Method of epitope discovery
A method of epitope discovery comprising the step of selecting an epitope from a population of peptide fragments of an antigen associated with a target cell, wherein the fragments have a known or predicted affinity for a major histocompatibility complex class I receptor peptide binding cleft, wherein the epitope selected corresponds to a proteasome cleavage product of the target cell.
1 . A method for selecting and/or producing a T cell epitope, comprising contacting a peptide or polypeptide to a proteasome to permit processing of the peptide or polypeptide by the proteasome to determine the c-terminus of said T cell epitope.
2 . The method of claim 1 , wherein said proteasome is an immune proteasome.
3 . The method of claim 1 , wherein said proteasome is a housekeeping proteasome.
4 . The method of claim 1 , wherein said proteasome is a proteasome exposed to interferon-gamma.
5 . The method of claim 1 , wherein said peptide or polypeptide is synthetic.
6 . The method of claim 1 , wherein said peptide or polypeptide is a fragment of an antigen.
7 . The method of claim 1 , wherein said peptide or polypeptide is an epitope cluster.
8 . The method of claim 1 , further comprising identifying additional characteristics of said T cell epitope.
9 . The method of claim 8 , wherein said characteristics comprises at least one characteristic selected from the group consisting of MHC binding affinity, identity of a c-terminus, N-terminus, and proteolytic cleavage sites.
10 . A T cell epitope obtainable by a method selected from the group consisting of the method of claim 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 and 9 .
11 . A nucleic acid encoding at least one T-cell epitope according to claim 10 .
12 . The nucleic acid of claim 11 , comprising at least two sequences, each encoding a T cell epitope, wherein said at least two sequences are separated by at least one proteolytic cleavage site.
13 . A vector comprising a nucleic acid selected from the group consisting of the nucleic acid of claim 11 and claim 12 .
14 . A composition comprising a vector of claim 13 .
15 . A composition comprising a T cell epitope of claim 10 .
16 . A polypeptide obtainable by expressing a nucleic acid of claim 11 or claim 12 .
17 . A composition comprising the polypeptide of claim 16 .
18 . A composition comprising an adjuvant and a composition selected from the group consisting of the composition of claim 14 , 15 and 17 the polypeptide of claim 16 .
19 . A method for selecting and/or producing a T cell epitope, comprising subjecting a precursor peptide or polypeptide to the action of a 20S proteasome, in particular an IFN-.gamma. or other immune-related proteasome or a functional equivalent thereof to determine the location of the c-terminus of said T cell epitope.
20 . A method for selecting and/or producing according to claim 1 wherein said precursor peptide or polypeptide is tested for other T cell epitope characteristics.
21 . A method according to claim 20 , wherein said further testing comprises testing for the right anchor residues, the proper cleavage sites, the proper pathway signals, and/or the stability of the MHC-T cell epitope complex.
22 . A T cell epitope obtainable by a method according to any one of claims 19 - 21 .
23 . A nucleic acid encoding at least one T cell epitope according to claim 22 .
24 . A nucleic acid according to claim 23 comprising at least two sequences encoding a T cell epitope, separated by at least one proteolytic cleavage site.
25 . A gene delivery vehicle comprising a nucleic acid according to claim 22 or 23 .
26 . A pharmaceutical composition comprising a gene delivery vehicle according to claim 7 .
27 . A pharmaceutical composition comprising at least one epitope according to claim 22 .
28 . A proteinaceous molecule obtainable by expression of a nucleic acid according to claim 23 or 24 .
29 . A pharmaceutical composition comprising a proteinaceous molecule according to claim 28 .
30 . A pharmaceutical composition according to claim 26 , 27 or 29 further comprising an adjuvant.
31 . A pharmaceutical composition according to claim 26 , 27 , 29 or 30 comprising an antigen presenting moiety.
32 . A pharmaceutical composition according to claim 31 wherein said moiety comprises a major histocompatibility molecule.
33 . A pharmaceutical composition according to claim 32 wherein said major histocompatibility molecule is present on an antigen presenting cell, such as a dendritic cell.
34 . A method of epitope discovery comprising the step of selecting an epitope from a population of peptide fragments of an antigen associated with a target cell, wherein the fragments have a known or predicted affinity for a major histocompatibility complex class I receptor peptide binding cleft, wherein the epitope selected corresponds to a proteasome cleavage product of the target cell.