sapiens Periplakin epitope 824Ile His Asp Arg Lys Ser
Gly Lys Lys Phe Ser Ile Glu Glu Ala Leu 1 5 10 15 Gln Ser Gly Arg
20 82545PRTArtificial SequenceApis mellifera Phospholipase A2
precursor epitope 825Leu Ile Asp Thr Lys Cys Tyr Lys Leu Glu His
Pro Val Thr Gly Cys 1 5 10 15 Gly Glu Arg Thr Glu Gly Arg Cys Leu
His Tyr Thr Val Asp Lys Ser 20 25 30 Lys Pro Lys Val Tyr Gln Trp
Phe Asp Leu Arg Lys Tyr 35 40 45 82614PRTArtificial
SequenceMyrmecia pilosula Pilosulin-1 precursor (Major allergen Myr
p 1) (Myr p I) epitope 826Lys Glu Ala Ile Pro Met Ala Val Glu Met
Ala Lys Ser Gln 1 5 10 8278PRTArtificial SequenceHomo sapiens
plasma protease C1 inhibitor precursor epitope 827Ala Ser Ala Ile
Ser Val Ala Arg 1 5 8286PRTArtificial SequenceHomo sapiens platelet
glycoprotein IIIa epitope 828Arg Ala Arg Ala Lys Trp 1 5
82912PRTArtificial SequenceHomo sapiens plexin domain containing 1,
isoform CRA_b epitope 829Asn Cys Ser Trp Cys His Val Leu Gln Arg
Cys Ser 1 5 10 83015PRTArtificial SequenceHomo sapiens PM/Scl 100kD
nucleolar protein epitope 830Cys Ile Ala Ala Lys Lys Ile Lys Gln
Ser Val Gly Asn Lys Ser 1 5 10 15 8318PRTArtificial SequenceBetula
pendula Polcalcin Bet v 4 epitope 831Phe Gly Arg Ala Asn Arg Gly
Leu 1 5 83236PRTArtificial SequencePhleum pratense Polcalcin Phl p
7 (Calcium-binding pollen allergen Phl p 7) (P7) epitope 832Ala Asp
Asp Met Glu Arg Ile Phe Lys Arg Phe Asp Thr Asn Gly Asp 1 5 10 15
Gly Lys Ile Ser Leu Ser Glu Leu Thr Asp Ala Leu Arg Thr Leu Gly 20
25 30 Ser Thr Ser Ala 35 83325PRTArtificial SequenceLolium perenne
pollen allergen epitope 833Glu Gly Gly Thr Lys Ser Glu Val Glu Asp
Val Ile Pro Glu Gly Trp 1 5 10 15 Lys Ala Asp Thr Ser Tyr Ser Ala
Lys 20 25 83412PRTArtificial SequenceAmbrosia artemisiifolia Pollen
allergen Amb a 1.4 epitope 834Ala Phe Asn Lys Phe Thr Asp Asn Val
Asp Gln Arg 1 5 10 8358PRTArtificial SequenceAmbrosia
artemisiifolia Pollen allergen Amb a 2 precursor epitope 835Met Pro
Arg Cys Arg Phe Gly Phe 1 5 83615PRTArtificial SequenceAmbrosia
artemisiifolia var. elatior Pollen allergen Amb a 3 epitope 836Cys
Asp Ile Lys Asp Pro Ile Arg Leu Glu Pro Gly Gly Pro Asp 1 5 10 15
83731PRTArtificial SequenceBetula pendula pollen allergen Bet v 1
epitope 837Lys Ala Glu Gln Val Lys Ala Ser Lys Glu Met Gly Glu Thr
Leu Leu 1 5 10 15 Arg Ala Val Glu Ser Tyr Leu Leu Ala His Ser Asp
Ala Tyr Asn 20 25 30 83820PRTArtificial SequencePoa pratensis
Pollen allergen KBG 60 precursor epitope 838Ala Ala Asn Lys Tyr Lys
Thr Phe Val Ala Thr Phe Gly Ala Ala Ser 1 5 10 15 Asn Lys Ala Phe
20 83925PRTArtificial SequenceLolium perenne Pollen allergen Lol p
2-A (Lol p II-A) epitope 839Glu Lys Gly Met Arg Asn Val Phe Asp Asp
Val Val Pro Ala Asp Phe 1 5 10 15 Lys Val Gly Thr Thr Tyr Lys Pro
Glu 20 25 84027PRTArtificial SequenceLolium perenne Pollen allergen
Lol p 3 (Lol p III) epitope 840Lys Gly Gly Met Lys Asn Val Phe Asp
Glu Val Ile Pro Thr Ala Phe 1 5 10 15 Thr Val Gly Lys Thr Tyr Thr
Pro Glu Tyr Asn 20 25 84112PRTArtificial SequenceLolium perenne
Pollen allergen Lol p VA precursor epitope 841Ala Ala Glu Gly Ala
Thr Pro Glu Ala Lys Tyr Asp 1 5 10 84215PRTArtificial
SequencePhleum pratense Pollen allergen Phl p 1 precursor epitope
842Ala Pro Tyr His Phe Asp Leu Ser Gly His Ala Phe Gly Ala Met 1 5
10 15 8438PRTArtificial SequenceZea mays pollen allergen Phl p 11
epitope 843Arg Asp Arg Ala Arg Val Pro Leu 1 5 84412PRTArtificial
SequencePhleum pratense pollen allergen Phl pI epitope 844Ile Pro
Lys Val Pro Pro Gly Pro Asn Ile Thr Ala 1 5 10 84520PRTArtificial
SequenceCryptomeria japonica Polygalacturonase precursor epitope
845Gly Gln Cys Lys Trp Val Asn Gly Arg Glu Ile Cys Asn Asp Arg Asp
1 5 10 15 Arg Pro Thr Ala 20 84630PRTArtificial SequenceParietaria
judaica Probable non-specific lipid-transfer protein epitope 846Gln
Glu Thr Cys Gly Thr Met Val Arg Ala Leu Met Pro Cys Leu Pro 1 5 10
15 Phe Val Gln Gly Lys Glu Lys Glu Pro Ser Lys Gly Cys Cys 20 25 30
84710PRTArtificial SequenceParietaria judaica Probable non-specific
lipid-transfer protein 2 epitope 847Ala Glu Val Pro Lys Lys Cys Asp
Ile Lys 1 5 10 84830PRTArtificial SequenceParietaria judaica
Probable non-specific lipid-transfer protein 2 precursor epitope
848Glu Ala Cys Gly Lys Val Val Gln Asp Ile Met Pro Cys Leu His Phe
1 5 10 15 Val Lys Gly Glu Glu Lys Glu Pro Ser Lys Glu Cys Cys Ser
20 25 30 84912PRTArtificial SequenceSolanum lycopersicum Probable
pectate lyase P59 epitope 849Ala Phe Asn His Phe Gly Lys Arg Leu
Ile Gln Arg 1 5 10 85011PRTArtificial SequenceHomo sapiens
profilaggrin epitope 850Gly Gly Gln Gly Ser Arg His Gln Gln Ala Arg
1 5 10 85110PRTArtificial SequenceCucumis melo profilin epitope
851Ala Phe Arg Leu Glu Glu Ile Ala Ala Ile 1 5 10
85256PRTArtificial SequenceGlycine max Profilin-1 epitope 852Trp
Ala Gln Ser Thr Asp Phe Pro Gln Phe Lys Pro Glu Glu Ile Thr 1 5 10
15 Ala Ile Met Asn Asp Phe Asn Glu Pro Gly Ser Leu Ala Pro Thr Gly
20 25 30 Leu Tyr Leu Gly Gly Thr Lys Tyr Met Val Ile Gln Gly Glu
Pro Gly 35 40 45 Ala Val Ile Arg Gly Lys Lys Gly 50 55
85343PRTArtificial SequenceHevea brasiliensis Pro-hevein precursor
epitope 853Glu Gln Cys Gly Arg Gln Ala Gly Gly Lys Leu Cys Pro Asn
Asn Leu 1 5 10 15 Cys Cys Ser Gln Trp Gly Trp Cys Gly Ser Thr Asp
Glu Tyr Cys Ser 20 25 30 Pro Asp His Asn Cys Gln Ser Asn Cys Lys
Asp 35 40 85415PRTArtificial SequenceHomo sapiens Proliferating
cell nuclear antigen epitope 854Leu Lys Tyr Tyr Leu Ala Pro Lys Ile
Glu Asp Glu Glu Gly Ser 1 5 10 15 8559PRTArtificial SequenceHomo
sapiens Proline-rich transmembrane protein 2 epitope 855His Ser Glu
Ala Glu Thr Gly Pro Pro 1 5 85615PRTArtificial SequenceHomo sapiens
proteasome (prosome, macropain) activator subunit 3 (PA28 gamma;
Ki), isoform CRA_a epitope 856Leu Asp Gly Pro Thr Tyr Lys Arg Arg
Leu Asp Glu Cys Glu
Glu 1 5 10 15 85713PRTArtificial SequenceHomo sapiens protein
tyrosine phosphatase-like autoantigen epitope 857Gly Ala His Gly
Asp Thr Thr Pro Glu Tyr Gln Asp Leu 1 5 10 85820PRTArtificial
SequenceHomo sapiens protein-arginine deiminase type-4 epitope
858Ala Phe Phe Pro Asn Met Val Asn Met Leu Val Leu Gly Lys His Leu
1 5 10 15 Gly Ile Pro Lys 20 85915PRTArtificial SequenceHomo
sapiens proteinase 3 epitope 859Cys Ala Thr Arg Leu Phe Pro Asp Phe
Phe Thr Arg Val Ala Leu 1 5 10 15 86010PRTArtificial SequencePrunus
persica pru p 1 epitope 860Gly Lys Cys Gly Val Ser Ile Pro Tyr Lys
1 5 10 86115PRTArtificial SequencePrunus dulcis prunin 1 precursor
epitope 861Glu Glu Ser Gln Gln Ser Ser Gln Gln Gly Arg Gln Gln Glu
Gln 1 5 10 15 86215PRTArtificial SequencePrunus dulcis prunin 2
precursor epitope 862Asp Ser Gln Pro Gln Gln Phe Gln Gln Gln Gln
Gln Gln Gln Gln 1 5 10 15 86311PRTArtificial SequenceHesperocyparis
arizonica putative allergen Cup a 1 epitope 863Trp Arg Phe Thr Arg
Asp Ala Phe Thr Asn Gly 1 5 10 86411PRTArtificial SequenceHomo
sapiens Putative HTLV-1-related endogenous sequence (p25) epitope
864Pro Thr Arg Ala Pro Ser Gly Pro Arg Pro Pro 1 5 10
8658PRTArtificial SequenceHomo sapiens Putative small nuclear
ribonucleoprotein polypeptide E-like protein 1 epitope 865Glu Ile
His Ser Lys Thr Lys Ser 1 5 86618PRTArtificial SequenceHomo sapiens
Receptor tyrosine-protein kinase erbB-2 precursor epitope 866Pro
Glu Ser Phe Asp Gly Asp Pro Ala Ser Asn Thr Ala Pro Leu Gln 1 5 10
15 Pro Glu 86713PRTArtificial SequenceHomo sapiens Receptor-type
tyrosine-protein phosphatase-like N precursor epitope 867Lys Glu
Arg Leu Ala Ala Leu Gly Pro Glu Gly Ala His 1 5 10
86811PRTArtificial SequenceHomo sapiens recombinant IgG2 heavy
chain epitope 868Glu Pro Gln Val Val Thr Leu Pro Pro Ser Arg 1 5 10
86920PRTArtificial SequenceHomo sapiens Replication protein A 32
kDa subunit epitope 869Arg Ser Phe Gln Asn Lys Lys Ser Leu Val Ala
Phe Lys Ile Met Pro 1 5 10 15 Leu Glu Asp Met 20 87010PRTArtificial
SequenceAspergillus fumigatus Ribonuclease mitogillin precursor
epitope 870Phe Pro Thr Phe Pro Asp Gly His Asp Tyr 1 5 10
87123PRTArtificial SequenceHomo sapiens ribosomal protein L7
epitope 871Glu Leu Lys Ile Lys Arg Leu Arg Lys Lys Phe Ala Gln Lys
Met Leu 1 5 10 15 Arg Lys Ala Arg Arg Lys Leu 20 87214PRTArtificial
SequenceHomo sapiens ribosomal protein P2 epitope 872Ser Glu Glu
Ser Asp Asp Asp Met Gly Phe Gly Leu Phe Asp 1 5 10
87312PRTArtificial SequenceMangifera indica ripening-related
pectate lyase epitope 873Ala Tyr Asn His Phe Gly Glu Gly Leu Ile
Gln Arg 1 5 10 87421PRTArtificial SequenceHomo sapiens RNA binding
protein, autoantigenic (hnRNP-associated with lethal yellow homolog
(mouse)), isoform CRA_c epitope 874Gly Gly Gly Gly Gly Gly Ser Gly
Gly Gly Gly Ser Gly Gly Gly Gly 1 5 10 15 Gly Gly Gly Ser Ser 20
87521PRTArtificial SequenceHomo sapiens Ro ribonucleoprotein
epitope 875Asp Gly Tyr Val Trp Gln Val Thr Asp Met Asn Arg Leu His
Arg Phe 1 5 10 15 Leu Cys Phe Gly Ser 20 87610PRTArtificial
SequenceHevea brasiliensis Rubber elongation factor protein epitope
876Ala Glu Asp Glu Asp Asn Gln Gln Gly Gln 1 5 10
87714PRTArtificial SequenceHomo sapiens S-arrestin epitope 877Phe
Leu Gly Glu Leu Thr Ser Ser Glu Val Ala Thr Glu Val 1 5 10
87815PRTArtificial SequenceJuglans regia seed storage protein
epitope 878Asp Asp Asn Gly Leu Glu Glu Thr Ile Cys Thr Leu Arg Leu
Arg 1 5 10 15 87915PRTArtificial SequenceArachis hypogaea seed
storage protein SSP2 epitope 879Cys Gly Leu Arg Ala Pro Gln Arg Cys
Asp Leu Asp Val Glu Ser 1 5 10 15 8808PRTArtificial SequenceGallus
gallus serine (or cysteine) proteinase inhibitor, clade B
(ovalbumin), member 3 epitope 880Arg Pro Asn Ala Thr Tyr Ser Leu 1
5 8818PRTArtificial SequenceGallus gallus Serum albumin epitope
881Gln Ser Arg Ala Thr Leu Gly Ile 1 5 88217PRTArtificial
SequenceBos taurus Serum albumin precursor epitope 882Asp Asp Ser
Pro Asp Leu Pro Lys Leu Lys Pro Asp Pro Asn Thr Leu 1 5 10 15 Cys
8838PRTArtificial SequenceHomo sapiens small nuclear
ribonucleoprotein epitope 883Pro Pro Pro Gly Ile Arg Gly Pro 1 5
8848PRTArtificial SequenceHomo sapiens small nuclear
ribonucleoprotein B' epitope 884Pro Pro Pro Gly Met Arg Gly Pro 1 5
88524PRTArtificial SequenceHomo sapiens small nuclear
ribonucleoprotein D1 polypeptide epitope 885Lys Met Thr Leu Lys Asn
Arg Glu Pro Val Gln Leu Glu Thr Leu Ser 1 5 10 15 Ile Arg Gly Asn
Arg Ile Arg Tyr 20 88623PRTArtificial SequenceHomo sapiens small
nuclear ribonucleoprotein D2 isoform 1 epitope 886Gly Lys Lys Lys
Ser Lys Pro Val Asn Lys Asp Arg Tyr Ile Ser Lys 1 5 10 15 Met Phe
Leu Arg Gly Asp Ser 20 8878PRTArtificial SequenceHomo sapiens small
nuclear ribonucleoprotein F epitope 887Glu Glu Glu Glu Asp Gly Glu
Met 1 5 8888PRTArtificial SequenceHomo sapiens small nuclear
ribonucleoprotein G epitope 888Trp Ser Lys Ala His Pro Pro Glu 1 5
8899PRTArtificial SequenceHomo sapiens small nuclear
ribonucleoprotein polypeptide A epitope 889Ala Met Lys Ile Ser Phe
Ala Lys Lys 1 5 8907PRTArtificial SequenceHomo sapiens small
nuclear ribonucleoprotein polypeptide B epitope 890Pro Pro Gly Met
Arg Pro Pro 1 5 89112PRTArtificial SequenceHomo sapiens small
nuclear ribonucleoprotein polypeptide B/B' isoform B epitope 891Met
Gly Arg Gly Ala Pro Pro Pro Gly Met Met Gly 1 5 10
8927PRTArtificial SequenceHomo sapiens small nuclear
ribonucleoprotein polypeptide C, isoform CRA_b epitope 892Ala Pro
Gly Met Arg Pro Pro 1 5 89315PRTArtificial SequenceHomo sapiens
small nuclear ribonucleoprotein polypeptide D3 epitope 893Ala Ala
Arg Gly Arg Gly Arg Gly Met Gly Arg Gly Asn Ile Phe 1 5 10 15
89412PRTArtificial SequenceHomo sapiens small nuclear
ribonucleoprotein polypeptide N variant epitope 894Val Gly Arg Ala
Thr Pro Pro Pro Gly Ile Met Ala 1 5 10 89523PRTArtificial
SequenceHomo sapiens Small nuclear ribonucleoprotein Sm D1 epitope
895Gly Arg Gly Arg Gly Arg Gly Arg Gly Arg Gly Arg Gly Arg Gly Arg
1 5 10 15 Gly Arg Gly Gly Pro Arg Arg 20 8968PRTArtificial
SequenceHomo sapiens Small nuclear ribonucleoprotein Sm D2 epitope
896Glu Glu Leu Gln Lys Arg Glu Glu 1 5 8978PRTArtificial
SequenceHomo sapiens Small nuclear ribonucleoprotein-associated
proteins B and B' epitope 897Arg Gly Val Gly Gly Pro Ser Gln 1 5
89810PRTArtificial SequenceHevea brasiliensis Small rubber particle
protein epitope 898Ala Glu Glu Val Glu Glu Glu Arg Leu Lys 1 5 10
89920PRTArtificial SequenceHomo sapiens Smoothelin epitope 899Gly
Ser Thr Met Met Gln Thr Lys Thr Phe Ser Ser Ser Ser Ser Ser 1 5 10
15 Lys Lys Met Gly 20 90015PRTArtificial SequenceHomo sapiens snRNP
polypeptide B epitope 900Pro Pro Gly Met Arg Pro Pro Met Gly Pro
Met Gly Ile Pro Pro 1 5 10 15 90114PRTArtificial SequenceHomo
sapiens spectrin, alpha, non-erythrocytic 1 (alpha-fodrin), isoform
CRA_e epitope 901Phe Gln Phe Phe Gln Arg Asp Ala Glu Glu Leu Glu
Lys Trp 1 5 10 90243PRTArtificial SequenceHomo sapiens steroid
17-alpha-hydroxylase/17,20 lyase epitope 902Glu Val Pro Asp Asp Gly
Gln Leu Pro Ser Leu Glu Gly Ile Pro Lys 1 5 10 15 Val Val Phe Leu
Ile Asp Ser Phe Lys Val Lys Ile Lys Val Arg Gln 20 25 30 Ala Trp
Arg Glu Ala Gln Ala Glu Gly Ser Thr 35 40 90315PRTArtificial
SequenceHomo sapiens Sucrase-isomaltase, intestinal epitope 903Asp
Phe Thr Tyr Asp Gln Val Ala Phe Asn Gly Leu Pro Gln Phe 1 5 10 15
90412PRTArtificial SequenceCryptomeria japonica Sugi basic protein
precursor epitope 904Asp Ala Leu Thr Leu Arg Thr Ala Thr Asn Ile
Trp 1 5 10 90548PRTArtificial SequenceAspergillus fumigatus
Superoxide dismutase epitope 905Tyr Thr Leu Pro Pro Leu Pro Tyr Pro
Tyr Asp Ala Leu Gln Pro Tyr 1 5 10 15 Ile Ser Gln Gln Ile Met Glu
Leu His His Lys Lys His His Gln Thr 20 25 30 Tyr Val Asn Gly Leu
Asn Ala Ala Leu Glu Ala Gln Lys Lys Ala Ala 35 40 45
90611PRTArtificial SequenceHomo sapiens T cell receptor beta
variable 20 epitope 906Arg Ser Leu Asp Phe Gln Ala Thr Thr Met Phe
1 5 10 90718PRTArtificial SequenceHomo sapiens T cell receptor beta
variable 5 epitope 907Ala Leu Gly Gln Gly Pro Gln Phe Ile Phe Gln
Tyr Tyr Glu Glu Glu 1 5 10 15 Glu Arg 90815PRTArtificial
SequenceHomo sapiens Tax1-binding protein 1 epitope 908Glu Phe Lys
Lys Arg Phe Ser Asp Ala Thr Ser Lys Ala His Gln 1 5 10 15
90916PRTArtificial SequenceHomo sapiens T-cell receptor beta chain
epitope 909Gln Pro Leu Lys Glu Gln Pro Ala Leu Asn Asp Ser Arg Tyr
Cys Leu 1 5 10 15 91016PRTArtificial SequenceHomo sapiens T-cell
receptor beta chain C region epitope 910Ser Ala Thr Phe Trp Gln Asn
Pro Arg Asn His Phe Arg Cys Gln Val 1 5 10 15 91116PRTArtificial
SequenceHomo sapiens T-cell receptor beta chain V region YT35
epitope 911Cys Lys Pro Ile Ser Gly His Asn Ser Leu Phe Trp Tyr Arg
Gln Thr 1 5 10 15 91210PRTArtificial SequenceHomo sapiens T-cell
receptor beta-chain (V1-D-J-C) precursor epitope 912Ser Pro Arg Ser
Gly Asp Leu Ser Val Tyr 1 5 10 91321PRTArtificial SequenceHomo
sapiens TCR V-beta 6.1 epitope 913Leu Gly Gln Gly Pro Glu Phe Leu
Ile Tyr Phe Gln Gly Thr Gly Ala 1 5 10 15 Ala Asp Asp Ser Gly 20
91410PRTArtificial SequenceHomo sapiens TCR V-beta 6.3 epitope
914Asp Pro Ile Ser Gly His Val Ser Leu Phe 1 5 10
91520PRTArtificial SequenceHomo sapiens Thyroglobulin epitope
915Pro Pro Ala Arg Ala Leu Lys Arg Ser Leu Trp Val Glu Val Asp Leu
1 5 10 15 Leu Ile Gly Ser 20 91619PRTArtificial SequenceHomo
sapiens Thyroid peroxidase epitope 916Gly Leu Pro Arg Leu Glu Thr
Pro Ala Asp Leu Ser Thr Ala Ile Ala 1 5 10 15 Ser Arg Ser
91715PRTArtificial SequenceHomo sapiens thyroid stimulating hormone
receptor epitope 917Glu Ile Ile Gly Phe Gly Gln Glu Leu Lys Asn Pro
Gln Glu Glu 1 5 10 15 91816PRTArtificial SequenceHomo sapiens
thyroid stimulating hormone receptor variant epitope 918Glu Glu Gln
Glu Asp Glu Ile Ile Gly Phe Gly Gln Glu Leu Lys Asn 1 5 10 15
91920PRTArtificial SequenceHomo sapiens Thyrotropin receptor
epitope 919Gly Gln Glu Leu Lys Asn Pro Gln Glu Glu Thr Leu Gln Ala
Phe Asp 1 5 10 15 Ser His Tyr Asp 20 92015PRTArtificial
SequenceHomo sapiens transaldolase 1 epitope 920Ala Ala Ala Gln Met
Pro Ala Tyr Gln Glu Leu Val Glu Glu Ala 1 5 10 15
92120PRTArtificial SequenceTrichophyton rubrum Tri r 2 allergen
epitope 921Asp Cys Asn Gly His Gly Thr His Val Ala Gly Thr Val Gly
Gly Thr 1 5 10 15 Lys Tyr Gly Leu 20 92215PRTArtificial
SequenceHomo sapiens trinucleotide repeat containing 6A, isoform
CRA_b epitope 922Ala Phe Leu Ser Val Asp His Leu Gly Gly Gly Gly
Glu Ser Met 1 5 10 15 92315PRTArtificial SequenceHomo sapiens
trinucleotide repeat containing 6A, isoform CRA_c epitope 923Trp
Gly Ser Ser Ser Val Gly Pro Gln Ala Leu Ser Lys Ser Gly 1 5 10 15
92436PRTArtificial SequenceHomo sapiens tripartite motif-containing
67 epitope 924Leu Gly Gly Gly Ala Gly Gly Gly Gly Asp His Ala Asp
Lys Leu Ser 1 5 10 15 Leu Tyr Ser Glu Thr Asp Ser Gly Tyr Gly Ser
Tyr Thr Pro Ser Leu 20 25 30 Lys Ser Pro Asn 35 92510PRTArtificial
SequenceTriticum aestivum Triticum aestivum proteins epitope 925Leu
Pro Gln Gln Gln Ile Pro Gln Gln Pro 1 5 10 9269PRTArtificial
SequencePenaeus tropomyosin epitope 926Phe Leu Ala Glu Glu Ala Asp
Arg Lys 1 5 92720PRTArtificial SequenceHomo sapiens TSHR protein
epitope 927Cys His Gln Glu Glu Asp Phe Arg Val Thr Cys Lys Asp Ile
Gln Arg 1 5 10 15 Ile Pro Ser Leu 20 92814PRTArtificial
SequenceHomo sapiens tubulin beta-6 chain epitope 928Ala Ala Cys
Asp Pro Arg His Gly Arg Tyr Leu Thr Val Ala 1 5 10
92910PRTArtificial SequenceHomo sapiens tumor necrosis factor
ligand superfamily member 6 epitope 929Glu Trp Glu Asp Thr Tyr Gly
Ile Val Leu 1 5 10 93020PRTArtificial SequenceParalichthys
olivaceus type 1 collagen alpha 2 epitope 930Met Lys Gly Leu Arg
Gly His Pro Gly Leu Gln Gly Met Pro Gly Pro 1 5 10 15 Ser Gly Pro
Ser 20 93110PRTArtificial SequenceTriticum aestivum type 1
non-specific lipid transfer protein precursor epitope 931Ala Arg
Gly Thr Pro Leu Lys Cys Gly Val 1 5 10 9325PRTArtificial
SequenceHomo sapiens U1 small nuclear ribonucleoprotein 70 kDa
epitope 932Glu Arg Lys Arg Arg 1 5 93326PRTArtificial SequenceHomo
sapiens U1 small nuclear ribonucleoprotein A epitope 933Ala Gly Ala
Ala Arg Asp Ala Leu Gln Gly Phe Lys Ile Thr Gln Asn 1 5 10 15 Asn
Ala Met Lys Ile Ser Phe Ala Lys Lys 20 25 9348PRTArtificial
SequenceHomo sapiens U1 small nuclear ribonucleoprotein C epitope
934Pro Ala Pro Gly Met Arg Pro Pro 1 5 93518PRTArtificial
SequenceAnisakis simplex UA3-recognized allergen epitope 935Met Cys
Gln Cys Val Gln Lys Tyr Gly Thr Glu Phe Cys Lys Lys Arg 1 5 10 15
Leu Ala 9369PRTArtificial SequenceHomo sapiens unnamed protein
product epitope 936Ala Phe Gln Gln Gly Lys Ile Pro Pro 1 5
9378PRTArtificial SequenceJuglans nigra vicilin seed storage
protein epitope 937Ser Phe Glu Asp Gln Gly Arg Arg 1 5
93815PRTArtificial SequenceAnacardium occidentale Vicilin-like
protein epitope 938Ala Ile Met Gly Pro Pro Thr Lys Phe Ser Phe Ser
Leu Phe Leu 1 5 10 15 93910PRTArtificial SequenceJuglans regia
vicilin-like protein precursor epitope 939Asp Gln Arg Ser Gln Glu
Glu Arg Glu Arg 1 5 10 94020PRTArtificial SequenceHomo sapiens
Vimentin epitope 940Arg Leu Arg Ser Ser Val Pro Gly Val Arg Leu Leu
Gln Asp Ser Val 1 5 10 15 Asp Phe Ser Leu 20 94115PRTArtificial
SequenceHomo sapiens von Willebrand factor epitope 941His Cys Gln
Ile Cys His Cys Asp Val Val Asn Leu Thr Cys Glu 1 5 10
15 94245PRTArtificial SequenceHomo sapiens von Willebrand
factor-cleaving protease precursor epitope 942Pro Ser His Phe Gln
Gln Ser Cys Leu Gln Ala Leu Glu Pro Gln Ala 1 5 10 15 Val Ser Ser
Tyr Leu Ser Pro Gly Ala Pro Leu Lys Gly Arg Pro Pro 20 25 30 Ser
Pro Gly Phe Gln Arg Gln Arg Gln Arg Gln Arg Arg 35 40 45
94315PRTArtificial SequenceHomo sapiens XRCC4 protein epitope
943Val Ser Lys Asp Asp Ser Ile Ile Ser Ser Leu Asp Val Thr Asp 1 5
10 15
* * * * *