Expression vectors encoding epitopes of target-associated antigens and methods for their design

Abstract

The invention disclosed herein is directed to methods of identifying a polypeptide suitable for epitope liberation including, for example, the steps of identifying an epitope of interest; providing a substrate polypeptide sequence including the epitope, wherein the substrate polypeptide permits processing by a proteasome; contacting the substrate polypeptide with a composition including the proteasome, under conditions that support processing of the substrate polypeptide by the proteasome; and assaying for liberation of the epitope. The invention further relates to vectors including a housekeeping epitope expression cassette. The housekeeping epitope(s) can be derived from a target-associated antigen, and the housekeeping epitope can be liberatable, that is capable of liberation, from a translation product of the cassette by immunoproteasome processing. The invention also relates to a method of activating a T cell comprising contacting a substrate polypeptide with an APC and contacting the APC with a T cell.

Description

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The invention disclosed herein is directed to methods for the design of epitope-encoding vectors for use in compositions, including for example, pharmaceutical compositions capable of inducing an immune response in a subject to whom the compositions are administered. The invention is further directed to the vectors themselves. The epitope(s) expressed using such vectors can stimulate a cellular immune response against a target cell displaying the epitope(s).

[0004] 2. Description of the Related Art

[0005] The immune system can be categorized into two discrete effector arms. The first is innate immunity, which involves numerous cellular components and soluble factors that respond to all infectious challenges. The other is the adaptive immune response, which is customized to respond specifically to precise epitopes from infectious agents. The adaptive immune response is further broken down into two effector arms known as the humoral and cellular immune systems. The humoral arm is centered on the production of antibodies by B-lymphocytes while the cellular arm involves the killer cell activity of cytotoxic T Lymphocytes.

[0006] Cytotoxic T Lymphocytes (CTL) do not recognize epitopes on the infectious agents themselves. Rather, CTL detect fragments of antigens derived from infectious agents that are displayed on the surface of infected cells. As a result antigens are visible to CTL only after they have been processed by the infected cell and thus displayed on the surface of the cell.

[0007] The antigen processing and display system on the surface of cells has been well established. CTL recognize short peptide antigens, which are displayed on the surface in non-covalent association with class I major histocompatibility complex molecules (MHC). These class I peptides are in turn derived from the degradation of cytosolic proteins.

SUMMARY OF THE INVENTION

[0008] Embodiments of the invention provide expression cassettes, for example, for use in vaccine vectors, which encode one or more embedded housekeeping epitopes, and methods for designing and testing such expression cassettes. Housekeeping epitopes can be liberated from the translation product of such cassettes through proteolytic processing by the immunoproteasome of professional antigen presenting cells (pAPC). In one embodiment of the invention, sequences flanking the housekeeping epitope(s) can be altered to promote cleavage by the immunoproteasome at the desired location(s). Housekeeping epitopes, their uses, and identification are described in U.S. patent application Ser. Nos. 09/560,465 and 09/561,074 entitled EPITOPE SYNCHRONIZATION IN ANTIGEN PRESENTING CELLS, and METHOD OF EPITOPE DISCOVERY, respectively; both of which were filed on Apr. 28, 2000, and which are both incorporated herein by reference in their entireties.

[0009] Examples of housekeeping epitopes are disclosed in provisional U.S. patent applications entitled EPITOPE SEQUENCES, Ser. Nos. 60/282,211, filed on Apr. 6, 2001; 60/337,017, filed on Nov. 7, 2001; 60/363210 filed Mar. 7, 2002; and 60/409,123, filed on Sep. 5, 2002; and U.S. application Ser. No. 10/117,937, filed on Apr. 4, 2002, which is also entitled EPITOPE SEQUENCES; which are all incorporated herein by reference in their entirety.

[0010] In other embodiments of the invention, the housekeeping epitope(s) can be flanked by arbitrary sequences or by sequences incorporating residues known to be favored in immunoproteasome cleavage sites. As used herein the term “arbitrary sequences” refers to sequences chosen without reference to the native sequence context of the epitope, their ability to promote processing, or immunological function. In further embodiments of the invention multiple epitopes can be arrayed head-to-tail. These arrays can be made up entirely of housekeeping epitopes. Likewise, the arrays can include alternating housekeeping and immune epitopes. Alternatively, the arrays can include housekeeping epitopes flanked by immune epitopes, whether complete or distally truncated. Further, the arrays can be of any other similar arrangement. There is no restriction on placing a housekeeping epitope at the terminal positions of the array. The vectors can additionally contain authentic protein coding sequences or segments thereof containing epitope clusters as a source of immune epitopes. The term “authentic” refers to natural protein sequences.

[0011] Epitope clusters and their uses are described in U.S. patent application Ser. Nos. 09/561,571 entitled EPITOPE CLUSTERS, filed on Apr. 28, 2000; 10/005,905, entitled EPITOPE SYNCHRONIZATION IN ANTIGEN PRESENTING CELLS, filed on Nov. 7, 2001; and 10/026,066, filed on Dec. 7, 2001, also entitled EPITOPE SYNCHRONIZATION IN ANTIGEN PRESENTING CELLS; all of which are incorporated herein by reference in their entirety.

[0012] Embodiments of the invention can encompass screening the constructs to determine whether the housekeeping epitope is liberated. In constructs containing multiple housekeeping epitopes, embodiments can include screening to determine which epitopes are liberated. In a preferred embodiment, a vector containing an embedded epitope can be used to immunize HLA transgenic mice and the resultant CTL can be tested for their ability to recognize target cells presenting the mature epitope. In another embodiment, target cells expressing immunoproteasome can be transformed with the vector. The target cell may express immunoproteasome either constitutively, because of treatment with interferon (IFN), or through genetic manipulation, for example. CTL that recognize the mature epitope can be tested for their ability to recognize these target cells. In yet another embodiment, the embedded epitope can be prepared as a synthetic peptide. The synthetic peptide then can be subjected to digestion by an immunoproteasome preparation in vitro and the resultant fragments can be analyzed to determine the sites of cleavage. Such polypeptides, recombinant or synthetic, from which embedded epitopes can be successfully liberated, can also be incorporated into immunogenic compositions.

[0013] The invention disclosed herein relates to the identification of a polypeptide suitable for epitope liberation. One embodiment of the invention, relates to a method of identifying a polypeptide suitable for epitope liberation including, for example, the steps of identifying an epitope of interest; providing a substrate polypeptide sequence including the epitope, wherein the substrate polypeptide permits processing by a proteasome; contacting the substrate polypeptide with a composition including the proteasome, under conditions that support processing of the substrate polypeptide by the proteasome; and assaying for liberation of the epitope.

[0014] The epitope can be embedded in the substrate polypeptide, and in some aspects the substrate polypeptide can include more than one epitope, for example. Also, the epitope can be a housekeeping epitope.

[0015] In one aspect, the substrate polypeptide can be a synthetic peptide. Optionally, the substrate polypeptide can be included in a formulation promoting protein transfer. Alternatively, the substrate polypeptide can be a fusion protein. The fusion protein can further include a protein domain possessing protein transfer activity. Further, the contacting step can include immunization with the substrate polypeptide.

[0016] In another aspect, the substrate polypeptide can be encoded by a polynucleotide. The contacting step can include immunization with a vector including the polynucleotide, for example. The immunization can be carried out in an HLA-transgenic mouse or any other suitable animal, for example. Alternatively, the contacting step can include transforming a cell with a vector including the polynucleotide. In some embodiments the transformed cell can be a target cell that is targeted by CTL for purposes of assaying for proper liberation of epitope.

[0017] The proteasome processing can take place intracellularly, either in vitro or in vivo. Further, the proteasome processing can take place in a cell-free system.

[0018] The assaying step can include a technique selected from the group including, but not limited to, mass spectrometry, N-terminal pool sequencing, HPLC, and the like. Also, the assaying step can include a T cell target recognition assay. The T cell target recognition assay can be selected from the group including, but not limited to, a cytolytic activity assay, a chromium release assay, a cytokine assay, an ELISPOT assay, tetramer analysis, and the like.

[0019] In still another aspect, the amino acid sequence of the substrate polypeptide including the epitope can be arbitrary. Also, the substrate polypeptide in which the epitope is embedded can be derived from an authentic sequence of a target-associated antigen. Further, the substrate polypeptide in which the epitope is embedded can be conformed to a preferred immune proteasome cleavage site flanking sequence.

[0020] In another aspect, the substrate polypeptide can include an array of additional epitopes. Members of the array can be arranged head-to-tail, for example. The array can include more than one housekeeping epitope. The more than one housekeeping epitope can include copies of the same epitope. The array can include a housekeeping and an immune epitope, or alternating housekeeping and immune epitopes, for example. Also, the array can include a housekeeping epitope positioned between two immune epitopes in an epitope battery. The array can include multiple epitope batteries, so that there are two immune epitopes between each housekeeping epitope in the interior of the array. Optionally, at least one of the epitopes can be truncated distally to its junction with an adjacent epitope. The truncated epitopes can be immune epitopes, for example. The truncated epitopes can have lengths selected from the group including, but not limited to, 9, 8, 7, 6, 5, 4 amino acids, and the like.

[0021] In still another aspect, the substrate polypeptide can include an array of epitopes and epitope clusters. Members of the array can be arranged head-to-tail, for example.

[0022] In yet another aspect, the proteasome can be an immune proteasome.

[0023] Another embodiment of the disclosed invention relates to vectors including a housekeeping epitope expression cassette. The housekeeping epitope(s) can be derived from a target-associated antigen, and the housekeeping epitope can be liberatable, that is capable of liberation, from a translation product of the cassette by immunoproteasome processing.

[0024] In one aspect of the invention the expression cassette can encode an array of two or more epitopes or at least one epitope and at least one epitope cluster. The members of the array can be arranged head-to-tail, for example. Also, the members of the array can be arranged head-to-tail separated by spacing sequences, for example. Further, the array can include a plurality of housekeeping epitopes. The plurality of housekeeping epitopes can include more than one copy of the same epitope or single copies of distinct epitopes, for example. The array can include at least one housekeeping epitope and at least one immune epitope. Also, the array can include alternating housekeeping and immune epitopes. Further, the array includes a housekeeping epitope sandwiched between two immune epitopes so that there are two immune epitopes between each housekeeping epitope in the interior of the array. The immune epitopes can be truncated distally to their junction with the adjacent housekeeping epitope.

[0025] In another aspect, the expression cassette further encodes an authentic protein sequence, or segment thereof, including at least one immune epitope. Optionally, the segment can include at least one epitope cluster. The housekeeping epitope expression cassette and the authentic sequence including at least one immune epitope can be encoded in a single reading frame or transcribed as a single mRNA species, for example. Also, the housekeeping epitope expression cassette and the authentic sequence including at least one immune epitope may not be transcribed as a single mRNA species.

[0026] In yet another aspect, the vector can include a DNA molecule or an RNA molecule. The vector can encode, for example, SEQ ID NO. 4, SEQ ID NO. 17, SEQ ID NO. 20, SEQ ID NO. 26, SEQ ID NO. 27, SEQ ID NO. 29, SEQ ID NO. 33, and the like. Also, the vector can include SEQ ID NO. 9, SEQ ID NO. 19, SEQ ID NO. 21, SEQ ID NO. 30, SEQ ID NO. 34, and the like. Also, the vector can encode SEQ ID NO. 5 or SEQ ID NO. 18, for example.

[0027] In still another aspect, the target-associated antigen can be an antigen derived from or associated with a tumor or an intracellular parasite, and the intracellular parasite can be, for example, a virus, a bacterium, a protozoan, or the like.

[0028] Another embodiment of the invention relates to vectors including a housekeeping epitope identified according to any of the methods disclosed herein, claimed or otherwise. For example, embodiments can relate to vector encoding a substrate polypeptide that includes a housekeeping epitope by any of the methods described herein.

[0029] In one aspect, the housekeeping epitope can be liberated from the cassette translation product by immune proteasome processing

[0030] Another embodiment of the disclosed invention relates to methods of activating a T cell. The methods can include, for example, the steps of contacting a vector including a housekeeping epitope expression cassette with an APC. The housekeeping epitope can be derived from a target-associated antigen, for example, and the housekeeping epitope can be liberatable from a translation product of the cassette by immunoproteasome processing. The methods can further include contacting the APC with a T cell. The contacting of the vector with the APC can occur in vitro or in vivo.

[0031] Another embodiment of the disclosed invention relates to a substrate polypeptide including a housekeeping epitope wherein the housekeeping epitope can be liberated by immunoproteasome processing in a pAPC.

[0032] Another embodiment of the disclosed invention relates to a method of activating a T cell comprising contacting a substrate polypeptide including a housekeeping epitope with an APC wherein the housekeeping epitope can be liberated by immunoproteasome processing and contacting the APC with a T cell.

BRIEF DESCRIPTION OF THE DRAWINGS

[0033] FIG. 1. An illustrative drawing depicting pMA2M.

[0034] FIG. 2. Assay results showing the % of specific lysis of ELAGIGILTV pulsed and unpulsed T2 target cells by mock immunized CTL.

[0035] FIG. 3. Assay results showing the % of specific lysis of ELAGIGILTV pulsed and unpulsed T2 target cells by pVAXM3 immunized CTL.

[0036] FIG. 4. Assay results showing the % of specific lysis of ELAGIGILTV pulsed and unpulsed T2 target cells by pVAXM2 immunized CTL.

[0037] FIG. 5. Assay results showing the % of specific lysis of ELAGIGILTV pulsed and unpulsed T2 target cells by pVAXM1 immunized CTL.

[0038] FIG. 6. Illustrates a sequence of SEQ ID NO. 22 from which the NY-ESO-1157-165 epitope is liberated by immunoproteasomal processing.

[0039] FIG. 7. Shows the differential processing by immunoproteasome and housekeeping proteasome of the SLLMWITQC epitope (SEQ ID NO. 12) in its native context where the cleavage following the C is more efficiently produced by housekeeping than immunoproteasome.

[0040] FIGS. 8. 8A: Shows the results of the human immunoproteasome digest of SEQ ID NO. 31. 8B: Shows the comparative results of mouse versus human immunoproteasome digestion of SEQ ID NO. 31.

[0041] FIG. 9. Shows the differential processing of SSX-231-68 by housekeeping and immunoproteasome.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0042] Definitions

[0043] Unless otherwise clear from the context of the use of a term herein, the following listed terms shall generally have the indicated meanings for purposes of this description.

[0044] PROFESSIONAL ANTIGEN-PRESENTING CELL (pAPC)—a cell that possesses T cell costimulatory molecules and is able to induce a T cell response. Well characterized pAPCs include dendritic cells, B cells, and macrophages.

[0045] PERIPHERAL CELL—a cell that is not a pAPC.

[0046] HOUSEKEEPING PROTEASOME—a proteasome normally active in peripheral cells, and generally not present or not strongly active in pAPCs.

[0047] IMMUNOPROTEASOME—a proteasome normally active in pAPCs; the immunoproteasome is also active in some peripheral cells in infected tissues or following exposure to interferon.

[0048] EPITOPE—a molecule or substance capable of stimulating an immune response. In preferred embodiments, epitopes according to this definition include but are not necessarily limited to a polypeptide and a nucleic acid encoding a polypeptide, wherein the polypeptide is capable of stimulating an immune response. In other preferred embodiments, epitopes according to this definition include but are not necessarily limited to peptides presented on the surface of cells, the peptides being non-covalently bound to the binding cleft of class I MHC, such that they can interact with T cell receptors (TCR). Epitopes presented by class I MHC may be in immature or mature form. “Mature” refers to an MHC epitope in distinction to any precursor (“immature”) that may include or consist essentially of a housekeeping epitope, but also includes other sequences in a primary translation product that are removed by processing, including without limitation, alone or in any combination, proteasomal digestion, N-terminal trimming, or the action of exogenous enzymatic activities. Thus, a mature epitope may be provided embedded in a somewhat longer polypeptide, the immunological potential of which is due, at least in part, to the embedded epitope; or in its ultimate form that can bind in the MHC binding cleft to be recognized by TCR, respectively.

[0049] MHC EPITOPE—a polypeptide having a known or predicted binding affinity for a mammalian class I or class II major histocompatibility complex (MHC) molecule.

[0050] HOUSEKEEPING EPITOPE—In a preferred embodiment, a housekeeping epitope is defined as a polypeptide fragment that is an MHC epitope, and that is displayed on a cell in which housekeeping proteasomes are predominantly active. In another preferred embodiment, a housekeeping epitope is defined as a polypeptide containing a housekeeping epitope according to the foregoing definition, that is flanked by one to several additional amino acids. In another preferred embodiment, a housekeeping epitope is defined as a nucleic acid that encodes a housekeeping epitope according to the foregoing definitions. Exemplary housekeeping epitopes are provide in U.S. application Ser. No. 10/117,937, filed on Apr. 4, 2002; and U.S. Provisional Application Nos. 60/282,211, filed on Apr. 6, 2001; 60/337,017, filed on Nov. 7, 2001; 60/363210 filed Mar. 7, 2002; and 60/409,123, filed on Sep. 5, 2002; all of which are entitled EPITOPE SEQUENCES, and all of which above were incorporated herein by reference in their entireties.

[0051] IMMUNE EPITOPE—In a preferred embodiment, an immune epitope is defined as a polypeptide fragment that is an MHC epitope, and that is displayed on a cell in which immunoproteasomes are predominantly active. In another preferred embodiment, an immune epitope is defined as a polypeptide containing an immune epitope according to the foregoing definition, that is flanked by one to several additional amino acids. In another preferred embodiment, an immune epitope is defined as a polypeptide including an epitope cluster sequence, having at least two polypeptide sequences having a known or predicted affinity for a class I MHC. In yet another preferred embodiment, an immune epitope is defined as a nucleic acid that encodes an immune epitope according to any of the foregoing definitions.

[0052] TARGET CELL—a cell to be targeted by the vaccines and methods of the invention. Examples of target cells according to this definition include but are not necessarily limited to: a neoplastic cell and a cell harboring an intracellular parasite, such as, for example, a virus, a bacterium, or a protozoan. Target cells can also include cells that are targeted by CTL as a part of assays to determine or confirm proper epitope liberation and processing by a cell expressing immunoproteasome, to determine T cell specificity or immunogenicity for a desired epitope. Such cells may be transfored to express the substrate or liberation sequence, or the cells can simply be pulsed with peptide/epitope.

[0053] TARGET-ASSOCIATED ANTIGEN (TAA)—a protein or polypeptide present in a target cell. TUMOR-ASSOCIATED ANTIGENS (TuAA)—a TAA, wherein the target cell is a neoplastic cell.

[0054] HLA EPITOPE—a polypeptide having a known or predicted binding affinity for a human class I or class II HLA complex molecule.

[0055] ANTIBODY—a natural immunoglobulin (Ig), poly- or monoclonal, or any molecule composed in whole or in part of an Ig binding domain, whether derived biochemically or by use of recombinant DNA. Examples include inter alia, F(ab), single chain Fv, and Ig variable region-phage coat protein fusions.

[0056] ENCODE—an open-ended term such that a nucleic acid encoding a particular amino acid sequence can consist of codons specifying that (poly)peptide, but can also comprise additional sequences either translatable, or for the control of transcription, translation, or replication, or to facilitate manipulation of some host nucleic acid construct.

[0057] SUBSTANTIAL SIMILARITY—this term is used to refer to sequences that differ from a reference sequence in an inconsequential way as judged by examination of the sequence. Nucleic acid sequences encoding the same amino acid sequence are substantially similar despite differences in degenerate positions or modest differences in length or composition of any non-coding regions. Amino acid sequences differing only by conservative substitution or minor length variations are substantially similar. Additionally, amino acid sequences comprising housekeeping epitopes that differ in the number of N-terminal flanking residues, or immune epitopes and epitope clusters that differ in the number of flanking residues at either terminus, are substantially similar. Nucleic acids that encode substantially similar amino acid sequences are themselves also substantially similar.

[0058] FUNCTIONAL SIMILARITY—this term is used to refer to sequences that differ from a reference sequence in an inconsequential way as judged by examination of a biological or biochemical property, although the sequences may not be substantially similar. For example, two nucleic acids can be useful as hybridization probes for the same sequence but encode differing amino acid sequences. Two peptides that induce cross-reactive CTL responses are functionally similar even if they differ by non-conservative amino acid substitutions (and thus do not meet the substantial similarity definition). Pairs of antibodies, or TCRs, that recognize the same epitope can be functionally similar to each other despite whatever structural differences exist. In testing for functional similarity of immunogenicity one would generally immunize with the “altered” antigen and test the ability of the elicited response (Ab, CTL, cytokine production, etc.) to recognize the target antigen. Accordingly, two sequences may be designed to differ in certain respects while retaining the same function. Such designed sequence variants are among the embodiments of the present invention.

[0059] EXPRESSION CASSETTE—a polynucleotide sequence encoding a polypeptide, operably linked to a promoter and other transcription and translation control elements, including but not limited to enhancers, termination codons, internal ribosome entry sites, and polyadenylation sites. The cassette can also include sequences that facilitate moving it from one host molecule to another.

[0060] EMBEDDED EPITOPE—an epitope contained within a longer polypeptide, also can include an epitope in which either the N-terminus or the C-terminus is embedded such that the epitope is not in an interior position.

[0061] MATURE EPITOPE—a peptide with no additional sequence beyond that present when the epitope is bound in the MHC peptide-binding cleft.

[0062] EPITOPE CLUSTER—a polypeptide, or a nucleic acid sequence encoding it, that is a segment of a native protein sequence comprising two or more known or predicted epitopes with binding affinity for a shared MHC restriction element, wherein the density of epitopes within the cluster is greater than the density of all known or predicted epitopes with binding affinity for the shared MHC restriction element within the complete protein sequence, and as disclosed in U.S. patent application Ser. No. 09/561,571 entitled EPITOPE CLUSTERS.

[0063] SUBSTRATE OR LIBERATION SEQUENCE—a designed or engineered sequence comprising or encoding a housekeeping epitope (according to the first of the definitions offered above) embedded in a larger sequence that provides a context allowing the housekeeping epitope to be liberated by immunoproteasomal processing, directly or in combination with N-terminal trimming or other processes. terminal Degradation of cytosolic proteins takes place via the ubiquitin-dependent multi-catalytic multi-subunit protease system known as the proteasome. The proteasome degrades cytosolic proteins generating fragments that can then be translocated from the cytosol into the endoplasmic reticulum (ER) for loading onto class I MHC. Such protein fragments shall be referred to as class I peptides. The peptide loaded MHC are subsequently transported to the cell surface where they can be detected by CTL. terminal The multi-catalytic activity of the proteasome is the result of its multi-subunit structure. Subunits are expressed from different genes and assembled post-translationally into the proteasome complex. A key feature of the proteasome is its bimodal activity, which enables it to exert its protease, or cleavage function, with two discrete kinds of cleavage patterns. This bimodal action of the proteasome is extremely fundamental to understanding how CTL are targeted to recognize peripheral cells in the body and how this targeting requires synchronization between the immune system and the targeted cells.

[0064] The housekeeping proteasome is constitutively active in all peripheral cells and tissues of the body. The first mode of operation for the housekeeping proteasome is to degrade cellular protein, recycling it into amino acids. Proteasome function is therefore a necessary activity for cell life. As a corollary to its housekeeping protease activity, however, class I peptides generated by the housekeeping proteasome are presented on all of the peripheral cells of the body.

[0065] The proteasome's second mode of function is highly exclusive and occurs specifically in pAPCs or as a consequence of a cellular response to interferons (IFNs). In its second mode of activity the proteasome incorporates unique subunits, which replace the catalytic subunits of the constitutive housekeeping proteasome. This “modified” proteasome has been called the immunoproteasome, owing to its expression in pAPC and as a consequence of induction by IFN in body cells.

[0066] APC define the repertoire of CTL that recirculate through the body and are potentially active as killer cells. CTL are activated by interacting with class I peptide presented on the surface of a pAPC. Activated CTL are induced to proliferate and caused to recirculate through the body in search of diseased cells. This is why the CTL response in the body is defined specifically by the class I peptides produced by the pAPC. It is important to remember that pAPCs express the immunoproteasome, and that as a consequence of the bimodal activity of the proteasome, the cleavage pattern of proteins (and the resultant class I peptides produced) are different from those in peripheral body cells which express housekeeping proteasome. The differential proteasome activity in pAPC and peripheral body cells, therefore, is important to consider during natural infection and with therapeutic CTL vaccination strategies.

[0067] All cells of the body are capable of producing IFN in the event that they are infected by a pathogen such as a virus. IFN production in turn results in the expression of the immunoproteasome in the infected cell. Viral antigens are thereby processed by the immunoproteasome of the infected cell and the consequent peptides are displayed with class I MHC on the cell surface. At the same time, pAPC are sequestering virus antigens and are processing class I peptides with their immunoproteasome activity, which is normal for the pAPC cell type. The CTL response in the body is being stimulated specifically by the class I peptides produced by the pAPC. Fortunately, the infected cell is also producing class I peptides from the immunoproteasome, rather than the normal housekeeping proteasome. Thus, virus-related class I peptides are being produced that enable detection by the ensuing CTL response. The CTL immune response is induced by pAPC, which normally produce different class I peptides compared to peripheral body cells, owing to different proteasome activity. Therefore, during infection there is epitope synchronization between the infected cell and the immune system.

[0068] This is not the case with tumors and chronic viruses, which block the interferon system. For tumors there is no infection in the tumor cell to induce the immunoproteasome expression, and chronic virus infection either directly or indirectly blocks immunoproteasome expression. In both cases the diseased cell maintains its display of class I peptides derived from housekeeping proteasome activity and avoids effective surveillance by CTL.

[0069] In the case of therapeutic vaccination to eradicate tumors or chronic infections, the bimodal function of the proteasome and its differential activity in APC and peripheral cells of the body is significant. Upon vaccination with protein antigen, and before a CTL response can occur, the antigen must be acquired and processed into peptides that are subsequently presented on class I MHC on the pAPC surface. The activated CTL recirculate in search of cells with similar class I peptide on the surface. Cells with this peptide will be subjected to destruction by the cytolytic activity of the CTL. If the targeted diseased cell does not express the immunoproteasome, which is present in the pAPC, then the epitopes are not synchronized and CTL fail to find the desired peptide target on the surface of the diseased cell.

[0070] Preferably, therapeutic vaccine design takes into account the class I peptide that is actually present on the target tissue. That is, effective antigens used to stimulate CTL to attack diseased tissue are those that are naturally processed and presented on the surface of the diseased tissue. For tumors and chronic infection this generally means that the CTL epitopes are those that have been processed by the housekeeping proteasome. In order to generate an effective therapeutic vaccine, CTL epitopes are identified based on the knowledge that such epitopes are, in fact, produced by the housekeeping proteasome system. Once identified, these epitopes, embodied as peptides, can be used to successfully immunize or induce therapeutic CTL responses against housekeeping proteasome expressing target cells in the host.

[0071] However, in the case of DNA vaccines, there can be an additional consideration. The immunization with DNA requires that APCs take up the DNA and express the encoded proteins or peptides. It is possible to encode a discrete class I peptide on the DNA. By immunizing with this construct, APCs can be caused to express a housekeeping epitope, which is then displayed on class I MHC on the surface of the cell for stimulating an appropriate CTL response. Constructs for generation of proper termini of housekeeping epitopes have been described in U.S. patent application Ser. No. 09/561,572 entitled EXPRESSION VECTORS ENCODING EPITOPES OF TARGET-ASSOCIATED ANTIGENS, filed on Apr. 28, 2000, which is incorporated herein by reference in its entirety.

[0072] Embodiments of the invention provide expression cassettes that encode one or more embedded housekeeping epitopes, and methods for designing and testing such expression cassettes. The expression cassettes and constructs can encode epitopes, including housekeeping epitopes, derived from antigens that are associated with targets. Housekeeping epitopes can be liberated from the translation product(s) of the cassettes. For example, in some embodiments of the invention, the housekeeping epitope(s) can be flanked by arbitrary sequences or by sequences incorporating residues known to be favored in immunoproteasome cleavage sites. In further embodiments of the invention multiple epitopes can be arrayed head-to-tail. In some embodiments, these arrays can be made up entirely of housekeeping epitopes. Likewise, the arrays can include alternating housekeeping and immune epitopes. Alternatively, the arrays can include housekeeping epitopes flanked by immune epitopes, whether complete or distally truncated. In some preferred embodiments, each housekeeping epitope can be flanked on either side by an immune epitope, such that an array of such arrangements has two immune epitopes between each housekeeping epitope. Further, the arrays can be of any other similar arrangement. There is no restriction on placing a housekeeping epitope at the terminal positions of the array. The vectors can additionally contain authentic protein coding sequences or segments thereof containing epitope clusters as a source of immune epitopes.

[0073] Several disclosures make reference to polyepitopes or string-of-bead arrays. See, for example, WO0119408A1, Mar. 22, 2001; WO9955730A2, Nov. 4, 1999; WO0040261A2, Jul. 13, 2000; WO9603144A1, Feb. 8, 1996; EP1181314A1, Feb. 27, 2002; WO0123577A3, April 5; U.S. Pat No. 6,074,817, Jun. 13, 2000; U.S. Pat. No. 5,965,381, Oct. 12, 1999; WO9741440A1, Nov. 6, 1997; U.S. Pat. No. 6,130,066, October 10, 2000; U.S. Pat. No.6,004,777, December 21, 1999; U.S. Pat. No. 5,990,091, Nov. 23, 1999; WO9840501A1, Sep. 17, 1998; WO9840500A1, Sep. 17, 1998; WO018035A2, Mar. 15, 2001; WO02068654A2, Sep. 6, 2002; WO0189281A2, Nov. 29, 2001; WO0158478A, Aug. 16, 2001; EP1118860A1, Jul. 25, 2001; WO011040A1, Feb. 15, 2001; WO0073438A1, Dec. 7, 2000; WO0071158A1, Nov. 30, 2000; WO0066727A1, Nov. 9, 2000; WO0052451A1, Sep. 8, 2000; WO0052157A1, Sep. 8, 2000; WO0029008A2, May 25, 2000; WO0006723A1, Feb. 10, 2000; all of which are incorporated by reference in their entirety. Additional disclosures, all of which are hereby incorporated by reference in their entirety, include Palmowski M J, et al—J Immunol 2002;168(9):4391-8; Fang Z Y, et al—Virology 2001;291(2):272-84; Firat H, et al—J Gene Med 2002;4(1):38-45; Smith S G, et al—Clin Cancer Res 2001;7(12):4253-61; Vonderheide R H, et al—Clin Cancer Res 2001; 7(11):3343-8; Firat H, et al—Eur J Immunol 2001;31(10):3064-74; Le T T, et al—Vaccine 2001;19(32):4669-75; Fayolle C, et al—J Virol 2001;75(16):7330-8; Smith S G—Curr Opin Mol Ther 1999;1(1):10-5; Firat H, et al—Eur J Immunol 1999;29(10):3112-21; Mateo L, et al—J Immunol 1999;163(7):4058-63; Heemskerk M H, et al—Cell Immunol 1999;195(1):10-7; Woodberry T, et al—J Virol 1999;73(7):5320-5; Hanke T, et al—Vaccine 1998;16(4):426-35; Thomson S A, et al—J Immunol 1998;160(4):1717-23; Toes R E, et al—Proc Natl Acad Sci USA 1997;94(26):14660-5; Thomson S A, et al—J Immunol 1996;157(2):822-6; Thomson S A, et al—Proc Natl Acad Sci USA 1995;92(13):5845-9; Street M D, et al—Immunology 2002;106(4):526-36; Hirano K, et al—Histochem Cell Biol 2002;117(1):41-53; Ward S M, et al—Virus Genes 2001;23(1):97-104; Liu W J, et al—Virology 2000;273(2):374-82; Gariglio P, et al—Arch Med Res 1998;29(4):279-84; Suhrbier A—Immunol Cell Biol 1997;75(4):402-8; Fomsgaard A, et al—Vaccine 1999;18(7-8):681-91; An L L, et al—J Virol 1997;71(3):2292-302; Whitton J L, et al—J Virol 1993;67(1):348-52; Ripalti A, et al—J Clin Microbiol 1994;32(2):358-63; and Gilbert, S. C., et al., Nat. Biotech. 15:1280-1284, 1997.

[0074] One important feature that the disclosures in the preceding paragraph all share is their lack of appreciation for the desirability of regenerating housekeeping epitopes when the construct is expressed in a pAPC. This understanding was not apparent until the present invention. Embodiments of the invention include sequences, that when processed by an immune proteasome, liberate or generate a housekeeping epitope. Embodiments of the invention also can liberate or generate such epitopes in immunogenically effective amounts. Accordingly, while the preceding references contain disclosures relating to polyepitope arrays, none is enabling of the technology necessary to provide or select a polyepitope capable of liberating a housekeeping epitope by action of an immunoproteasome in a pAPC. In contrast, embodiments of the instant invention are based upon a recognition of the desirability of achieving this result. Accordingly, embodiments of the instant invention include any nucleic acid construct that encodes a polypeptide containing at least one housekeeping epitope provided in a context that promotes its generation via immunoproteasomal activity, whether the housekeeping epitope is embedded in a string-of-beads array or some other arrangement. Some embodiments of the invention include uses of one or more of the nucleic acid constructs or their products that are specifically disclosed in any one or more of the above-listed references. Such uses include, for example, screening a polyepitope for proper liberation context of a housekeeping epitope and/or an immune epitope, designing an effective immunogen capable of causing presentation of a housekeeping epitope and/or an immune epitope on a pAPC, immunizing a patient, and the like. Alternative embodiments include use of only a subset of such nucleic acid constructs or a single such construct, while specifically excluding one or more other such constructs, for any of the purposes disclosed herein. Some preferred embodiments employ these and/or other nucleic acid sequences encoding polyepitope arrays alone or in combination. For example, some embodiments exclude use of polyepitope arrays from one or more of the above-mentioned references. Other embodiments may exclude any combination or all of the polyepitope arrays from the above-mentioned references collectively. Some embodiments include viral and/or bacterial vectors encoding polyepitope arrays, while other embodiments specifically exclude such vectors. Such vectors can encode carrier proteins that may have some immunostimulatory effect. Some embodiments include such vectors with such immunostimulatory/immunopotentiating effects, as opposed to immunogenic effects, while in other embodiments such vectors may be included. Further, in some instances viral and bacterial vectors encode the desired epitope as a part of substantially complete proteins which are not associated with the target cell. Such vectors and products are included in some embodiments, while excluded from others. Some embodiments relate to repeated administration of vectors. In some of those embodiments, nonviral and nonbacterial vectors are included. Likewise, some embodiments include arrays that contain extra amino acids between epitopes, for example anywhere from 1-6 amino acids, or more, in some embodiments, while other embodiments specifically exclude such arrays.

[0075] Embodiments of the present invention also include methods, uses, therapies, and compositions directed to various types of targets. Such targets can include, for example, neoplastic cells such as those listed below, for example; and cells infected with any virus, bacterium, protozoan, fungus, or other agents, examples of which are listed below, in Tables 1-5, or which are disclosed in any of the references listed above. Alternative embodiments include the use of only a subset of such neoplastic cells and infected cells listed below, in Tables 1-5, or in any of the references disclosed herein, or a single one of the neoplastic cells or infected cells, while specifically excluding one or more other such neoplastic cells or infected cells, for any of the purposes disclosed herein. The following are examples of neoplastic cells that can be targeted: human sarcomas and carcinomas, e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma; leukemias, e.g., acute lymphocytic leukemia and acute myelocytic leukemia (myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroleukemia); chronic leukemia (chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia); and polycythemia vera, lymphoma (Hodgkin's disease and non Hodgkin's disease), multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, hepatocellular cancer, brain cancer, stomach cancer, liver cancer, and the like. Examples of infectious agents that infect the target cells can include the following: adenovirus, cytomegalovirus, Epstein-Barr virus, herpes simplex virus 1, herpes simplex virus 2, human herpesvirus 6, varicella-zoster virus, hepatitis B virus, hepatitis D virus, papilloma virus, parvovirus B19, polyomavirus BK, polyomavirus JC, hepatitis C virus, measles virus, rubella virus, human immunodeficiency virus (HIV), human T cell leukemia virus I, human T cell leukemia virus II, Chlamydia, Listeria, Salmonella, Legionella, Brucella, Coxiella, Rickettsia, Mycobacterium, Leishmania, Trypanasoma, Toxoplasma, Plasmodium, and the like. Exemplary infectious agents and neoplastic cells are also included in Tables 1-5 below.

[0076] Furthermore the targets can include neoplastic cells described in or cells infected by agents that are described in any of the following references: Jäger, E. et al., “Granulocyte-macrophage-colony-stimulating factor enhances immune responses to melanoma-associated peptides in vivo,” Int. J Cancer, 67:54-62 (1996); Kündig, T. M., Althage, A., Hengartner, H. & Zinkemagel, R. M., “A skin test to assess CD8+ cytotoxic T cell activity,” Proc. Natl. Acad Sci. USA, 89:7757-76 (1992); Bachmann, M. F. & Kundig, T. M., “In vitro vs. in vivo assays for the assessment of T- and B-cell function,” Curr. Opin. Immunol., 6:320-326 (1994); Kundig et al., “On the role of antigen in maintaining cytotoxic T cell memory,” Proceedings of the National Academy of Sciences of the United States of America, 93:9716-23 (1996); Steinmann, R. 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(1990); Remington, The Science and Practice of Pharmacy, Nineteenth Edition, Chapters 86-88 (1985); Cleland, Jeffery L. and Langer, Robert (Editor), “Formulation and delivery of proteins and peptides,” American Chemical Society (ACS Symposium Series, No. 567) (1994); Dickinson, Becton, which is fixed using Tegadenn transparent dressing Tegaderm™ 1624, 3M, St. Paul, Minn. 55144, USA; Santus, Giancarlo and Baker, Richard, “Osmotic drug delivery: A review of the patent literature,” Journal of Controlled Release, 35:1-21 (1995); Rammensee, U.S. Pat. No. 5,747,269, issued May 05, 1998; Magruder, U.S. Pat. No. 5,059,423, issued Oct. 22, 1991; Sandbrook, U.S. Pat. No. 4,552,651, issued Nov. 25, 1985; Eckenhoff et al., U.S. Pat. No. 3,987,790, issued Oct. 26, 1976; Theeuwes, U.S. Pat. No. 4,455,145, issued Jun. 19, 1984; Roth et al. U.S. Pat. No. 4,929,233, issued May 29 1990; van der Bruggen et al., U.S. Pat. No. 5,554,506, issued Sep. 10, 1996; Pfreundschuh, U.S. Pat. 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[0077] Additional embodiments of the invention include methods, uses, therapies, and compositions relating to a particular antigen, whether the antigen is derived from, for example, a target cell or an infective agent, such as those mentioned above. Some preferred embodiments employ the antigens listed herein, in Tables 1-5, or in the list below, alone, as subsets, or in any combination. For example, some embodiments exclude use of one or more of those antigens. Other embodiments may exclude any combination or all of those antigens. Several examples of such antigens include MelanA (MART-I), gp100 (Pmel 17), tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, CEA, RAGE, NY-ESO, SCP-1, Hom/Mel-40, PRAME, p53, H-Ras, HER-2/neu, BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein Barr virus antigens, EBNA, human papillomavirus (HPV) antigens E6 and E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, p185erbB2, p180erbB-3, c-met, nmn-23H1, PSA, TAG-72-4, CAM 17.1, NuMa, K-ras, β-Catenin, CDK4, Mum-1, p16, as well as any of those set forth in the above mentioned references. Other antigens are included in Tables 1-4 below.

[0078] Further embodiments include methods, uses, compositions, and therapies relating to epitopes, including, for example those epitopes listed in Tables 1-5. These epitopes can be useful to flank housekeeping epitopes in screening vectors, for example. Some embodiments include one or more epitopes from Tables 1-5, while other embodiments specifically exclude one or more of such epitopes or combinations thereof.

TABLE 1
		AA	T cell epitope MHC
Virus	Protein	Position	ligand (Antigen)	MHC molecule
Adenovirus 3	E3 9Kd	30-38	LIVIGILIL	HLA-A*0201
			(SEQ. ID NO.: 44)
Adenovirus 5	EIA	234-243	SGPSNTPPEI	H2-Db
			(SEQ. ID NO.: 45)
Adenovirus 5	E1B	192-200	VNIRNCCY1	H2-Db
			(SEQ. ID NO.: 46)
Adenovirus 5	EIA	234-243	SGPSNIPPEI (T > I)	H2-Db
			(SEQ. ID NO.: 47)
CSFV	NS	2276-2284	ENALLVALF	SLA,
	polyprotein			haplotype d/d
			(SEQ. ID NO.: 48)
Dengue virus 4	NS3	500-508	TPEGIIPTL	HLA-B*3501
			(SEQ. ID NO.: 49)
EBV	LMP-2	426-434	CLGGLLTMV	HLA-A*0201
			(SEQ. ID NO.: 50)
EBV	EBNA-1	480-484	NIAEGLRAL	HLA-A*0201
			(SEQ. ID NO.: 51)
EBV	EBNA-1	519-527	NLRRGTALA	HLA-A*0201
			(SEQ. ID NO.: 52)
EBV	EBNA-1	525-533	ALAIPQCRL	HLA-A*0201
			(SEQ. ID NO.: 53)
EBV	EBNA-1	575-582	VLKDAIKDL	HLA-A*0201
			(SEQ. ID NO.: 54)
EBV	EBNA-1	562-570	FMVFLQTHI	HLA-A*0201
			(SEQ. ID NO.: 55)
EBV	EBNA-2	15-23	HLIVDTDSL	HLA-A*0201
			(SEQ. ID NO.: 56)
EBV	EBNA-2	22-30	SLGNPSLSV	HLA-A*0201
			(SEQ. ID NO.: 57)
EBV	EBNA-2	126-134	PLASAMRML	HLA-A*0201
			(SEQ. ID NO.: 58)
EBV	EBNA-2	132-140	RMLWMANY1	HLA-A*0201
			(SEQ. ID NO.: 59)
EBV	EBNA-2	133-141	MLWMANYIV	HLA-A*0201
			(SEQ. ID NO.: 60)
EBV	EBNA-2	151-159	ILPQGPQTA	HLA-A*0201
			(SEQ. ID NO.: 61)
EBV	EBNA-2	171-179	PLRPTAPTI	HLA-A*0201
			(SEQ. ID NO.: 62)
EBV	EBNA-2	205-213	PLPPATLTV	HLA-A*0201
			(SEQ. ID NO.: 63)
EBV	EBNA-2	246-254	RMHLPVLHV	HLA-A*0201
			(SEQ. ID NO.: 64)
EBV	EBNA-2	287-295	PMPLPPSQL	HLA-A*0201
			(SEQ. ID NO.: 65)
EBV	EBNA-2	294-302	QLPPPAAPA	HLA-A*0201
			(SEQ. ID NO.: 66)
EBV	EBNA-2	381-389	SMPELSPVL	HLA-A*0201
			(SEQ. ID NO.: 67)
EBV	EBNA-2	453-461	DLDESWDYI	HLA-A*0201
			(SEQ. ID NO.: 68)
EBV	BZLF1	43-51	PLPCVLWPV	HLA-A*0201
			(SEQ. ID NO.: 69)
EBV	BZLF1	167-175	SLEECDSEL	HLA-A*0201
			(SEQ. ID NO.: 70)
EBV	BZLF1	176-184	EIKRYKNRV	HLA-A*0201
			(SEQ. ID NO.: 71)
EBV	BZLF1	195-203	QLLQHYREV	HLA-A*0201
			(SEQ. ID NO.: 72)
EBV	BZLF1	196-204	LLQHYREVA	HLA-A*0201
			(SEQ. ID NO.: 73)
EBV	BZLFI	217-225	LLKQMCPSL	HLA-A*0201
			(SEQ. ID NO.: 74)
EBV	BZLF1	229-237	SIIPRTPDV	HLA-A*0201
			(SEQ. ID NO.: 75)
EBV	EBNA-6	284-293	LLDFVRFMGV	HLA-A*0201
			(SEQ. ID NO.: 76)
EBV	EBNA-3	464-472	SVRDRLARL	HLA-A*0203
			(SEQ. ID NO.: 77)
EBV	EBNA-4	416-424	IVTDFSVIK	HLA-A*1101
			(SEQ. ID NO.: 78)
EBV	EBNA-4	399-408	AVFDRKSDAK	HLA-A*0201
			(SEQ. ID NO.: 79)
EBV	EBNA-3	246-253	RYSIFFDY	HLA-A24
			(SEQ. ID NO.: 80)
EBV	EBNA-6	881-889	QPRAPIRPI	HLA-B7
			(SEQ. ID NO.: 81)
EBV	EBNA-3	379-387	RPPIFIRRI	HLA-B7
			(SEQ. ID NO.: 82)
EBV	EBNA-1	426-434	EPDVPPGAI	HLA-B7
			(SEQ. ID NO.: 83)
EBV	EBNA-1	228-236	IPQCRLTPL	HLA-B7
			(SEQ. ID NO.: 84)
EBV	EBNA-1	546-554	GPGPQPGPL	HLA-B7
			(SEQ. ID NO.: 85)
EBV	EBNA-1	550-558	QPGPLRESI	HLA-B7
			(SEQ. ID NO.: 86)
EBV	EBNA-1	72-80	R.PQKRPSCI	HLA-B7
			(SEQ. ID NO.: 87)
EBV	EBNA-2	224-232	PPTPLLTVL	HLA-B7
			(SEQ. ID NO.: 88)
EBV	EBNA-2	241-249	TPSPPRMHL	HLA-B7
			(SEQ. ID NO.: 89)
EBV	EBNA-2	244-252	PPRMHLPVL	HLA-B7
			(SEQ. ID NO.: 90)
EBV	EBNA-2	254-262	VPDQSMHPL	HLA-B7
			(SEQ. ID NO.: 91)
EBV	EBNA-2	446-454	PPSIDPADL	HLA-B7
			(SEQ. ID NO.: 92)
EBV	BZLFI	44-52	LPCVLWPVL	HLA-B7
			(SEQ. ID NO.: 93)
EBV	BZLF1	222-231	CPSLDVDSII	HLA-B7
			(SEQ. ID NO.: 94)
EBV	BZLFI	234-242	TPDVLHEDL	HLA-B7
			(SEQ. ID NO.: 95)
EBV	EBNA-3	339-347	FLRGRAYGL	HLA-B8
			(SEQ. ID NO.: 96)
EBV	EBNA-3	26-34	QAKWRLQTL	HLA-B8
			(SEQ. ID NO.: 97)
EBV	EBNA-3	325-333	AYPLHEQHG	HLA-B8
			(SEQ. ID NO.: 98)
EBV	EBNA-3	158-166	YIKSFVSDA	HLA-B8
			(SEQ. ID NO.: 99)
EBV	LMP-2	236-244	RRRWRRLTV	HLA-B*2704
			(SEQ. ID NO.: 100)
EBV	EBNA-6	258-266	RRIYDLIEL	HLA-B*2705
			(SEQ. ID NO.: 101)
EBV	EBNA-3	458-466	YPLHEQHGM	HLA-B*3501
			(SEQ. ID NO.: 102)
EBV	EBNA-3	458-466	YPLHEQHGM	HLA-B*3503
			(SEQ. ID NO.: 103)
HCV	NS3	389-397	HSKKKCDEL	HLA-B8
			(SEQ. ID NO.: 104)
HCV	env E	44-51	ASRCWVAM	HLA-B*3501
			(SEQ. ID NO.: 105)
HCV	core	27-35	GQIVGGVYL	HLA-B*40012
	protein
			(SEQ. ID NO.: 106)
HCV	NSI	77-85	PPLTDFDQGW	HLA-B*5301
			(SEQ. ID NO.: 107)
HCV	core	18-27	LMGYIPLVGA	H2-Dd
	protein
			(SEQ. ID NO.: 108)
HCV	core	16-25	ADLMGYIPLV	H2-Dd
	protein
			(SEQ. ID NO.: 109)
HCV	NS5	409-424	MSYSWTGALVTPCAEE	H2-Dd
			(SEQ. ID NO.: 110)
HCV	NS1	205-213	KHPDATYSR	Papa-A06
			(SEQ. ID NO.: 111)
HCV-1	NS3	400-409	KLVALGINAV	HLA-A*0201
			(SEQ. ID NO.: 112)
HCV-1	NS3	440-448	GDFDSVIDC	Patr-B16
			(SEQ. ID NO.: 113)
HCV-1	env E	118-126	GNASRCWVA	Patr-BI6
			(SEQ. ID NO.: 114)
HCV-1	NSI	159-167	TRPPLGNWF	Patr-B13
			(SEQ. ID NO.: 115)
HCV-1	NS3	351-359	VPHPNIEEV	Patr-B13
			(SEQ. ID NO.: 116)
HCV-1	NS3	438-446	YTGDFDSVI	Patr-B01
			(SEQ. ID NO.: 117)
HCV-1	NS4	328-335	SWAIKWEY	Patr-A1 1
			(SEQ. ID NO.: 118)
HCV-1	NSI	205-213	KHPDATYSR	Patr-A04
			(SEQ. ID NO.: 119)
HCV-1	NS3	440-448	GDFDSVIDC	Patr-A04
			(SEQ. ID NO.: 120)
HIV	gp41	583-591	RYLKDQQLL	HLA_A24
			(SEQ. ID NO.: 121)
HIV	gagp24	267-275	IVGLNKIVR	HLA-A*3302
			(SEQ. ID NO.: 122)
HIV	gagp24	262-270	EIYKRWIIL	HLA-B8
			(SEQ. ID NO.: 123)
HIV	gagp24	261-269	GE1YKRWI1	HLA-B8
			(SEQ. ID NO.: 124)
HIV	gagp17	93-101	EIKDTKEAL	HLA-B8
			(SEQ. ID NO.: 125)
HIV	gp41	586-593	YLKDQQLL	HLA-B8
			(SEQ. ID NO.: 126)
HIV	gagp24	267-277	ILGLNKIVRMY	HLA-B* 1501
			(SEQ. ID NO.: 127)
HIV	gp41	584-592	ERYLKDQQL	HLA-B14
			(SEQ. ID NO.: 128)
HIV	nef	115-125	YHTQGYFPQWQ	HLA-B17
			(SEQ. ID NO.: 129)
HIV	nef	117-128	TQGYFPQWQNYT	HLA-B17
			(SEQ. ID NO.: 130)
HIV	gp120	314-322	GRAFVT1GK	HLA-B*2705
			(SEQ. ID NO.: 131)
HIV	gagp24	263-271	KRWIILGLN	HLA-B*2702
			(SEQ. ID NO.: 132)
HIV	nef	72-82	QVPLRPMTYK	HLA-B*3501
			(SEQ. ID NO.: 133)
HIV	nef	117-125	TQGYFPQWQ	HLA-B*3701
			(SEQ. ID NO.: 134)
HIV	gagp24	143-151	HQAISPRTI,	HLA-Cw*0301
			(SEQ. ID NO.: 135)
HIV	gagp24	140-151	QMVHQAISPRTL	HLA-Cw*0301
			(SEQ. ID NO.: 136)
HIV	gp120	431-440	MYAPPIGGQI	H2-Kd
			(SEQ. ID NO.: 137)
HIV	gp160	318-327	RGPGRAFVTI	H2-Dd
			(SEQ. ID NO.: 138)
HIV	gp120	17-29	MPGRAFVTI	H2-Ld
			(SEQ. ID NO.: 139)
HIV-1	RT	476-484	ILKEPVHGV	HLA-A*0201
			(SEQ. ID NO.: 140)
HIV-1	nef	190-198	AFHHVAREL	HLA-A*0201
			(SEQ. ID NO.: 141)
HIV-1	gpI60	120-128	KLTPLCVTL	HLA-A*0201
			(SEQ. ID NO.: 142)
HIV-1	gp]60	814-823	SLLNATDIAV	HLA-A*0201
			(SEQ. ID NO.: 143)
HIV-1	RT	179-187	VIYQYMDDL	HLA-A*0201
			(SEQ. ID NO.: 144)
HIV-1	gagp 17	77-85	SLYNTVATL	HLA-A*0201
			(SEQ. ID NO.: 145)
HIV-1	gp160	315-329	RGPGRAFVT1	HLA-A*0201
			(SEQ. ID NO.: 146)
HIV-1	gp41	768-778	RLRDLLLIVTR	HLA-A3
			(SEQ. ID NO.: 147)
HIV-1	nef	73-82	QVPLRPMTYK	HLA-A3
			(SEQ. ID NO.: 148)
HIV-1	gp120	36-45	TVYYGVPVWK	HLA-A3
			(SEQ. ID NO.: 149)
HIV-1	gagp17	20-29	RLRPGGKKK	HLA-A3
			(SEQ. ID NO.: 150)
HIV-1	gp120	38-46	VYYGVPVWK	HLA-A3
			(SEQ. ID NO.: 151)
HIV-1	nef	74-82	VPLRPMTYK	HLA-a*1101
			(SEQ. ID NO.: 152)
HIV-1	gagp24	325-333	AIFQSSMTK	HLA-A*1101
			(SEQ. ID NO.: 153)
HIV-1	nef	73-82	QVPLRPMTYK	HLA-A*1101
			(SEQ. ID NO.: 154)
HIV-1	nef	83-94	AAVDLSHFLKEK	HLA-A*1101
			(SEQ. ID NO.: 155)
HIV-1	gagp24	349-359	ACQGVGGPGGHK	HLA-A*1101
			(SEQ. ID NO.: 156)
HIV-1	gagp24	203-212	ETINEEAAEW	HLA-A25
			(SEQ. ID NO.: 157)
HIV-1	nef	128-137	TPGPGVRYPL	HLA-B7
			(SEQ. ID NO.: 158)
HIV-1	gagp 17	24-31	GGKKKYKL	HLA-B8
			(SEQ. ID NO.: 159)
HIV-1	gp120	2-10	RVKEKYQHL	HLA-B8
			(SEQ. ID NO.: 160)
HIV-1	gagp24	298-306	DRFYKTLRA	HLA-B 14
			(SEQ. ID NO.: 161)
HIV-1	NEF	132-147	GVRYPLTFGWCYKLV	HLA-B18
			P
			(SEQ. ID NO.: 162)
HIV-1	gagp24	265-24	KRWIILGLNK	HLA-B*2705
			(SEQ. ID NO.: 163)
HIV-1	nef	190-198	AFHHVAREL	HLA-B*5201
			(SEQ. ID NO.: 164)
EBV	EBNA-6	335-343	KEHVIQNAF	HLA-B44
			(SEQ. ID NO.: 165)
EBV	EBNA-6	130-139	EENLLDFVRF	HLA-B*4403
			(SEQ. ID NO.: 166)
EBV	EBNA-2	42-51	DTPLIPLTIF	HLA-B51
			(SEQ. ID NO.: 167)
EBV	EBNA-6	213-222	QNGALAINTF	HLA-1362
			(SEQ. ID NO.: 168)
EBV	EBNA-3	603-611	RLRAEAGVK	HLA-A3
			(SEQ. ID NO.: 169)
HBV	sAg	348-357	GLSPTVWLSV	HLA-A*0201
			(SEQ. ID NO.: 170)
HBV	SAg	335-343	WLSLLVPFV	HLA-A*0201
			(SEQ. ID NO.: 171)
HBV	cAg	18-27	FLPSDFFPSV	HLA-A*0201
			(SEQ. ID NO.: 172)
HBV	cAg	18-27	FLPSDFFPSV	HLA-A*0202
			(SEQ. ID NO.: 173)
HBV	cAg	18-27	FLPSDFFPSV	HLA-A*0205
			(SEQ. ID NO.: 174)
HBV	cAg	18-27	FLPSDFFPSV	HLA-A*0206
			(SEQ. ID NO.: 175)
HBV	pol	575-583	FLLSLGIHlL	HLA-A*0201
			(SEQ. ID NO.: 176)
HBV	pol	816-824	SLYADSPSV	HLA-A*0201
			(SEQ. ID NO.: 177)
HBV	pol	455-463	GLSRYVARL	HLA-A*0201
			(SEQ. ID NO.: 178)
HBV	env	338-347	LLVPFVQWFV	HLA-A*0201
			(SEQ. ID NO.: 179)
HBV	pol	642-650	ALMPLYACI	HLA-A*0201
			(SEQ. ID NO.: 180)
HBV	env	378-387	LLPIFFCLWV	HLA-A*0201
			(SEQ. ID NO.: 181)
HBV	pol	538-546	YMDDVVLGA	HLA-A*0201
			(SEQ. ID NO.: 182)
HBV	env	250-258	LLLCLIFLL	HLA-A*0201
			(SEQ. ID NO.: 183)
HBV	env	260-269	LLDYQGMLPV	HLA-A*0201
			(SEQ. ID NO.: 184)
HBV	env	370-379	SIVSPFIPLL	HLA-A*0201
			(SEQ. ID NO.: 185)
HBV	env	183-191	FLLTRILTI	HLA-A*0201
			(SEQ. ID NO.: 186)
HBV	cAg	88-96	YVNVNMGLK	HLA-A* 1101
			(SEQ. ID NO.: 187)
HBV	cAg	141-151	STLPETTVVRR	HLA-A*3101
			(SEQ. ID NO.: 188)
HBV	cAg	141-151	STLPETTVVRR	HLA-A*6801
			(SEQ. ID NO.: 189)
HBV	cAg	18-27	FLPSDFFPSV	HLA-A*6801
			(SEQ. ID NO.: 190)
HBV	sAg	28-39	IPQSLDSWWTSL	H2-Ld
			(SEQ. ID NO.: 191)
HBV	cAg	93-100	MGLKFRQL	H2-Kb
			(SEQ. ID NO.: 192)
HBV	preS	141-149	STBXQSGXQ	HLA-A*0201
			(SEQ. ID NO.: 193)
HCMV	gp B	618-628	FIAGNSAYEYV	HLA-A*0201
			(SEQ. ID NO.: 194)
HCMV	E1	978-989	SDEEFAIVAYTL	HLA-B18
			(SEQ. ID NO.: 195)
HCMV	pp65	397-411	DDVWTSGSDSDEELV	HLA-b35
			(SEQ. ID NO.: 196)
HCMV	pp65	123-131	IPSINVHHY	HLA-B*3501
			(SEQ. ID NO.: 197)
HCMV	pp65	495-504	NLVPMVATVO	HLA-A*0201
			(SEQ. ID NO.: 198)
HCMV	pp65	415-429	RKTPRVTOGGAMAGA	HLA-B7
			(SEQ. ID NO.: 199)
HCV	MP	17-25	DLMGYIPLV	HLA-A*0201
			(SEQ. ID NO.: 200)
HCV	MP	63-72	LLALLSCLTV	HLA-A*0201
			(SEQ. ID NO.: 201)
HCV	MP	105-112	ILHTPGCV	HLA-A*0201
			(SEQ. ID NO.: 202)
HCV	env E	66-75	QLRRHIDLLV	HLA-A*0201
			(SEQ. ID NO.: 203)
HCV	env E	88-96	DLCGSVFLV	HLA-A*0201
			(SEQ. ID NO.: 204)
HCV	env E	172-180	SMVGNWAKV	HLA-A*0201
			(SEQ. ID NO.: 205)
HCV	NSI	308-316	HLIIQNIVDV	HLA-A*0201
			(SEQ. ID NO.: 206)
HCV	NSI	340-348	FLLLADARV	HLA-A*0201
			(SEQ. ID NO.: 207)
HCV	NS2	234-246	GLRDLAVAVEPVV	HLA-A*0201
			(SEQ. ID NO.: 208)
HCV	NSI	18-28	SLLAPGAKQNV	HLA-A*0201
			(SEQ. ID NO.: 209)
HCV	NSI	19-28	LLAPGAKQNV	HLA-A*0201
			(SEQ. ID NO.: 210)
HCV	NS4	192-201	LLFNILGGWV	HLA-A*0201
			(SEQ. ID NO.: 211)
HCV	NS3	579-587	YLVAYQATV	HLA-A*0201
			(SEQ. ID NO.: 212)
HCV	core	34-43	YLLPRRGPRL	HLA-A*0201
	protein
			(SEQ. ID NO.: 213)
HCV	MP	63-72	LLALLSCLTI	HLA-A*0201
			(SEQ. ID NO.: 214)
HCV	NS4	174-182	SLMAFTAAV	HLA-A*0201
			(SEQ. ID NO.: 215)
HCV	NS3	67-75	CINGVCWTV	HLA-A*0201
			(SEQ. ID NO.: 216)
HCV	NS3	163-171	LLCPAGHAV	HLA-A*0201
			(SEQ. ID NO.: 217)
HCV	NS5	239-247	ILDSFDPLV	HLA-A*0201
			(SEQ. ID NO.: 218)
HCV	NS4A	236-244	ILAGYGAGV	HLA-A*0201
			(SEQ. ID NO.: 219)
HCV	NS5	714-722	GLQDCTMLV	HLA-A*0201
			(SEQ. ID NO.: 220)
HCV	NS3	281-290	TGAPVTYSTY	HLA-A*0201
			(SEQ. ID NO.: 221)
HCV	NS4A	149-157	HMWNFISGI	HLA-A*0201
			(SEQ. ID NO.: 222)
HCV	NS5	575-583	RVCEKMALY	HLA-A*0201-A3
			(SEQ. ID NO.: 223)
HCV	NS1	238-246	TINYTIFK	HLA-A*1101
			(SEQ. ID NO.: 224)
HCV	NS2	109-116	YISWCLWW	HLA-A23
			(SEQ. ID NO.: 225)
HCV	core	40-48	GPRLGVRAT	HLA-B7
	protein
			(SEQ. ID NO.: 226)
HIV-1	gp120	380-388	SFNCGGEFF	HLA-Cw*0401
			(SEQ. ID NO.: 227)
HIV-1	RT	206-214	TEMEKEGKI	H2-Kk
			(SEQ. ID NO.: 228)
HIV-1	p17	18-26	KIRLRPGGK	HLA-A*0301
			(SEQ. ID NO.: 229)
HIV-1	P17	20-29	RLRPGGKKKY	HLA-A*0301
			(SEQ. ID NO.: 230)
HIV-1	RT	325-333	AIFQSSMTK	HLA-A*0301
			(SEQ. ID NO.: 231)
HIV-1	p17	84-92	TLYCVHQRI	HLA-A11
			(SEQ. ID NO.: 232)
HIV-1	RT	508-517	IYQEPFKNLK	HLA-A11
			(SEQ. ID NO.: 233)
HIV-1	p17	28-36	KYKLKHIVW	HLA-A24
			(SEQ. ID NO.: 234)
HIV-1	gp120	53-62	LFCASDAKAY	HLA-A24
			(SEQ. ID NO.: 235)
HIV-1	gagp24	145-155	QAISPRTLNAW	HLA-A25
			(SEQ. ID NO.: 236)
HIV-1	gagp24	167-175	EVIPMFSAL	HLA-A26
			(SEQ. ID NO.: 237)
HIV-1	RT	593-603	ETFYVDGAANR	HLA-A26
			(SEQ. ID NO.: 238)
HIV-1	gp41	775-785	RLRDLLLIVTR	HLA-A31
			(SEQ. ID NO.: 239)
HIV-1	RT	559-568	PIQKETWETW	HLA-A32
			(SEQ. ID NO.: 240)
HIV-1	gp120	419-427	RIKQIINMW	HLA-A32
			(SEQ. ID NO.: 241)
HIV-1	RT	71-79	ITLWQRPLV	HLA-A*6802
			(SEQ. ID NO.: 242)
HIV-1	RT	85-93	DTVLEEMNL	HLA-A*6802
			(SEQ. ID NO.: 243)
HIV-1	RT	71-79	ITLWQRPLV	HLA-A*7401
			(SEQ. ID NO.: 244)
HIV-1	gag p24	148-156	SPRTLNAWV	HLA-B7
			(SEQ. ID NO.: 245)
HIV-1	gagp24	179-187	ATPQDLNTM	HLA-B7
			(SEQ. ID NO.: 246)
HIV-1	gp120	303-312	RPNNNTRKSI	HLA-B7
			(SEQ. ID NO.: 247)
HIV-1	gp41	843-851	IPRRIRQGL	HLA-B7
			(SEQ. ID NO.: 248)
HIV-1	p17	74-82	ELRSLYNTV	HLA-B8
			(SEQ. ID NO.: 249)
HIV-1	nef	13-20	WPTVRERM	HLA-B8
			(SEQ. ID NO.: 250)
HIV-1	nef	90-97	FLKEKGGL	HLA-B8
			(SEQ. ID NO.: 251)
HIV-1	gag p24	183-191	DLNTMLNTV	HLA-B14
			(SEQ. ID NO.: 252)
HIV-1	P17	18-27	KIRLRPGGKK	HLA-B27
			(SEQ. ID NO.: 253)
HIV-1	p17	19-27	IRLRPGGKK	HLA-B27
			(SEQ. ID NO.: 254)
HIV-1	gp41	791-799	GRRGWEALKY	HLA-B27
			(SEQ. ID NO.: 255)
HIV-1	nef	73-82	QVPLRPMTYK	HLA-B27
			(SEQ. ID NO.: 256)
HIV-1	GP41	590-597	RYLKDQQL	HLA-B27
			(SEQ. ID NO.: 257)
HIV-1	nef	105-114	RRQDILDLWI	HLA-B*2705
			(SEQ. ID NO.: 258)
HIV-1	nef	134-141	RYPLTFGW	HLA-B*2705
			(SEQ. ID NO.: 259)
HIV-1	p17	36-44	WASRELERF	HLA-B35
			(SEQ. ID NO.: 260)
HIV-1	GAG P24	262-270	TVLDVGDAY	HLA-B35
			(SEQ. ID NO.: 261)
HIV-1	gp120	42-52	VPVWKEATTTL	HLA-B35
			(SEQ. ID NO.: 262)
HIV-1	P17	36-44	NSSKVSQNY	HLA-B35
			(SEQ. ID NO.: 263)
HIV-1	gag p24	254-262	PPIPVGDIY	HLA-B35
			(SEQ. ID NO.: 264)
HIV-1	RT	342-350	HPDIVIYQY	HLA-B35
			(SEQ. ID)NO.: 265)
HIV-1	gp41	611-619	TAVPWNASW	HLA-B35
			(SEQ. ID NO.: 266)
HIV-1	gag	245-253	NPVPVGN1Y	HLA-B35
			(SEQ. ID NO.: 267)
HIV-1	nef	120-128	YFPDWQNYT	HLA-B37
			(SEQ. ID NO.: 268)
HIV-1	gag p24	193-201	GHQAAMQML	HLA-B42
			(SEQ. ID NO.: 269)
HIV-1	p17	20-29	RLRPGGKKKY	HLA-B42
			(SEQ. ID NO.: 270)
HIV-1	RT	438-446	YPGIKVRQL	HLA-B42
			(SEQ. ID NO.: 271)
HIV-1	RT	591-600	GAETFYVDGA	HLA-B45
			(SEQ. ID NO.: 272)
HIV-1	gag p24	325-333	NANPDCKTI	HLA-B51
			(SEQ. ID NO.: 273)
HIV-1	gag p24	275-282	RMYSPTSI	HLA-B52
			(SEQ. ID NO.: 274)
HIV-1	gp120	42-51	VPVWKEATTT	HLA-B*5501
			(SEQ. ID NO.: 275)
HIV-1	gag p24	147-155	ISPRTLNAW	HLA-B57
			(SEQ. ID NO.: 276)
HIV-1	gag p24	240-249	TSTLQEQIGW	HLA-B57
			(SEQ. ID NO.: 277)
HIV-1	gag p24	162-172	KAFSPEVIPMF	HLA-B57
			(SEQ. ID NO.: 278)
HIV-1	gag p24	311-319	QASQEVKNW	HLA-B57
			(SEQ. ID NO.: 279)
HIV-1	gag p24	311-319	QASQDVKNW	HLA-B57
			(SEQ. ID NO.: 280)
HIV-1	nef	116-125	HTQGYFPDWQ	HLA-B57
			(SEQ. ID NO.: 281)
HIV-1	nef	120-128	YFPDWQNYT	HLA-B57
			(SEQ. ID NO.: 282)
HIV-1	gag p24	240-249	TSTLQEQIGW	HLA-B58
			(SEQ. ID NO.: 283)
HIV-1	p17	20-29	RLRPGGKKKY	HLA-B62
			(SEQ. ID NO.: 284)
HIV-1	p24	268-277	LGLNKJVRMY	HLA-B62
			(SEQ. ID NO.: 285)
HIV-1	RT	415-426	LVGKLNWASQIY	HLA-B62
			(SEQ. ID NO.: 286)
HIV-1	RT	476-485	ILKEPVHGVY	HLA-B62
			(SEQ. ID NO.: 287)
HIV-1	nef	117-127	TQGYFPDWQNY	HLA-B62
			(SEQ. ID NO.: 288)
HIV-1	nef	84-91	AVDLSHFL	HLA-B62
			(SEQ. ID NO.: 289)
HIV-1	gag p24	168-175	VIPMFSAL	HLA-Cw*0102
			(SEQ. ID NO.: 290)
HIV-1	gp120	376-384	FNCGGEFFY	HLA-A29
			(SEQ. ID NO.: 291)
HIV-1	gp120	375-383	SFNCGGEFF	HLA-B15
			(SEQ. ID NO.: 292)
HIV-1	nef	136-145	PLTFGWCYKL	HLA-A*0201
			(SEQ. ID NO.: 293)
HIV-1	nef	180-189	VLEWRFDSRL	HLA-A*0201
			(SEQ. ID NO.: 294)
HIV-1	nef	68-77	FPVTPQVPLR	HLA-B7
			(SEQ. ID NO.: 295)
HIV-1	nef	128-137	TPGPGVRYPL	HLA-B7
			(SEQ. ID NO.: 296)
HIV-1	gag p24	308-316	QASQEVKNW	HLA-Cw*0401
			(SEQ. ID NO.: 297)
HIV-1 IIIB	RT	273-282	VPLDEDFRKY	HLA-B35
			(SEQ. ID NO.: 298)
HIV-1 IIIB	RT	25-33	NPDIVIYQY	HLA-B35
			(SEQ. ID NO.: 299)
HIV-1 IIIB	gp41	557-565	RAIEAQAHL	HLA-B51
			(SEQ. ID NO.: 300)
HIV-1 IIIB	RT	231-238	TAFTIPSI	HLA-B51
			(SEQ. ID NO.: 301)
HIV-1 IIIB	p24	215-223	VHPVHAGPIA	HLA-B*5501
			(SEQ. ID NO.: 302)
HIV-1 IIIB	gp120	156-165	NCSFNISTSI	HLA-Cw8
			(SEQ. ID NO.: 303)
HIV-1 IIIB	gp120	241-249	CTNVSTVQC	HLA-Cw8
			(SEQ. ID NO.: 304)
HIV-1 5F2	gp120	312-320	IGPGRAFHT	H2-Dd
			(SEQ. ID NO.: 305)
HIV-1 5F2	pol	25-33	NPDIVIYQY	HLA-B*3501
			(SEQ. ID NO.: 306)
HIV-1 5F2	pol	432-441	EPIVGAETFY	HLA-B*3501
			(SEQ. ID NO.: 307)
HIV-1 5F2	pol	432-440	EPIVGAETF	HLA-B*3501
			(SEQ. ID NO.: 308)
HIV-1 5F2	pol	6-14	SPAIFQSSM	HLA-B*3501
			(SEQ. ID NO.: 309)
HIV-1 5F2	pol	59-68	VPLDKDFRKY	HLA-B*3501
			(SEQ. ID NO.: 310)
HIV-1 5F2	pol	6-14	IPLTEEAEL	HLA-B*3501
			(SEQ. ID NO.: 311)
HIV-1 5F2	nef	69-79	RPQVPLRPMTY	HLA-B*3501
			(SEQ. ID NO.: 312)
HIV-1 5F2	nef	66-74	FPVRPQVPL	HLA-B*3501
			(SEQ. ID NO.: 313)
HIV-1 5F2	env	10-18	DPNPQEVVL	HLA-B*3501
			(SEQ. ID NO.: 314)
HIV-1 5F2	env	7-15	RPIVSTQLL	HLA-B*3501
			(SEQ. ID NO.: 315)
HIV-1 5F2	pol	6-14	IPLTEEAEL	HLA-B51
			(SEQ. ID NO.: 316)
HIV-1 5F2	env	10-18	DPNPQEVVL	HLA-B51
			(SEQ. ID NO.: 317)
HIV-1 5F2	gagp24	199-207	AMQMLKETI	H2-Kd
			(SEQ. ID NO.: 318)
HIV-2	gagp24	182-190	TPYDrNQML	HLA-B*5301
			(SEQ. ID NO.: 319)
HIV-2	gag	260-269	RRWIQLGLQKV	HLA-B*2703
			(SEQ. ID NO.: 320)
HIV-1 5F2	gp41	593-607	GIWGCSGKLICTTAV	HLA-B17
			(SEQ. ID NO.: 321)
HIV-1 5F2	gp41	753-767	ALIWEDLRSLCLFSY	HLA-B22
			(SEQ. ID NO.: 322)
HPV 6b	E7	21-30	GLHCYEQLV	HLA-A*0201
			(SEQ. ID NO.: 323)
HPV 6b	E7	47-55	PLKQHFQIV	HLA-A*0201
			(SEQ. ID NO.: 324)
HPV11	E7	4-12	RLVTLKDIV	HLA-A*0201
			(SEQ. ID NO.: 325)
HPV16	E7	86-94	TLGIVCPIC	HLA-A*0201
			(SEQ. ID NO.: 326)
HPV16	E7	85-93	GTLGIVCPI	HLA-A*0201
			(SEQ. ID NO.: 327)
HPV16	E7	12-20	MLDLQPETT	HLA-A*0201
			(SEQ. ID NO.: 328)
HPV16	E7	11-20	YMLDLQPETT	HLA-A*0201
			(SEQ. ID NO.: 329)
HPV16	E6	15-22	RPRKLPQL	HLA-B7
			(SEQ. ID NO.: 330)
HPV16	E6	49-57	RAHYNIVTF	HW-Db
			(SEQ. ID NO.: 331)
HSV	gp B	498-505	SSIEFARL	H2-Kb
			(SEQ. ID NO.: 332)
HSV-1	gp C	480-488	GIGIGVLAA	HLA-A*0201
			(SEQ. ID NO.: 333)
HSV-1	ICP27	448-456	DYATLGVGV	H2-Kd
			(SEQ. ID NO.: 334)
HSV-1	ICP27	322-332	LYRTFAGNPRA	H2-Kd
			(SEQ. ID NO.: 335)
HSV-1	UL39	822-829	QTFDFGRL	H2-Kb
			(SEQ.ID NO.: 336)
HSV-2	gpC	446-454	GAGIGVAVL	HLA-A*0201
			(SEQ. ID NO.: 337)
HLTV-1	TAX	11-19	LLFGYPVYV	HLA-A*0201
			(SEQ. ID NO.: 338)
Influenza	MP	58-66	GILGFVFTL	HLA-A*0201
			(SEQ. ID NO.: 339)
Influenza	MP	59-68	ILGFVFTLTV	HLA-A*0201
			(SEQ. ID NO.: 340)
Influenza	NP	265-273	ILRGSVAHK	HLA-A3
			(SEQ. ID NO.: 341)
Influenza	NP	91-99	KTGGPIYKR	HLA-A*6801
			(SEQ. ID NO.: 342)
Influenza	NP	380-388	ELRSRYWAI	HLA-B8
			(SEQ. ID NO.: 343)
Influenza	NP	381-388	LRSRYWAI	HLA-B*2702
			(SEQ. ID NO.: 344)
Influenza	NP	339-347	EDLRVLSFI	HLA-B*3701
			(SEQ. ID NO.: 345)
Influenza	NSI	158-166	GEISPLPSL	HLA-B44
			(SEQ. ID NO.: 346)
Influenza	NP	338-346	FEDLRVLSF	HLA-B44
			(SEQ. ID NO.: 347)
Influenza	NSI	158-166	GEISPLPSL	HLA-B*4402
			(SEQ. ID NO.: 348)
Influenza	NP	338-346	FEDLRVLSF	HLA-B*4402
			(SEQ. ID NO.: 349)
Influenza	PBI	591-599	VSDGGPKLY	HLA-A1
			(SEQ. ID NO.: 350)
Influenza A	NP	44-52	CTELKLSDY	HLA-A1
			(SEQ. ID NO.: 351)
Influenza	NSI	122-130	AIMDKNIIL	HLA-A*0201
			(SEQ. ID NO.: 352)
Influenza A	NSI	123-132	IMDKNIILKA	HLA-A*0201
			(SEQ. ID NO.: 353)
Influenza A	NP	383-391	SRYWAIRTR	HLA-B*2705
			(SEQ. ID NO.: 354)
Influenza A	NP	147-155	TYQRTRALV	H2-Kd
			(SEQ. ID NO.: 355)
Influenza A	HA	210-219	TYVSVSTSTL	H2-Kd
			(SEQ. ID NO.: 356)
Influenza A	HA	518-526	IYSTVASSL	H2-Kd
			(SEQ. ID NO.: 357)
Influenza A	HA	259-266	FEANGNLI	H2-Kk
			(SEQ. ID NO.: 358)
Influenza A	HA	10-18	IEGGWTGM1	H2-Kk
			(SEQ. ID NO.: 359)
Influenza A	NP	50-57	SDYEGRLI	H2-Kk
			(SEQ. ID NO.: 360)
Influenza a	NSI	152-160	EEGAIVGEI	H2-Kk
			(SEQ. ID NO.: 361)
Influenza A34	NP	336-374	ASNENMETM	H2Db
			(SEQ. ID NO.: 362)
Influenza A68	NP	366-374	ASNENMDAM	H2Db
			(SEQ. ID NO.: 363)
Influenza B	NP	85-94	KLGEFYNQMM	HLA-A*0201
			(SEQ. ID NO.: 364)
Influenza B	NP	85-94	KAGEFYNQMM	HLA-A*0201
			(SEQ. ID NO.: 365)
Influenza JAP	HA	204-212	LYQNVGTYV	H2Kd
			(SEQ. ID NO.: 366)
Influenza JAP	HA	210-219	TYVSVGTSTL	H2-Kd
			(SEQ. ID NO.: 367)
Influenza JAP	HA	523-531	VYQILATYA	H2-Kd
			(SEQ. ID NO.: 368)
Influenza JAP	HA	529-537	IYATVAGSL	H2-Kd
			(SEQ. ID NO.: 369)
Influenza JAP	HA	210-219	TYVSVGTSTI(L>I)	H2-Kd
			(SEQ. ID NO.: 370)
Influenza JAP	HA	255-262	FESTGNLI	H2-Kk
			(SEQ. ID NO.: 371)
JHMV	cAg	318-326	APTAGAFFF	H2-Ld
			(SEQ. ID NO.: 372)
LCMV	NP	118-126	RPQASGVYM	H2-Ld
			(SEQ. ID NO.: 373)
LCMV	NP	396-404	FQPQNGQFI	H2-Db
			(SEQ. ID NO.: 374)
LCMV	GP	276-286	SGVENPGGYCL	H2-Db
			(SEQ. ID NO.: 375)
LCMV	GP	33-42	KAVYNFATCG	H2-Db
			(SEQ. ID NO.: 376)
MCMV	pp89	168-176	YPHFMPTNL	H2-Ld
			(SEQ. ID NO.: 377)
MHV	spike	510-518	CLSWNGPHL	H2-Db
	protein
			(SEQ. ID NO.: 378)
MMTV	env gp 36	474-482	SFAVATTAL	H2-Kd
			(SEQ. ID NO.: 379)
MMTV	gag p27	425-433	SYETFISRL	H2-Kd
			(SEQ. ID NO.: 380)
MMTV	env gp73	544-551	ANYDFICV	H2-Kb
			(SEQ. ID NO.: 381)
MuLV	env p15E	574-581	KSPWFTTL	H2-Kb
			(SEQ. ID NO.: 382)
MuLV	env gp70	189-196	SSWDFITV	H2-Kb
			(SEQ. ID NO.: 383)
MuLV	gag 75K	75-83	CCLCLTVFL	H2-Db
			(SEQ. ID NO.: 384)
MuLV	env gp70	423-431	SPSYVYHQF	H2Ld
			(SEQ. ID NO.: 385)
MV	F protein	437-447	SRRYPDAVYLH	HLA-B*2705
			(SEQ. ID NO.: 386)
Mv	F protein	438-446	RRYPDAVYL	HLA-B*2705
			(SEQ. ID NO.: 387)
Mv	NP	281-289	YPALGLHEF	H2-Ld
			(SEQ. ID NO.: 388)
Mv	HA	343-351	DPVIDRLYL	H2-Ld
			(SEQ. ID NO.: 389)
MV	HA	544-552	SPGRSFSYF	H2-Ld
			(SEQ. ID NO.: 390)
Poliovirus	VP1	111-118	TYKDTVQL	H2-kd
			(SEQ. ID NO.: 391)
Poliovirus	VP1	208-217	FYDGFSKVPL	H2-Kd
			(SEQ. ID NO.: 392)
Pseudorabies	G111	455-463	IAGIGILAI	HLA-A*0201
virus gp
			(SEQ. ID NO.: 393)
Rabiesvirus	NS	197-205	VEAEIAHQI	H2-Kk
			(SEQ. ID NO.: 394)
Rotavirus	VP7	33-40	11YRFLL1	H2-Kb
			(SEQ. ID NO.: 395)
Rotavirus	VP6	376-384	VGPVFPPGM	H2-Kb
			(SEQ. ID NO.: 396)
Rotavirus	VP3	585-593	YSGYIFRDL	H2-Kb
			(SEQ. ID NO.: 397)
RSV	M2	82-90	SYIGSINNI	H2-Kd
			(SEQ. ID NO.: 398)
SIV	gagp11C	179-190	EGCTPYDTNQML	Mamu-A*01
			(SEQ. ID NO.: 399)
SV	NP	324-332	FAPGNYPAL	H2-Db
			(SEQ. ID NO.: 400)
SV	NP	324-332	FAPCTNYPAL	H2-Kb
			(SEQ. ID NO.: 401)
SV40	T	404-411	VVYDFLKC	H2-Kb
			(SEQ. ID NO.: 402)
SV40	T	206-215	SAINNYAQKL	H2-Db
			(SEQ. ID NO.: 403)
SV40	T	223-231	CKGVNKEYL	H2-Db
			(SEQ. ID NO.: 404)
SV40	T	489-497	QGINNLDNL	H2-Db
			(SEQ. ID NO.: 405)
SV40	T	492-500	NNLDNLRDY(L)	H2-Db
		(501)
			(SEQ. ID NO.: 406)
SV40	T	560-568	SEFLLEKRI	H2-Kk
			(SEQ. ID NO.: 407)
VSV	NP	52-59	RGYVYQGL	H2-Kb
			(SEQ. ID NO.: 408)

[0079]

TABLE 2
HLA-A1	Position (Antigen)	Source
T cell	EADPTGHSY	MAGE-1 161-169
epitopes
	(SEQ. ID NO.: 409)
	VSDGGPNLY	Influenza A PB 1591-599
	(SEQ. ID NO.: 410)
	CTELKLSDY	Influenza A NP 44-52
	(SEQ. ID NO.: 411)
	EVDPIGHLY	MAGE-3 168-176
	(SEQ. ID NO.: 412)
HLA-A201	MLLSVPLLLG	Calreticulin signal
		sequence I-10
	(SEQ. ID NO.: 413)
	STBXQSGXQ	HBV PRE-S PROTEIN 141-149
	(SEQ. ID NO.: 414)
	YMDGTMSQV	Tyrosinase 369-377
	(SEQ. ID NO.: 415)
	ILKEPVHGV	HIV-I RT 476-484
	(SEQ. ID NO.: 416)
	LLGFVFTLTV	Influenza MP 59-68
	(SEQ. ID NO.: 417)
	LLFGYPVYVV	HTLV-1 tax 11-19
	(SEQ. ID NO.: 418)
	GLSPTVWLSV	HBV sAg 348-357
	(SEQ. ID NO.: 419)
	WLSLLVPFV	HBV sAg 335-343
	(SEQ. ID NO.: 420)
	FLPSDFFPSV	HBV cAg 18-27
	(SEQ. ID NO.: 421)
	CLGOLLTMV	EBV LMP-2 426-434
	(SEQ. ID NO.: 422)
	FLAGNSAYEYV	HCMV gp 618-628B
	(SEQ. ID NO.: 423)
	KLGEFYNQMM	Influenza BNP 85-94
	(SEQ. ID NO.: 424)
	KLVALGINAV	HCV-1 NS3 400-409
	(SEQ. ID NO.: 425)
	DLMGYIPLV	HCV MP 17-25
	(SEQ. ID NO.: 426)
	RLVTLKDIV	HPV 11 EZ 4-12
	(SEQ. ID NO.: 427)
	MLLAVLYCL	Tyrosinase 1-9
	(SEQ. ID NO.: 428)
	AAGIGILTV	Melan A\Mart-127-35
	(SEQ. ID NO.: 429)
	YLEPGPVTA	Pmel 17/gp 100 480-488
	(SEQ. ID NO.: 430)
	ILDGTATLRL	Pmel 17/gp 100 457-466
	(SEQ. ID NO.: 431)
	LLDGTATLRL	Pmel gplOO 457-466
	(SEQ. ID NO.: 432)
	ITDQVPFSV	Pmel gp 100 209-217
	(SEQ. ID NO.: 433)
	KTWGQYWQV	Pmel gp 100 154-162
	(SEQ. ID NO.: 434)
	TITDQVPFSV	Pmel gp 100 208-217
	(SEQ. ID NO.: 435)
	AFHIIVAREL	HIV-I nef 190-198
	(SEQ. ID NO.: 436)
	YLNKIQNSL	P. falciparum CSP 334-342
	(SEQ. ID NO.: 437)
	MMRKLAELSV	P. falciparum CSP 1-10
	(SEQ. ID NO.: 438)
	KAGEFYNQMM	Influenza BNP 85-94
	(SEQ. ID NO.: 439)
	NIAEGLRAL	EBNA-1 480-488
	(SEQ. ID NO.: 440)
	NLRRGTALA	EBNA-1 519-527
	(SEQ. ID NO.: 441)
	ALAIPQCRL	EBNA-1 525-533
	(SEQ. ID NO.: 442)
	VLKDAIKDL	EBNA-1 575-582
	(SEQ. ID NO.: 443)
	FMVFLQTHI	EBNA-1 562-570
	(SEQ. ID NO.: 444)
	HLIVDTDSL	EBNA-2 15-23
	(SEQ. ID NO.: 445)
	SLGNPSLSV	EBNA-2 22-30
	(SEQ. ID NO.: 446)
	PLASAMRML	EBNA-2 126-134
	(SEQ. ID NO.: 447)
	RMLWMANYI	EBNA-2 132-140
	(SEQ. ID NO.: 448)
	MLWMANYIV	EBNA-2 133-141
	(SEQ. ID NO.: 449)
	ILPQGPQTA	EBNA-2 151-159
	(SEQ. ID NO.: 450)
	PLRPTAPTTI	EBNA-2 171-179
	(SEQ. ID NO.: 451)
	PLPPATLTV	EBNA-2 205-213
	(SEQ. ID NO.: 452)
	RMHLPVLHV	EBNA-2 246-254
	(SEQ. ID NO.: 453)
	PMPLPPSQL	EBNA-2 287-295
	(SEQ. ID NO.: 454)
	QLPPPAAPA	EBNA-2 294-302
	(SEQ. ID NO.: 455)
	SMPELSPVL	EBNA-2 381-389
	(SEQ. ID NO.: 456)
	DLDESWDY1	EBNA-2 453-461
	(SEQ. ID NO.: 457)
	PLPCVLWPVV	BZLF1 43-51
	(SEQ. ID NO.: 458)
	SLEECDSEL	BZLF1 167-175
	(SEQ. ID NO.: 459)
	EIKRYKNRV	BZLF1 176-184
	(SEQ. ID NO.: 460)
	QLLQFIYREV	BZLF1 195-203
	(SEQ. ID NO.: 461)
	LLQHYREVA	BZLFI 196-204
	(SEQ. ID NO.: 462)
	LLKQMCPSL	BZLFI 217-225
	(SEQ. ID NO.: 463)
	SIIPRTPDV	BZLFI 229-237
	(SEQ. ID NO.: 464)
	AIMDKNIIL	Influenza A NSI 122-130
	(SEQ. ID NO.: 465)
	IMDKNIILKA	Influenza A NSI 123-132
	(SEQ. ID NO.: 466)
	LLALLSCLTV	HCV MP 63-72
	(SEQ. ID NO.: 467)
	ILHTPGCV	HCV MP 105-112
	(SEQ. ID NO.: 468)
	QLRRHIDLLV	HCV env E 66-75
	(SEQ. ID NO.: 469)
	DLCGSVFLV	HCV env E 88-96
	(SEQ. ID NO.: 470)
	SMVGNWAKV	HCV env E 172-180
	(SEQ. ID NO.: 471)
	HLHQNIVDV	HCV NSI 308-316
	(SEQ. ID NO.: 472)
	FLLLADARV	HCV NSI 340-348
	(SEQ. ID NO.: 473)
	GLRDLAVAVEPVV	HCV NS2 234-246
	(SEQ. ID NO.: 474)
	SLLAPGAKQNV	HCV NS1 18-28
	(SEQ. ID NO.: 475)
	LLAPGAKQNV	HCV NS1 19-28
	(SEQ. ID NO.: 476)
	FLLSLGIHL	HBV pol 575-583
	(SEQ. ID NO.: 477)
	SLYADSPSV	HBV pol 816-824
	(SEQ. ID NO.: 478)
	GLSRYVARL	HBV POL 455-463
	(SEQ. ID NO.: 479)
	KIFGSLAFL	HER-2 369-377
	(SEQ. ID NO.: 480)
	ELVSEFSRM	HER-2 971-979
	(SEQ. ID NO.: 481)
	KLTPLCVTL	HIV-I gp 160 120-128
	(SEQ. ID NO.: 482)
	SLLNATDIAV	HIV-I GP 160 814-823
	(SEQ. ID NO.: 483)
	VLYRYGSFSV	Pmel gp100 476-485
	(SEQ. ID NO.: 484)
	YIGEVLVSV	Non-filament forming
		class I myosin
		family (HA-2)**
	(SEQ. ID NO.: 485)
	LLFNILGGWV	HCV NS4 192-201
	(SEQ. ID NO.: 486)
	LLVPFVQWFW	HBV env 338-347
	(SEQ. ID NO.: 487)
	ALMPLYACI	HBV pol 642-650
	(SEQ. ID NO.: 488)
	YLVAYQATV	HCV NS3 579-587
	(SEQ. ID NO.: 489)
	TLGIVCPIC	HIPV 16 E7 86-94
	(SEQ. ID NO.: 490)
	YLLPRRGPRL	HCV core protein 34-43
	(SEQ. ID NO.: 491)
	LLPIFFCLWV	HBV env 378-387
	(SEQ. ID NO.: 492)
	YMDDVVLGA	HBV Pol 538-546
	(SEQ. ID NO.: 493)
	GTLGIVCPI	HPV16 E7 85-93
	(SEQ. ID NO.: 494)
	LLALLSCLTI	HCV MP 63-72
	(SEQ. ID NO.: 495)
	MLDLQPETT	HPV 16 E7 12-20
	(SEQ. ID NO.: 496)
	SLMAFTAAV	HCV NS4 174-182
	(SEQ. ID NO.: 497)
	CINGVCWTV	HCV NS3 67-75
	(SEQ. ID NO.: 498)
	VMNILLQYVV	Glutarnic acid decarboxylase
		114-123
	(SEQ. ID NO.: 499)
	ILTVILGVL	Melan A/Mart- 32-40
	(SEQ. ID NO.: 500)
	FLWGPRALV	MAGE-3 271-279
	(SEQ. ID NO.: 501)
	LLCPAGHAV	HCV NS3 163-171
	(SEQ. ID NO.: 502)
	ILDSFDPLV	HCV NSS 239-247
	(SEQ. ID NO.: 503)
	LLLCLIFLL	HBV env 250-258
	(SEQ. ID NO.: 504)
	LIDYQGMLPV	HBV env 260-269
	(SEQ. ID NO.: 505)
	SIVSPFIPLL	HBV env 370-379
	(SEQ. ID NO.: 506)
	FLLTRILTI	HBV env 183-191
	(SEQ. ID NO.: 507)
	HLGNVKYLV	P. faciparum TRAP 3-11
	(SEQ. ID NO.: 508)
	GIAGGLALL	P. faciparum TRAP 500-508
	(SEQ. ID NO.: 509)
	ILAGYGAGV	HCV NS S4A 236-244
	(SEQ. ID NO.: 510)
	GLQDCTMLV	HCV NS5 714-722
	(SEQ. ID NO.: 511)
	TGAPVTYSTY	HCV NS3 281-290
	(SEQ. ID NO.: 512)
	VIYQYMDDLV	HIV-1RT 179-187
	(SEQ. ID NO.: 513)
	VLPDVFIRCV	N-acetylglucosaminyl-
		transferase V Gnt-V intron
	(SEQ. ID NO.: 514)
	VLPDVFIRC	N-acetylglucosaminyl-
		transferase V Gnt-V intron
	(SEQ. ID NO.: 515)
	AVGIGIAVV	Human CD9
	(SEQ. ID NO.: 516)
	LVVLGLLAV	Human glutamyltransferase
	(SEQ. ID NO.: 517)
	ALGLGLLPV	Human G protein
		coupled receptor
	(SEQ. ID NO.: 5 18)
	164-172
	GIGIGVLAA	HSV-I gp C 480-488
	(SEQ. ID NO.: 519)
	GAGIGVAVL	HSV-2 gp C 446-454
	(SEQ. ID NO.: 520)
	IAGIGILAI	Pseudorabies gpGIN
		455-463
	(SEQ. ID NO.: 521)
	LIVIGILIL	Adenovirus 3 E3
		9 kD 30-38
	(SEQ. ID NO.: 522)
	LAGIGLIAA	S. Lincolnensis ImrA
	(SEQ. ID NO.: 523)
	VDGIGILTI	Yeast ysa-1 77-85
	(SEQ. ID NO.: 524)
	GAGIGVLTA	B. polymyxa ,
		βendoxylanase
		149-157
	(SEQ. ID NO.: 525)
	157
	AAGIGHQI	E. coli methionine
		synthase 590-598
	(SEQ. ID NO.: 526)
	QAGIGILLA	E. coli hypothetical
		protein 4-12
	(SEQ. ID NO.: 527)
	KARDPHSGHFV	CDK4wl 22-32
	(SEQ. ID NO.: 528)
	KACDPI-ISGIIFV	CDK4-R24C 22-32
	(SEQ. ID NO.: 529)
	ACDPFISGHFV	CDK4-R24C 23-32
	(SEQ. ID NO.: 530)
	SLYNTVATL	HIV-I gag p17 77-85
	(SEQ. ID NO.: 531)
	ELVSEFSRV	HER-2, m > V
		substituted 971-979
	(SEQ. ID NO.: 532)
	RGPGRAFVTI	HIV-I gp 160 315-329
	(SEQ. ID NO.: 533)
	HMWNFISGI	HCV NS4A 149-157
	(SEQ. ID NO.: 534)
	NLVPMVATVQ	HCMV pp65 495-504
	(SEQ. ID NO.: 535)
	GLHCYEQLV	HPV 6b E7 21-30
	(SEQ. ID NO.: 536)
	PLKQHFQIV	HPV 6b E7 47-55
	(SEQ. ID NO.: 537)
	LLDFVRFMGV	EBNA-6 284-293
	(SEQ. ID NO.: 538)
	AIMEKNIML	Influenza Alaska
		NS 1 122-130
	(SEQ. ID NO.: 539)
	YLKTIQNSL	P. falciparum cp36 CSP
	(SEQ. ID NO.: 540)
	YLNKIQNSL	P. falciparum cp39 CSP
	(SEQ. ID NO.: 541)
	YMLDLQPETT	HPV 16 E7 11-20*
	(SEQ. ID NO.: 542)
	LLMGTLGIV	HPV16 E7 82-90**
	(SEQ. ID NO.: 543)
	TLGIVCPI	HPV 16 E7 86-93
	(SEQ. ID NO.: 544)
	TLTSCNTSV	HIV-1 gp120 197-205
	(SEQ. ID NO.: 545)
	KLPQLCTEL	HPV 16 E6 18-26
	(SEQ. ID NO.: 546)
	TIHDIILEC	HPV 16 E6 29-37
	(SEQ. ID NO.: 547)
	LGIVCPICS	HPV16 E7 87-95
	(SEQ. ID NO.: 548)
	VILGVLLLI	Melan A/Mart-1 35-43
	(SEQ. ID NO.: 549)
	ALMDKSLHV	Melan A/Mart-1 56-64
	(SEQ. ID NO.: 550)
	GILTVILGV	Melan A/Mart-1 31-39
	(SEQ. ID NO.: 551)
T cell	MINAYLDKL	P. Falciparum
epitopes		STARP 523-531
	(SEQ. ID NO.: 552)
	AAGIGILTV	Melan A/Mart- 127-35
	(SEQ. ID NO.: 553)
	FLPSDFFPSV	HBV cAg 18-27
	(SEQ. ID NO.: 554)
Motif	SVRDRLARL	EBNA-3 464-472
unknown
T cell	(SEQ. ID NO.: 555)
epitopes
T cell	AAGIGILTV	Melan A/Mart-1 27-35
epitopes
	(SEQ. ID NO.: 556)
	FAYDGKDYI	Human MHC I-ot 140-148
	(SEQ. ID NO.: 557)
T cell	AAGIGILTV	Melan A/Mart-1 27-35
epitopes
	(SEQ. ID NO.: 558)
	FLPSDFFPSV	HBV cAg 18-27
	(SEQ. ID NO.: 559)
Motif	AAGIGILTV	Meland A/Mart-1 27-35
unknown
T cell	(SEQ. ID NO.: 560)
epitopes
	FLPSDFFPSV	HBV cAg 18-27
	(SEQ. ID NO.: 561)
	AAGIGILTV	Melan A/Mart-1 27-35
	(SEQ. ID NO.: 562)
	ALLAVGATK	Pme117 gp 100 17-25
	(SEQ. ID NO.: 563)
T cell	RLRDLLLIVTR	HIV-1 gp41 768-778
epitopes
	(SEQ. ID NO.: 564)
	QVPLRPMTYK	HIV-1 nef 73-82
	(SEQ. ID NO.: 565)
	TVYYGVPVWK	HIV-1 gp120-36-45
	(SEQ. ID NO.: 566)
	RLRPGGKKK	HIV-1 gag p 17 20-29
	(SEQ. ID NO.: 567)
	ILRGSVAHK	Influenza NP 265-273
	(SEQ. ID NO.: 568)
	RLRAEAGVK	EBNA-3 603-611
	(SEQ. ID NO.: 569)
	RLRDLLLIVTR	HIV-1 gp41 770-780
	(SEQ. ID NO.: 570)
	VYYGVPVWK	HIV-I GP 120 38-46
	(SEQ. ID NO.: 571)
	RVCEKMALY	HCV NS5 575-583
	(SEQ. ID NO.: 572)
Motif	KIFSEVTLK	Unknown; muta melanoma
unknown		peptide ted (p I 83L)
		175-183
T cell	(SEQ. ID NO.: 573)
epitope
	YVNVNMGLK*	HBV cAg 88-96
	(SEQ. ID NO.: 574)
T cell	IVTDFSVIK	EBNA-4 416-424
epitopes
	(SEQ. ID NO.: 575)
	ELNEALELK	P53 343-351
	(SEQ. ID NO.: 576)
	VPLRPMTYK	HIV-1 NEF 74-82
	(SEQ. ID NO.: 577)
	AIFQSSMTK	HIV-I gag p24 325-333
	(SEQ. ID NO.: 578)
	QVPLRPMTYK	HIV-1 nef 73-82
	(SEQ. ID NO.: 579)
	TINYTIFK HCV	NSI 238-246
	(SEQ. ID NO.: 580)
	AAVDLSHFLKEK	HIV-1 nef 83-94
	(SEQ. ID NO.: 581)
	ACQGVGGPGGHK	HIV-1 II 1B p24 349-359
	(SEQ. ID NO.: 581)
HLA-A24	SYLDSGIHF*	β-catenin, mutated
		(proto-onocogen)
		29-37
	(SEQ. ID NO.: 582)
T cell	RYLKDQQLL	HIV GP 41 583-591
epitopes
	(SEQ. ID NO.: 583)
	AYGLDFYIL	P15 melanoma Ag 10-18
	(SEQ. ID NO.: 584)
	AFLPWHRLFL	Tyrosinase 206-215
	(SEQ. ID NO.: 585)
	AFLPWHRLF	Tyrosinase 206-214
	(SEQ. ID NO.: 586)
	RYSIFFDY	Ebna-3 246-253
	(SEQ. ID NO.: 587)
T cell	ETINEEAAEW	HIV-1 gag p24 203-212
epitope
	(SEQ. ID NO.: 588)
T cell	STLPETTVVRR	HBV cAg 141-151
epitopes
	(SEQ. ID NO.: 589)
	MSLQRQFLR	ORF 3P-gp75
		294-321 (bp)
	(SEQ. ID NO.: 590)
	LLPGGRPYR	TRP (tyrosinase rel.)
		197-205
	(SEQ. ID NO.: 591)
T cell	IVGLNKIVR	HIV gag p24
epitope		267-267-275
	(SEQ. ID NO.: 592)
	AAGIGILTV	Melan A/Mart-127 35
	(SEQ. ID NO.: 593)

[0080] Table 3 sets forth additional antigens useful in the invention that are available from the Ludwig Cancer Institute. The Table refers to patents in which the identified antigens can be found and as such are incorporated herein by reference. TRA refers to the tumor-related antigen and the LUD No. refers to the Ludwig Institute number.

TABLE 3
	LUD		Date Patent	Peptide
TRA	No.	Patent No.	Issued	(Antigen)	HLA
MAGE-4	5293	5,405,940	11 Apr. 1995	EVDPASNTY	HLA-A1
				(SEQ. ID NO.: 532)
MAGE-41	5293	5,405,940	11 Apr. 1995	EVDPTSNTY	HLA-A I
				(SEQ ID NO: 533)
MAGE-5	5293	5,405,940	11 Apr. 1995	EADPTSNTY	HLA-A I
				(SEQ ID NO: 534)
MAGE-51	5293	5,405,940	11 Apr. 1995	EADPTSNTY	HLA-A I
				(SEQ ID NO: 534)
MAGE-6	5294	5,405,940	11 Apr. 1995	EVDPIGHVY	HLA-A1
				(SEQ ID NO: 535)
	5299.2	5,487,974	30 Jan. 1996	MLLAVLYCLL	HLA-A2
				(SEQ ID NO: 536)
	5360	5,530,096	25 Jun. 1996	MLLAVLYCL	HLA-B44
				(SEQ ID NO: 537)
Tyrosinase	5360.1	5,519,117	21 May 1996	SEIWRDIDFA	HLA-B44
				(SEQ ID NO: 538)
				SEIWRDIDF
				(SEQ ID NO: 539)
Tyrosinase	5431	5,774,316	28 Apr. 1998	XEIWRDIDF	HLA-B44
				(SEQ ID NO: 540)
MAGE-2	5340	5,554,724	10 Sep. 1996	STLVEVTLGEV	HLA-A2
				(SEQ ID NO: 541)
				LVEVTLGEV
				(SEQ ID NO: 542)
				VIFSKASEYL
				(SEQ ID NO: 543)
				IIVLAIIAI
				(SEQ ID NO: 544)
				KIWEELSMLEV
				(SEQ ID NO: 545)
				LIETSYVKV
				(SEQ ID NO: 546)
	5327	5,585,461	17 Dec. 1996	FLWGPRALV	HLA-A2
				(SEQ ID NO: 547)
				TLVEVTLGEV
				(SEQ ID NO: 548)
				ALVETSYVKV
				(SEQ ID NO: 549)
MAGE-3	5344	5,554,506	10 Sep. 1996	KIWEELSVL	HLA-A2
				(SEQ ID NO: 550)
MAGE-3	5393	5,405,940	11 Apr. 1995	EVDPIGHLY	HLA-A1
				(SEQ ID NO: 551)
MAGE	5293	5,405,940	11 Apr. 1995	EXDX5Y	HLA-A1
				(SEQ. ID NO.: 552)
				(but not EADPTGHSY)
				(SEQ. ID NO.: 553)
				E (A/V) D X5 Y
				(SEQ. ID NO.: 554)
				E (A/V) D P X4 Y
				(SEQ. ID NO.: 555)
				E (A/V) D P (I/A/T)
				X3 Y
				(SEQ. ID NO.: 556)
				E (A/V) D P (I/A/T)
				(G/S) X2 Y
				(SEQ. ID NO.: 557)
				E (A/V) D P (I/A/T)
				(G/S) (H/N) X Y
				E (A/V) DP (I/A/T)
				(G/S) (H/N)
				(L/T/V) Y
				(SEQ. 11) NO.: 559)
MAGE-1	5361	5,558.995	24 Sep. 1996	ELHSAYGEPRKLLTQD	HLA-C
				(SEQ ID NO: 560)	Clone 10
				EHSAYGEPRKLL
				(SEQ ID NO: 561)
				SAYGEPRKL
				(SEQ ID NO: 562)
MAGE-1	5253.4	TBA	TBA	EADPTGHSY	HLA-A I
				(SEQ ID NO: 563)
BAGE	5310.1	TBA	TBA	MAARAVFLALSAQLLQARLMKE	HLA-C
				(SEQ ID NO: 564)	Clone 10
				MAARAVFLALSAQLLQ	HLA-C
				(SEQ ID NO: 565)	Clone 10
				AARAVFLAL	HLA-C
				(SEQ ID NO: 566)	Clone 10
GAGE	5323.2	5,648,226	15 Jul. 1997	YRPRPRRY	HLA-CW6
				(SEQ. ID NO.: 567)

[0081]

TABLE 4
		AA	MHC	T cell epitope MHC
Source	Protein	Position	molecules	ligand (Antigen)	Ref.
synthetic	synthetic	synthetic	HLA-A2	ALFAAAAAV	Parker, et al., “Scheme for ranking
peptides	peptides	peptides			potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLDKGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGFGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGAGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGEGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGFGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGGGL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGVGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGVGKV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFKGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLGGGGFGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLLGGGVGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLYGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GMFGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GMFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GQFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GVFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KLFGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KLFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	AILGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GAIGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GALGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GELGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIAGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIEGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILAFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGAVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGEVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILFGAFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFEFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFKFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVATL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVETL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVFAL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVFEL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVFKL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVFTA	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVFVL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGFVKTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILGKVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILKFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GILPFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIVGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GKLGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLLGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GQLGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KALGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KILGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KILGKVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	AILLGVFML	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	AIYKRWIIL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ALFFFDIDL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ATVELLSEL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	CLFGYPVYV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	FIFPNYTIV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	IISLWDSQL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ILASLFAAV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ILESLFAAV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KLGEFFNQM	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KLGEFYNQM	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	LLFGYPVYV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	LLWKGEGAV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	LMFGYPVYV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	LNFGYPVYV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	LQFGYPVYV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	NIVAHTFKV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	NLPMVATV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	QMLLAIARL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	QMWQARLTV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	RLLQTGIHV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	RLVNGSLAL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	SLYNTVATL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	TLNAWVKVV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	WLYRETCNL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	YLFKRMIDL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GAFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GAFGGVGGY	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GEFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GGFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIFGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIGGFGGGL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIGGGGGGL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLDGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLDGKGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLDKKGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGFGF	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGFGG	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGFGN	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGFGS	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGGGI	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGGGM	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGGGT	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGGGY	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLGFGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLGGFGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLGGGFGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLGGGGGFV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLGGGGGGY	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLGGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLLGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLPGGGGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GNFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GSFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GTFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	AGNSAYEYV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFPGQFAY	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	HILLGVFML	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ILESLFRAV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KKKYKLKHI	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	MLASIDLKY	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	MLERELVRK	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KLFGFVFTV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ILDKKVEKV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ILKEPVHGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ALFAAAAAY	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIGFGGGGL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GKFGGVGGV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GLFGGGGGK	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	EILGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GIKGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	GQLGFVFTK	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	ILGFVFTLT	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KILGFVFTK	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KKLGFVFTL	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	KLFEKVYNY	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
			″	LRFGYPVYV	Parker, et al., “Scheme for ranking
					potential HLA-A2 binding peptides based
					on independent binding of individual peptide
					side-chains,” J. Immunol. 152:163-175
Human	HSP60	140-148	HLA-B27	IRRGVMLAV	Rammensee et al. 1997
					160
″	″	369-377	″	KRIQEIIEQ	Rammensee et al. 1997
					160
″	″	469-477	″	KRTLKIPAM	Rammensee et al. 1997
					160
Yersinia	HSP60	35-43	″	GRNVVLDKS	Rammensee et al. 1997
					160
″	″	117-125	″	KRGIDKAVI	Rammensee et al. 1997
					160
″	″	420-428	″	IRAASAITA	Rammensee et al. 1997
					160
″	HSP60	284-292	HLA-	RRKAMFEDI	169
			B*2705
P. falciparum	LSA-1	1850-	HLA-	KPKDELDY	170
		1857	B3501
Influenza		379-387	HLA-	LELRSRYWA	183
NP			B*4402
	Tum-P35B	4-13	HLA-Dd	GPPHSNNFGY	230
Rotavirus	VP7	33-40		IIYRFLLI	262
	OGDH	104-112	H2-Ld	QLSPYPFDL	253
	(F108Y)
	TRP-2	181-188	p287	VYDFFVWL	284
	DEAD box	547-554	p287	SNFVFAGI	283
	p 68
	Vector		p287	SVVEFSSL	260
	“artefact”
	Epiope		p287	AHYLFRNL	278
	mimic of
	tumor Ag
	Epitope		″	THYLFRNL	″
	mimic of
	H-3
	Epitope		″	LIVIYNTL	279
	mimic of
	H-3
	miHAg″
			″	LIYEFNTL	″
			″	IPYIYNTL	″
			″	IIYIYHRL	″
			″	LIYIFNTL	″
	HBV cAg	93-100	″	MGLKFRQL	280
Human	autoantigen	51-58	″	IMIKFRNRL	281
	LA
Mouse	UTY		H2Db	WMHHNMDLI	303
	protein
Mouse	p53	232-240	″	KYMCNSSCM	302
MURINE	MDM2	441-449	″	GRPKNGCIV	277
	Epitope		″	AQHPNAELL	278
	mimic of
	natural
	MuLV	75-83	″	CCLCLTVFL	301
	gag75K
P. falciparum	CSP	375-383	p290	YENDIEKK	315
″	″	371-379	″	DELDYENDI	315
HIV	−1RT	206-214	″	TEMEKEGKI	316
Rabies	NS	197-205		VEAEIAHQI	309, 310
Influenza A	NS1	152-160	″	EEGAIVGEI	304
Murine	SMCY		p291	TENSGKDI	317
	MHC class	3-11	p293	AMAPRTLLL	318
	1 leader
	ND1 alpha	1-12	p293	FFINILTLLVP	323
	ND Beta	1-12	p293	FFINILTLLVP	323
	ND alpha	1-17	″	FFINILTLLVPILIAM	324
	ND Beta	1-17	″	FFINALTLLVPILIAM	″
	COI	1-6	″	FINRW	325
	mitochondrial
L.	LemA	1-6	″	IGWII	326
monocytogenes
	SIV gag	179-190	Mamu-	EGCTPYDINQML	334
	p11C		A*01
	MAGE-3		HLA-A2	ALSRKVAEL	5,554,506
			″	IMPKAGLLI	″
			″	KIWEELSVL	″
			″	ALVETSYVKV	″
			″	Thr Leu Val Glu Val	″
				Thr Leu Gly Glu Val
			″	Ala Leu Ser Arg Lys	″
				Val Ala Glu Leu
			″	Ile Met Pro Lys Ala	″
				Gly Leu Leu Ile
			″	Lys Ile Trp Glu Glu	″
				Leu Ser Val Leu
			″	Ala Leu Val Glu Thr	″
				Ser Tyr Val Lys Val
	peptides		HLA-A2	Lys Gly Ile Leu Gly	5,989,565
	which bind			Phe Val Phe Thr Leu
	to MHCs			Thr Val
			″	Gly Ile Ile Gly Phe	″
				Val Phe Thr Ile
			″	Gly Ile Ile Gly Phe	″
				Val Phe Thr Leu
			″	Gly Ile Leu Gly Phe	″
				Val Phe Thr Leu
			″	Gly Leu Leu Gly Phe	″
				Val Phe Thr Leu
			″	XXTVXXGVX, X =	″
				Leu or Ile (6-37)
			″	Ile Leu Thr Val Ile	″
				Leu Gly Val Leu
			″	Tyr Leu Glu Pro Gly	″
				Pro Val Thr Ala
			″	Gln Val Pro Leu Arg	″
				Pro Met Thr Tyr Lys
			″	Asp Gly Leu Ala Pro	″
				Pro Gln His Leu Ile
				Arg
			″	Leu Leu Gly Arg Asn	″
				Ser Phe Glu Val
	Peptides		HLA-C	Glu His Ser Ala Tyr	5,558,995
	from		clone 10	Gly Glu Pro Arg Lys
	MAGE-1			Leu Leu Thr Gln Asp
				Leu
			HLA-C	Glu His Ser Ala Tyr	″
			clone 10	Gly Glu Pro Arg Lys
				Leu Leu
			HLA-C	Ser Ala Tyr Gly Glu	″
			clone 10	Pro Arg Lys Leu
	GAGE		HLA-Cw6	Tyr Arg Pro Arg Pro	5,648,226
				Arg Arg Tyr
			″	Thr Tyr Arg Pro Arg	″
				Pro Arg Arg Tyr
			″	Tyr Arg Pro Arg Pro	″
				Arg Arg Tyr Val
			″	Thr Tyr Arg Pro Arg	″
				Pro Arg Arg Tyr Val
			″	Arg Pro Arg Pro Arg	″
				Arg Tyr Val Glu
			″	Met Ser Trp Arg Gly	″
				Arg Ser Thr Tyr Arg
				Pro Arg Pro Arg Arg
			″	Thr Tyr Arg Pro Arg	″
				Pro Arg Arg Tyr Val
				Glu Pro Pro Glu Met
				Ile
	MAGE		HLA-A1,	Isolated nonapeptide	5,405,940
			primarily	having Glu at its N
				terminal, Tyr at its C-
				terminal, and Asp at
				the third residue from
				its N terminal, with
				the proviso that said
				isolated nonapeptide
				is not Glu Ala Asp
				Pro Thr Gly His Ser
				Tyr (SEQ ID NO: 1),
				and wherein said
				isolated nonapeptide
				binds to a human
				leukocyte antigen
				molecule on a cell to
				form a complex, said
				complex provoking
				lysis of said cell by a
				cytolytic T cell
				specific to said
				complex
			HLA-A1,	Glu Val Val Pro Ile	″
			primarily	Ser His Leu Tyr
			HLA-A1,	Glu Val Val Arg Ile	″
			primarily	Gly His Leu Tyr
			HLA-A1,	Glu Val Asp Pro Ile	″
			primarily	Gly His Leu Tyr
			HLA-A1,	Glu Val Asp Pro Ala	″
			primarily	Ser Asn Thr Tyr
			HLA-A1,	Glu Val Asp Pro Thr	″
			primarily	Ser Asn Thr Tyr
			HLA-A1,	Glu Ala Asp Pro Thr	″
			primarily	Ser Asn Thr Tyr
			HLA-A1,	Glu Val Asp Pro Ile	″
			primarily	Gly His Val Tyr
			HLA-A1,	GAAGTGGTCCCC	″
			primarily	ATCAGCCACTTGTAC
			HLA-A1,	GAAGTGGTCCGC	″
			primarily	ATCGGCCACTTGTAC
			HLA-A1,	GAAGTGGACCCC	″
			primarily	ATCGGCCACTTGTAC
			HLA-A1,	GAAGTGGACCCC	″
			primarily	GCCAGCAACACCTAC
			HLA-A1,	GAAGTGGACCCC	″
			primarily	ACCAGCAACACCTAC
			HLA-A1,	GAAGCGGACCCC	″
			primarily	ACCAGCAACACCTAC
			HLA-A1,	GAAGCGGACCCC	″
			primarily	ACCAGCAACACCTAC
			HLA-A1,	GAAGTGGACCCC	″
			primarily	ATCGGCCACGTGTAC
			HLA-A1,	Glu Ala Asp Pro Thr	″
			primarily	Gly His Ser
			HLA-A1,	Ala Asp Pro Trp Gly	″
			primarily	His Ser Tyr
	MAGE peptides		HLA-A2	Ser Thr Leu Val Glu	5,554,724
				Val Thr Leu Gly Glu
				Val
	″		″	Leu Val Glu Val Thr	″
				Leu Gly Glu Val
	″		″	Lys Met Val Glu Leu	″
				Val His Phe Leu
	″		″	Val Ile Phe Ser Lys	″
				Ala Ser Glu Tyr Leu
	″		″	Tyr Leu Gln Leu Val	″
				Phe Gly Ile Glu Val
	″		″	Gln Leu Val Phe Gly	″
				Ile Glu Val Val
	″		″	Gln Leu Val Phe Gly	″
				Ile Glu Val Val Glu
				Val
	″		″	Ile Ile Val Leu Ala	″
				Ile Ile Ala Ile
	″		″	Lys Ile Trp Glu Glu	″
				Leu Ser Met Leu Glu
				Val
	″		″	Ala Leu Ile Glu Thr	″
				Ser Tyr Val Lys Val
	″		″	Leu Ile Glu Thr Ser	″
				Tyr Val Lys Val
	″		″	Gly Leu Glu Ala Arg	″
				Gly Glu Ala Leu Gly
				Leu
	″		″	Gly Leu Glu Ala Arg	″
				Gly Glu Ala Leu
	″		″	Ala Leu Gly Leu Val	″
				Gly Ala Gln Ala
	″		″	Gly Leu Val Gly Ala	″
				Gln Ala Pro Ala
	″		″	Asp Leu Glu Ser Glu	″
				Phe Gln Ala Ala
	″		″	Asp Leu Glu Ser Glu	″
				Phe Gln Ala Ala Ile
	″		″	Ala Ile Ser Arg Lys	″
				Met Val Glu Leu Val
	″		″	Ala Ile Ser Arg Lys	″
				Met Val Glu Leu
	″		″	Lys Met Val Glu Leu	″
				Val His Phe Leu Leu
	″		″	Lys Met Val Glu Leu	″
				Val His Phe Leu Leu
				Leu
	″		″	Leu Leu Leu Lys Tyr	″
				Arg Ala Arg Glu Pro
				Val
	″		″	Leu Leu Lys Tyr Arg	″
				Ala Arg Glu Pro Val
	″		″	Val Leu Arg Asn Cys	″
				Gln Asp Phe Phe Pro
				Val
	″		″	Tyr Leu Gln Leu Val	″
				Phe Gly Ile Glu Val
				Val
	″		″	Gly Ile Glu Val Val	″
				Glu Val Val Pro Ile
	″		″	Pro Ile Ser His Leu	″
				Tyr Ile Leu Val
	″		″	His Leu Tyr Ile Leu	″
				Val Thr Cys Leu
	″		″	His Leu Tyr Ile Leu	″
				Val Thr Cys Leu Gly
				Leu
	″		″	Tyr Ile Leu Val Thr	″
				Cys Leu Gly Leu
	″		″	Cys Leu Gly Leu Ser	″
				Tyr Asp Gly Leu
	″		″	Cys Leu Gly Leu Ser	″
				Tyr Asp Gly Leu Leu
	″		″	Val Met Pro Lys Thr	″
				Gly Leu Leu Ile
	″		″	Val Met Pro Lys Thr	″
				Gly Leu Leu Ile Ile
	″		″	Val Met Pro Lys Thr	″
				Gly Leu eu Ile Ile
				Val
	″		″	Gly Leu Leu Ile Ile	″
				Val Leu Ala Ile
	″		″	Gly Leu Leu Ile Ile	″
				Val Leu Ala Ile Ile
	″		″	Gly Leu Leu Ile Ile	″
				Val Leu Ala Ile Ile
				Ala
	″		″	Leu Leu Ile Ile Val	″
				Leu Ala Ile Ile
	″		″	Leu Leu Ile Ile Val	″
				Leu Ala Ile Ile Ala
	″		″	Leu Leu Ile Ile Val	″
				Leu Ala Ile Ile Ala
				Ile
	″		″	Leu Ile Ile Val Leu	″
				Ala Ile Ile Ala
	″		″	Leu Ile Ile Val Leu	″
				Ala Ile Ile Ala Ile
	″		″	Ile Ile Ala Ile Glu	″
				Gly Asp Cys Ala
	″		″	Lys Ile Trp Glu Glu	″
				Leu Ser Met Leu
	″		″	Leu Met Gln Asp Leu	″
				Val Gln Glu Asn Tyr
				Leu
	″		″	Phe Leu Trp Gly Pro	″
				Arg Ala Leu Ile
	″		″	Leu Ile Glu Thr Ser	″
				Tyr Val Lys Val
	″		″	Ala Leu Ile Glu Thr	″
				Ser Tyr Val Lys Val
				Leu
	″		″	Thr Leu Lys Ile Gly	″
				Gly Glu Pro His Ile
	″		″	His Ile Ser Tyr Pro	″
				Pro Leu His Glu Arg
				Ala
	″		″	Gln Thr Ala Ser Ser	″
				Ser Ser Thr Leu
	″		″	Gln Thr Ala Ser Ser	″
				Ser Ser Thr Leu Val
	″		″	Val Thr Leu Gly Glu	″
				Val Pro Ala Ala
	″		″	Val Thr Lys Ala Glu	″
				Met Leu Glu Ser Val
	″		″	Val Thr Lys Ala Glu	″
				Met Leu Glu Ser Val
				Leu
	″		″	Val Thr Cys Leu Gly	″
				Leu Ser Tyr Asp Gly
				Leu
	″		″	Lys Thr Gly Leu Leu	″
				Ile Ile Val Leu
	″		″	Lys Thr Gly Leu Leu	″
				Ile Ile Val Leu Ala
	″		″	Lys Thr Gly Leu Leu	″
				Ile Ile Val Leu Ala
				Ile
	″		″	His Thr Leu Lys Ile	″
				Gly Gly Glu Pro His
				Ile
	″		″	Met Leu Asp Leu Gln	″
				Pro Glu Thr Thr
	Mage-3 peptides		HLA-A2	Gly Leu Glu Ala Arg	5,585,461
				Gly Glu Ala Leu
	″		″	Ala Leu Ser Arg Lys	″
				Val Ala Glu Leu
	″		″	Phe Leu Trp Gly Pro	″
				Arg Ala Leu Val
	″		″	Thr Leu Val Glu Val	″
				Thr Leu Gly Glu Val
	″		″	Ala Leu Ser Arg Lys	″
				Val Ala Glu Leu Val
	″		″	Ala Leu Val Glu Thr	″
				Ser Tyr Val Lys Val
	Tyrosinase		HLA-A2	Tyr Met Asn Gly Thr	5,487,974
				Met Ser Gln Val
	″		″	Met Leu Leu Ala Val	″
				Leu Tyr Cys Leu Leu
	Tyrosinase		HLA-A2	Met Leu Leu Ala Val	5,530,096
				Leu Tyr Cys Leu
	″		″	Leu Leu Ala Val Leu	″
				Tyr Cys Leu Leu
	Tyrosinase		HLA-A2	Ser Glu Ile Trp Arg	5,519,117
			and HLA-B44	Asp Ile Asp Phe Ala
				His Glu Ala
	″		HLA-A2	Ser Glu Ile Trp Arg	″
			and HLA-B44	Asp Ile Asp Phe
	″		HLA-A2	Glu Glu Asn Leu Leu	″
			and HLA-B44	Asp Phe Val Arg Phe
	Melan			EAAGIGILTV	Jäger, E. et al. Granulocyte-
	A/MART-1				macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
	Tyrosinase			MLLAVLYCL	Jäger, E. et al. Granulocyte-
					macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
	″			YMDGTMSQV	Jäger, E. et al. Granulocyte-
					macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
	gp100/Pme 117			YLEPGPVTA	Jäger, E. et al. Granulocyte-
					macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
	″			LLDGTATLRL	Jäger, E. et al. Granulocyte-
					macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
	Influenza			GILGFVFTL	Jäger, E. et al. Granulocyte-
	matrix				macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
	MAGE-1			EADPTGHSY	Jäger, E. et al. Granulocyte-
					macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
					Jäger, E. et al. Granulocyte-
					macrophage-colony-stimulating Factor
					Enhances Immune Responses To
					Melanoma-′associated Peptides in vivo
					Int. J Cancer 67, 54-62 (1996)
	MAGE-1		HLA-A1	EADPTGHSY	DIRECTLY FROM
					DAVID'S LIST
	BAGE		HLA-C	MAARAVFLALSA	DIRECTLY FROM
				QLLQARLMKE	DAVID'S LIST
	″		″	MAARAVFLALSA	DIRECTLY FROM
				QLLQ	DAVID'S LIST
	″		″	AARAVFLAL	DIRECTLY FROM
					DAVID'S LIST
Influenza	PR8 NP	147-154	Kd	IYQRIRALV	Falk et al., Allele-specific motifs
					revealed by sequencing of self-
					peptides eluted from MHC molecules
SELF	P815		″	SYFPEITHI	Falk et al., Allele-specific motifs
PEPTIDE					revealed by sequencing of self-
					peptides eluted from MHC molecules
Influenza	Jap HA		″	IYATVAGSL	Falk et al., Allele-specific motifs
	523-549				revealed by sequencing of self-
					peptides eluted from MHC molecules
″	Jap HA		″	VYQILAIYA	Falk et al., Allele-specific motifs
	523-549				revealed by sequencing of self-
					peptides eluted from MHC molecules
″	Jap HA		″	IYSTVASSL	Falk et al., Allele-specific motifs
	523-549				revealed by sequencing of self-
					peptides eluted from MHC molecules
″	JAP HA		″	LYQNVGTYV	Falk et al., Allele-specific motifs
	202-221				revealed by sequencing of self-
					peptides eluted from MHC molecules
	HLA-A24		″	RYLENQKRT	Falk et al., Allele-specific motifs
					revealed by sequencing of self-
					peptides eluted from MHC molecules
	HLA-Cw3		″	RYLKNGKET	Falk et al., Allele-specific motifs
					revealed by sequencing of self-
					peptides eluted from MHC molecules
	P815		″	KYQAVTTTL	Falk et al., Allele-specific motifs
					revealed by sequencing of self-
					peptides eluted from MHC molecules
Plasmodium	CSP		″	SYIPSAEKI	Falk et al., Allele-specific motifs
berghen					revealed by sequencing of self-
					peptides eluted from MHC molecules
Plasmodium	CSP		″	SYVPSAFQI	Falk et al., Allele-specific motifs
yoelli					revealed by sequencing of self-
					peptides eluted from MHC molecules
Vesicular	NP 52-59		Kb	RGYVYQGL	Falk et al., Allele-specific motifs
stomatitis					revealed by sequencing of self-
viruse					peptides eluted from MHC molecules
Ovalbumin			″	SIINFEKL	Falk et al., Allele-specific motifs
					revealed by sequencing of self-
					peptides eluted from MHC molecules
Sandal	NP 321-		″	APGNYPAL	Falk et al., Allele-specific motifs
Virus	332				revealed by sequencing of self-
					peptides eluted from MHC molecules
				VPYGSFKHV	Morel et al., Processing of some
					antigens by the standard proteasome
					but not by the immunoproteasome
					results in poor presentation by
					dendritic cells, Immunity, vol.
					12:107-117, 2000.
MOTIFS
influenza	PR8 NP		Kd	TYQRTRALV	5,747,269
			restricted
			peptide
			motif
self peptide	P815		Kd	SYFPEITHI	″
			restricted
			peptide
			motif
influenza	JAP HA		Kd	IYATVAGSL	″
			restricted
			peptide
			motif
influenza	JAP HA		Kd	VYQILAIYA	″
			restricted
			peptide
			motif
influenza	PR8 HA		Kd	IYSTVASSL	″
			restricted
			peptide
			motif
influenza	JAP HA		Kd	LYQNVGTYV	″
			restricted
			peptide
			motif
			HLA-A24	RYLENGKETL	″
			HLA-Cw3	RYLKNGKETL	″
	P815		″	KYQAVTTTL	″
	tumour
	antigen
Plasmodium	CSP		″	SYIPSAEKI	″
berghei
Plasmodium	CSP		″	SYVPSAEQI	″
yoeli
influenza	NP		Db-	ASNENMETM	″
			restricted
			peptide
			motif
adenovirus	E1A		Db-	SGPSNTPPEI	″
			restricted
			peptide
			motif
lymphocytic			Db-	SGVENPGGYCL	″
choriomeningitis			restricted
			peptide
			motif
simian	40 T		Db-	SAINNY . . .	″
virus			restricted
			peptide
			motif
HIV	reverse		HLA-A2.1-	ILKEPVHGV	″
	transcriptase		restricted
			peptide
			motif
	influenza		HLA-A2.1-	GILGFVFTL	″
	matrix		restricted
	protein		peptide
			motif
influenza	influenza		HLA-A2.1-	ILGFVFTLTV	″
	matrix		restricted
	protein		peptide
			motif
HIV	Gag			FLQSRPEPT	″
	protein
HIV	Gag			AMQMLKE . . .	″
	protein
HIV	Gag			PLAPGQMRE	″
	protein
HIV	Gag			QMKDCTERQ	″
	protein
			HLA-A*0205-	VYGVIQK	″
			restricted
			peptide
			motif

[0082]

TABLE 5
VSV-NP peptide (49-62)
LCMV-NP peptide (118-132)
LCMV glycoprotein peptide. 33-41
ISNQLTLDSNTKYFHKLN
ISNQLTLDSNTKYFHKL
ISNQLTLDSNTKYFHK
VDTFLEDVKNLYHSEA
KPRAIVVDPVHGFMY
KQTISPDYRNMI
Y[DFIMDPKEKDKV
NIQLINDQEVARFD
LLSFVRDLNQYRADI
LPKPPKPVSKMRMATPL
LPKPPKPVSKMRMATPLLMQALP
LPKPPKPVSKMRMATPLLMQALPM
PKPPKPVSKMRMATPL
PKPPKPVSKMRMATPLLMQA
KPPKPVSKMRMATPLLMQ
KPPKPVSKMRMATPLLMQALPM
VDDTQFVRFDSDAASQ
ATKYGNMTEDHVMHLLQNA
VFLLLLADKVPETSLS
LNKILLDEQAQWK
GPPKLDIRKEEKQIMIDIFH
GPPKLDIRKEEKQIMIDIFHP
GFKAIRPDKKSNPIIRTV
YANILLDRRVPQTDMTF
NLFLKSDGRIKYTLNKNSLK
IPDNLFLKSDGRIKYTLNKN
IPDNLFLKSDGRIKYTLNK
IPDNLFLKSDGRIKYTLN
IPDNLFLKSDGRIKYTL
NLFLKSDGRIKYTLNK
NLFLKSDGRIKYTLN
VTTLNSDLKYNALDLTN
VGSDWRFLRGYHQYA

[0083] Still further embodiments are directed to methods, uses, therapies and compositions related to epitopes with specificity for MHC, including, for example, those listed in Tables 6-10. Other embodiments include one or more of the MHCs listed in Tables 6-10, including combinations of the same, while other embodiments specifically exclude any one or more of the MHCs or combinations thereof. Tables 8-10 include frequencies for the listed HLA antigens.

TABLE 6
Class I MHC Molecules
Class I
Human
HLA-A1
HLA-A*0101
HLA-A*0201
HLA-A*0202
HLA-A*0203
HLA-A*0204
HLA-A*0205
HLA-A*0206
HLA-A*0207
HLA-A*0209
HLA-A*0214
HLA-A3
HLA-A*0301
HLA-A*1101
HLA-A23
HLA-A24
HLA-A25
HLA-A*2902
HLA-A*3101
HLA-A*3302
HLA-A*6801
HLA-A*6901
HLA-B7
HLA-B*0702
HLA-B*0703
HLA-B*0704
HLA-B*0705
HLA-B8
HLA-B13
HLA-B14
HLA-B*1501 (B62)
HLA-B17
HLA-B18
HLA-B22
HLA-B27
HLA-B*2702
HLA-B*2704
HLA-B*2705
HLA-B*2709
HLA-B35
HLA-B*3501
HLA-B*3502
HLA-B*3701
HLA-B*3801
HLA-B*39011
HLA-B*3902
HLA-B40
HLA-B*40012 (B60)
HLA-B*4006 (B61)
HLA-B44
HLA-B*4402
HLA-B*4403
HLA-B*4501
HLA-B*4601
HLA-B51
HLA-B*5101
HLA-B*5102
HLA-B*5103
HLA-B*5201
HLA-B*5301
HLA-B*5401
HLA-B*5501
HLA-B*5502
HLA-B*5601
HLA-B*5801
HLA-B*6701
HLA-B*7301
HLA-B*7801
HLA-Cw*0102
HLA-Cw*0301
HLA-Cw*0304
HLA-Cw*0401
HLA-Cw*0601
HLA-Cw*0602
HLA-Cw*0702
HLA-Cw8
HLA-Cw*1601M
HLA-G
Murine
H2-Kd
H2-Dd
H2-Ld
H2-Kb
H2-Db
H2-Kk
H2-Kkm1
Qa-1a
Qa-2
H2-M3
Rat
RT1.Aa
RT1.A1
Bovine
Bota-A11
Bota-A20
Chicken
B-F4
B-F12
B-F15
B-F19
Chimpanzee
Patr-A*04
Patr-A*11
Patr-B*01
Patr-B*13
Patr-B*16
Baboon
Papa-A*06
Macaque
Mamu-A*01
Swine
SLA (haplotype d/d)
Virus homolog
hCMV class I homolog UL18

[0084]

TABLE 7
Class I MHC Molecules
Class I
Human
HLA-A1
HLA-A*0101
HLA-A*0201
HLA-A*0202
HLA-A*0204
HLA-A*0205
HLA-A*0206
HLA-A*0207
HLA-A*0214
HLA-A3
HLA-A*1101
HLA-A24
HLA-A*2902
HLA-A*3101
HLA-A*3302
HLA-A*6801
HLA-A*6901
HLA-B7
HLA-B*0702
HLA-B*0703
HLA-B*0704
HLA-B*0705
HLA-B8
HLA-B14
HLA-B*1501 (B62)
HLA-B27
HLA-B*2702
HLA-B*2705
HLA-B35
HLA-B*3501
HLA-B*3502
HLA-B*3701
HLA-B*3801
HLA-B*39011
HLA-B*3902
HLA-B40
HLA-B*40012 (B60)
HLA-B*4006 (B61)
HLA-B44
HLA-B*4402
HLA-B*4403
HLA-B*4601
HLA-B51
HLA-B*5101
HLA-B*5102
HLA-B*5103
HLA-B*5201
HLA-B*5301
HLA-B*5401
HLA-B*5501
HLA-B*5502
HLA-B*5601
HLA-B*5801
HLA-B*6701
HLA-B*7301
HLA-B*7801
HLA-Cw*0102
HLA-Cw*0301
HLA-Cw*0304
HLA-Cw*0401
HLA-Cw*0601
HLA-Cw*0602
HLA-Cw*0702
HLA-G
Murine
H2-Kd
H2-Dd
H2-Ld
H2-Kb
H2-Db
H2-Kk
H2-Kkml
Qa-2
Rat
RT1.Aa
RT1.A1
Bovine
Bota-A11
Bota-A20
Chicken
B-F4
B-F12
B-F15
B-F19
Virus homolog
hCMV class I homolog UL18

[0085]

TABLE 8
Estimated gene frequencies of HLA-A antigens
	CAU	AFR	ASI	LAT	NAT
Antigen	Gfa	SEb	Gf	SE	Gf	SE	Gf	SE	Gf	SE
A1	15.1843	0.0489	5.7256	0.0771	4.4818	0.0846	7.4007	0.0978	12.0316	0.2533
A2	28.6535	0.0619	18.8849	0.1317	24.6352	0.1794	28.1198	0.1700	29.3408	0.3585
A3	13.3890	0.0463	8.4406	0.0925	2.6454	0.0655	8.0789	0.1019	11.0293	0.2437
A28	4.4652	0.0280	9.9269	0.0997	1.7657	0.0537	8.9446	0.1067	5.3856	0.1750
A36	0.0221	0.0020	1.8836	0.0448	0.0148	0.0049	0.1584	0.0148	0.1545	0.0303
A23	1.8287	0.0181	10.2086	0.1010	0.3256	0.0231	2.9269	0.0628	1.9903	0.1080
A24	9.3251	0.0395	2.9668	0.0560	22.0391	0.1722	13.2610	0.1271	12.6613	0.2590
A9 unsplit	0.0809	0.0038	0.0367	0.0063	0.0858	0.0119	0.0537	0.0086	0.0356	0.0145
A9 total	11.2347	0.0429	13.2121	0.1128	22.4505	0.1733	16.2416	0.1382	14.6872	0.2756
A25	2.1157	0.0195	0.4329	0.0216	0.0990	0.0128	1.1937	0.0404	1.4520	0.0924
A26	3.8795	0.0262	2.8284	0.0547	4.6628	0.0862	3.2612	0.0662	2.4292	0.1191
A34	0.1508	0.0052	3.5228	0.0610	1.3529	0.0470	0.4928	0.0260	0.3150	0.0432
A43	0.0018	0.0006	0.0334	0.0060	0.0231	0.0062	0.0055	0.0028	0.0059	0.0059
A66	0.0173	0.0018	0.2233	0.0155	0.0478	0.0089	0.0399	0.0074	0.0534	0.0178
A10 unsplit	0.0790	0.0038	0.0939	0.0101	0.1255	0.0144	0.0647	0.0094	0.0298	0.0133
A10 total	6.2441	0.0328	7.1348	0.0850	6.3111	0.0993	5.0578	0.0816	4.2853	0.1565
A29	3.5796	0.0252	3.2071	0.0582	1.1233	0.0429	4.5156	0.0774	3.4345	0.1410
A30	2.5067	0.0212	13.0969	0.1129	2.2025	0.0598	4.4873	0.0772	2.5314	0.1215
A31	2.7386	0.0221	1.6556	0.0420	3.6005	0.0761	4.8328	0.0800	6.0881	0.1855
A32	3.6956	0.0256	1.5384	0.0405	1.0331	0.0411	2.7064	0.0604	2.5521	0.1220
A33	1.2080	0.0148	6.5607	0.0822	9.2701	0.1191	2.6593	0.0599	1.0754	0.0796
A74	0.0277	0.0022	1.9949	0.0461	0.0561	0.0096	0.2027	0.0167	0.1068	0.0252
A19 unsplit	0.0567	0.0032	0.2057	0.0149	0.0990	0.0128	0.1211	0.0129	0.0475	0.0168
A19 total	13.8129	0.0468	28.2593	0.1504	17.3846	0.1555	19.5252	0.1481	15.8358	0.2832
AX	0.8204	0.0297	4.9506	0.0963	2.9916	0.1177	1.6332	0.0878	1.8454	0.1925
a Gene frequency.
b Standard error.

[0086]

TABLE 9
Estimated gene frequencies for HLA-B antigens
	CAU	AFR	ASI	LAT	NAT
Antigen	Gfa	SEb	Gf	SE	Gf	SE	Gf	SE	Gf	SE
B7	12.1782	0.0445	10.5960	0.1024	4.2691	0.0827	6.4477	0.0918	10.9845	0.2432
B8	9.4077	0.0397	3.8315	0.0634	1.3322	0.0467	3.8225	0.0715	8.5789	0.2176
B13	2.3061	0.0203	0.8103	0.0295	4.9222	0.0886	1.2699	0.0416	1.7495	0.1013
B14	4.3481	0.0277	3.0331	0.0566	0.5004	0.0287	5.4166	0.0846	2.9823	0.1316
B18	4.7980	0.0290	3.2057	0.0582	1.1246	0.0429	4.2349	0.0752	3.3422	0.1391
B27	4.3831	0.0278	1.2918	0.0372	2.2355	0.0603	2.3724	0.0567	5.1970	0.1721
B35	9.6614	0.0402	8.5172	0.0927	8.1203	0.1122	14.6516	0.1329	10.1198	0.2345
B37	1.4032	0.0159	0.5916	0.0252	1.2327	0.0449	0.7807	0.0327	0.9755	0.0759
B41	0.9211	0.0129	0.8183	0.0296	0.1303	0.0147	1.2818	0.0418	0.4766	0.0531
B42	0.0608	0.0033	5.6991	0.0768	0.0841	0.0118	0.5866	0.0284	0.2856	0.0411
B46	0.0099	0.0013	0.0151	0.0040	4.9292	0.0886	0.0234	0.0057	0.0238	0.0119
B47	0.2069	0.0061	0.1305	0.0119	0.0956	0.0126	0.1832	0.0159	0.2139	0.0356
B48	0.0865	0.0040	0.1316	0.0119	2.0276	0.0575	1.5915	0.0466	1.0267	0.0778
B53	0.4620	0.0092	10.9529	0.1039	0.4315	0.0266	1.6982	0.0481	1.0804	0.0798
B59	0.0020	0.0006	0.0032	0.0019	0.4277	0.0265	0.0055	0.0028	0c	—
B67	0.0040	0.0009	0.0086	0.0030	0.2276	0.0194	0.0055	0.0028	0.0059	0.0059
B70	0.3270	0.0077	7.3571	0.0866	0.8901	0.0382	1.9266	0.0512	0.6901	0.0639
B73	0.0108	0.0014	0.0032	0.0019	0.0132	0.0047	0.0261	0.0060	0c	—
B51	5.4215	0.0307	2.5980	0.0525	7.4751	0.1080	6.8147	0.0943	6.9077	0.1968
B52	0.9658	0.0132	1.3712	0.0383	3.5121	0.0752	2.2447	0.0552	0.6960	0.0641
B5 unsplit	0.1565	0.0053	0.1522	0.0128	0.1288	0.0146	0.1546	0.0146	0.1307	0.0278
B5 total	6.5438	0.0435	4.1214	0.0747	11.1160	0.1504	9.2141	0.1324	7.7344	0.2784
B44	13.4838	0.0465	7.0137	0.0847	5.6807	0.0948	9.9253	0.1121	11.8024	0.2511
B45	0.5771	0.0102	4.8069	0.0708	0.1816	0.0173	1.8812	0.0506	0.7603	0.0670
B12 unsplit	0.0788	0.0038	0.0280	0.0055	0.0049	0.0029	0.0193	0.0051	0.0654	0.0197
B12 total	14.1440	0.0474	11.8486	0.1072	5.8673	0.0963	11.8258	0.1210	12.6281	0.2584
B62	5.9117	0.0320	1.5267	0.0404	9.2249	0.1190	4.1825	0.0747	6.9421	0.1973
									0.3738
B63	0.4302	0.0088	1.8865	0.0448	0.4438	0.0270	0.8083	0.0333	0.0356	0.0471
B75	0.0104	0.0014	0.0226	0.0049	1.9673	0.0566	0.1101	0.0123	0	0.0145
B76	0.0026	0.0007	0.0065	0.0026	0.0874	0.0120	0.0055	0.0028	0c	—
B77	0.0057	0.0010	0.0119	0.0036	0.0577	0.0098	0.0083	0.0034	0.0059	0.0059
B15 unsplit	0.1305	0.0049	0.0691	0.0086	0.4301	0.0266	0.1820	0.0158	0.0715	0.0206
B15 total	6.4910	0.0334	3.5232	0.0608	12.2112	0.1344	5.2967	0.0835	7.4290	0.2035
B38	2.4413	0.0209	0.3323	0.0189	3.2818	0.0728	1.9652	0.0517	1.1017	0.0806
B39	1.9614	0.0188	1.2893	0.0371	2.0352	0.0576	6.3040	0.0909	4.5527	0.1615
B16 unsplit	0.0638	0.0034	0.0237	0.0051	0.0644	0.0103	0.1226	0.0130	0.0593	0.0188
B16 total	4.4667	0.0280	1.6453	0.0419	5.3814	0.0921	8.3917	0.1036	5.7137	0.1797
B57	3.5955	0.0252	5.6746	0.0766	2.5782	0.0647	2.1800	0.0544	2.7265	0.1260
B58	0.7152	0.0114	5.9546	0.0784	4.0189	0.0803	1.2481	0.0413	0.9398	0.0745
B17 unsplit	0.2845	0.0072	0.3248	0.0187	0.3751	0.0248	0.1446	0.0141	0.2674	0.0398
B17 total	4.5952	0.0284	11.9540	0.1076	6.9722	0.1041	3.5727	0.0691	3.9338	0.1503
B49	1.6452	0.0172	2.6286	0.0528	0.2440	0.0200	2.3353	0.0562	1.5462	0.0953
B50	1.0580	0.0138	0.8636	0.0304	0.4421	0.0270	1.8883	0.0507	0.7862	0.0681
B21 unsplit	0.0702	0.0036	0.0270	0.0054	0.0132	0.0047	0.0771	0.0103	0.0356	0.0145
B21 total	2.7733	0.0222	3.5192	0.0608	0.6993	0.0339	4.3007	0.0755	2.3680	0.1174
B54	0.0124	0.0015	0.0183	0.0044	2.6873	0.0660	0.0289	0.0063	0.0534	0.0178
B55	1.9046	0.0185	0.4895	0.0229	2.2444	0.0604	0.9515	0.0361	1.4054	0.0909
B56	0.5527	0.0100	0.2686	0.0170	0.8260	0.0368	0.3596	0.0222	0.3387	0.0448
B22 unsplit	0.1682	0.0055	0.0496	0.0073	0.2730	0.0212	0.0372	0.0071	0.1246	0.0272
B22 total	2.0852	0.0217	0.8261	0.0297	6.0307	0.0971	1.3771	0.0433	1.9221	0.1060
B60	5.2222	0.0302	1.5299	0.0404	8.3254	0.1135	2.2538	0.0553	5.7218	0.1801
B61	1.1916	0.0147	0.4709	0.0225	6.2072	0.0989	4.6691	0.0788	2.6023	0.1231
B40 unsplit	0.2696	0.0070	0.0388	0.0065	0.3205	0.0230	0.2473	0.0184	0.2271	0.0367
B40 total	6.6834	0.0338	2.0396	0.0465	14.8531	0.1462	7.1702	0.0963	8.5512	0.2168
BX	1.0922	0.0252	3.5258	0.0802	3.8749	0.0988	2.5266	0.0807	1.9867	0.1634
a Gene frequency.
b Standard error.
c The observed gene count was zero.

[0087]

TABLE 10
Estimated gene frequencies of HLA-DR antigens
	CAU	AFR	ASI	LAT	NAT
Antigen	Gfa	SEb	Gf	SE	Gf	SE	Gf	SE	Gf	SE
DR1	10.2279	0.0413	6.8200	0.0832	3.4628	0.0747	7.9859	0.1013	8.2512	0.2139
DR2	15.2408	0.0491	16.2373	0.1222	18.6162	0.1608	11.2389	0.1182	15.3932	0.2818
DR3	10.8708	0.0424	13.3080	0.1124	4.7223	0.0867	7.8998	0.1008	10.2549	0.2361
DR4	16.7589	0.0511	5.7084	0.0765	15.4623	0.1490	20.5373	0.1520	19.8264	0.3123
DR6	14.3937	0.0479	18.6117	0.1291	13.4471	0.1404	17.0265	0.1411	14.8021	0.2772
DR7	13.2807	0.0463	10.1317	0.0997	6.9270	0.1040	10.6726	0.1155	10.4219	0.2378
DR8	2.8820	0.0227	6.2673	0.0800	6.5413	0.1013	9.7731	0.1110	6.0059	0.1844
DR9	1.0616	0.0139	2.9646	0.0559	9.7527	0.1218	1.0712	0.0383	2.8662	0.1291
DR10	1.4790	0.0163	2.0397	0.0465	2.2304	0.0602	1.8044	0.0495	1.0896	0.0801
DR11	9.3180	0.0396	10.6151	0.1018	4.7375	0.0869	7.0411	0.0955	5.3152	0.1740
DR12	1.9070	0.0185	4.1152	0.0655	10.1365	0.1239	1.7244	0.0484	2.0132	0.1086
DR5 unsplit	1.2199	0.0149	2.2957	0.0493	1.4118	0.0480	1.8225	0.0498	1.6769	0.0992
DR5 total	12.4449	0.0045	17.0260	0.1243	16.2858	0.1516	10.5880	0.1148	9.0052	0.2218
DRX	1.3598	0.0342	0.8853	0.0760	2.5521	0.1089	1.4023	0.0930	2.0834	0.2037
a Gene frequency.
b Standard error.

[0088] It can be desirable to express housekeeping peptides in the context of a larger protein. Processing can be detected even when a small number of amino acids are present beyond the terminus of an epitope. Small peptide hormones are usually proteolytically processed from longer translation products, often in the size range of approximately 60-120 amino acids. This fact has led some to assume that this is the minimum size that can be efficiently translated. In some embodiments, the housekeeping peptide can be embedded in a translation product of at least about 60 amino acids, in others 70, 80, 90 amino acids, and in still others 100, 110 or 120 amino acids, for example. In other embodiments the housekeeping peptide can be embedded in a translation product of at least about 50, 30, or 15 amino acids.

[0089] Due to differential proteasomal processing, the immunoproteasome of the pAPC produces peptides that are different from those produced by the housekeeping proteasome in peripheral body cells. Thus, in expressing a housekeeping peptide in the context of a larger protein, it is preferably expressed in the pAPC in a context other than its full-length native sequence, because, as a housekeeping epitope, it is generally only efficiently processed from the native protein by the housekeeping proteasome, which is not active in the pAPC. In order to encode the housekeeping epitope in a DNA sequence encoding a larger polypeptide, it is useful to find flanking areas on either side of the sequence encoding the epitope that permit appropriate cleavage by the immunoproteasome in order to liberate that housekeeping epitope. Such a sequence promoting appropriate processing is referred to hereinafter as having substrate or liberation sequence function. Altering flanking amino acid residues at the N-terminus and C-terminus of the desired housekeeping epitope can facilitate appropriate cleavage and generation of the housekeeping epitope in the pAPC. Sequences embedding housekeeping epitopes can be designed de novo and screened to determine which can be successfully processed by immunoproteasomes to liberate housekeeping epitopes.

[0090] Alternatively, another strategy is very effective for identifying sequences allowing production of housekeeping epitopes in APC. A contiguous sequence of amino acids can be generated from head to tail arrangement of one or more housekeeping epitopes. A construct expressing this sequence is used to immunize an animal, and the resulting T cell response is evaluated to determine its specificity to one or more of the epitopes in the array. These immune responses indicate housekeeping epitopes that are processed in the pAPC effectively. The necessary flanking areas around this epitope are thereby defined. The use of flanking regions of about 4-6 amino acids on either side of the desired peptide can provide the necessary information to facilitate proteasome processing of the housekeeping epitope by the immunoproteasome. Therefore, a substrate or liberation sequence of approximately 16-22 amino acids can be inserted into, or fused to, any protein sequence effectively to result in that housekeeping epitope being produced in an APC. In some embodiments, a broader context of a substrate sequence can also influence processing. In such embodiments, comparisons of a liberaton sequence in a variety of contexts can be useful in further optimizing a particular substrate sequence. In alternate embodiments the whole head-to-tail array of epitopes, or just the epitopes immediately adjacent to the correctly processed housekeeping epitope can be similarly transferred from a test construct to a vaccine vector. [0084] In a preferred embodiment, the housekeeping epitopes can be embedded between known immune epitopes, or segments of such, thereby providing an appropriate context for processing. The abutment of housekeeping and immune epitopes can generate the necessary context to enable the immunoproteasome to liberate the housekeeping epitope, or a larger fragment, preferably including a correct C-terminus. It can be useful to screen constructs to verify that the desired epitope is produced. The abutment of housekeeping epitopes can generate a site cleavable by the immunoproteasome. Some embodiments of the invention employ known epitopes to flank housekeeping epitopes in test substrates; in others, screening as described below is used, whether the flanking regions are arbitrary sequences or mutants of the natural flanking sequence, and whether or not knowledge of proteasomal cleavage preferences are used in designing the substrates.

[0091] Cleavage at the mature N-terminus of the epitope, while advantageous, is not required, since a variety of N-terminal trimming activities exist in the cell that can generate the mature N-terminus of the epitope subsequent to proteasomal processing. It is preferred that such N-terminal extension be less than about 25 amino acids in length and it is further preferred that the extension have few or no proline residues. Preferably, in screening, consideration is given not only to cleavage at the ends of the epitope (or at least at its C-terminus), but consideration also can be given to ensure limited cleavage within the epitope.

[0092] Shotgun approaches can be used in designing test substrates and can increase the efficiency of screening. In one embodiment multiple epitopes can be assembled one after the other, with individual epitopes possibly appearing more than once. The substrate can be screened to determine which epitopes can be produced. In the case where a particular epitope is of concern, a substrate can be designed in which it appears in multiple different contexts. When a single epitope appearing in more than one context is liberated from the substrate additional secondary test substrates, in which individual instances of the epitope are removed, disabled, or are unique, can be used to determine which are being liberated and truly confer substrate or liberation sequence function.

[0093] Several readily practicable screens exist. A preferred in vitro screen utilizes proteasomal digestion analysis, using purified immunoproteasomes, to determine if the desired housekeeping epitope can be liberated from a synthetic peptide embodying the sequence in question. The position of the cleavages obtained can be determined by techniques such as mass spectrometry, HPLC, and N-terminal pool sequencing; as described in greater detail in U.S. patent application Ser. Nos. 09/561,074, 09/560,465 and 10/117,937, and Provisional U.S. Patent Application Nos. 60/282,211, 60/337,017, and 60/363, 210, which were all cited and incorporated by reference above.

[0094] Alternatively, in vivo and cell-based screens such as immunization or target sensitization can be employed. For immunization a nucleic acid construct capable of expressing the sequence in question is used. Harvested CTL can be tested for their ability to recognize target cells presenting the housekeeping epitope in question. Such targets cells are most readily obtained by pulsing cells expressing the appropriate MHC molecule with synthetic peptide embodying the mature housekeeping epitope. Alternatively, immunization can be carried out using cells known to express housekeeping proteasome and the antigen from which the housekeeping epitope is derived, either endogenously or through genetic engineering. To use target sensitization as a screen, CTL, or preferably a CTL clone, that recognizes the housekeeping epitope can be used. In this case it is the target cell that expresses the embedded housekeeping epitope (instead of the pAPC during immunization) and it must express immunoproteasome. Generally, the cell or target cell can be transformed with an appropriate nucleic acid construct to confer expression of the embedded housekeeping epitope. Loading with a synthetic peptide embodying the embedded epitope using peptide loaded liposomes, or complexed with cationic lipid protein transfer reagents such as BIOPORTER™ (Gene Therapy Systems, San Diego, Calif.), represents an alternative.

[0095] Once sequences with substrate or liberation sequence function are identified they can be encoded in nucleic acid vectors, chemically synthesized, or produced recombinantly. In any of these forms they can be incorporated into immunogenic compositions. Such compositions can be used in vitro in vaccine development or in the generation or expansion of CTL to be used in adoptive immunotherapy. In vivo they can be used to induce, amplify or sustain and active immune response. The uptake of polypeptides for processing and presentation can be greatly enhanced by packaging with cationic lipid, the addition of a tract of cationic amino acids such as poly-L-lysine (Ryser, H. J. et al., J. Cell Physiol. 113:167-178, 1982; Shen, W. C. & Ryser, H. J., Proc. Natl. Aced. Sci. USA 75:1872-1876, 1978), the incorporation into branched structures with importation signals (Sheldon, K. et al., Proc. Natl. Aced. Sci. USA 92:2056-2060, 1995), or mixture with or fusion to polypeptides with protein transfer function including peptide carriers such as pep-1 (Morris, M. C., et al., Nat. Biotech. 19:1173-1176, 2001), the PreS2 translocation motif of hepatitis B virus surface antigen, VP22 of herpes viruses, and HIV-TAT protein (Oess, S. & Hildt, E., Gene Ther. 7:750-758, 2000; Ford, K. G., et al., Gene Ther. 8:1-4, 2001; Hung, C. F. et al., J. Virol. 76:2676-2682, 2002; Oliveira, S. C., et a;. Hum. Gene Ther. 12:1353-1359, 2001; Normand, N. et al., J. Biol. Chem. 276:15042-15050, 2001; Schwartz, J. J. & Zhang, S., Curr. Opin. Mol. Ther. 2:162-167, 2000; Elliot G., 7 Hare, P. Cell 88:223-233, 1997), among other methodologies. Particularly for fusion proteins the immunogen can be produced in culture and the purified protein administered or, in the alternative, the nucleic acid vector can be administered so that the immunogen is produced and secreted by cells transformed in vivo. In either scenario the transport function of the fusion protein facilitates uptake by pAPC.

EXAMPLES

Example 1

[0096] A recombinant DNA plasmid vaccine, pMA2M, which encodes one polypeptide with an HLA A2-specific CTL epitope ELAGIGILTV (SEQ ID NO. 1) from melan-A (26-35A27L), and a portion (amino acids 31-96) of melan-A (SEQ ID NO. 2) including the epitope clusters at amino acids 31-48 and 56-69, was constructed. These clusters were previously disclosed in U.S. patent application Ser. No. 09/561,571 entitled EPITOPE CLUSTERS incorporated by reference above. Flanking the defined melan-A CTL epitope are short amino acid sequences derived from human tyrosinase (SEQ ID NO. 3) to facilitate liberation of the melan-A housekeeping epitope by processing by the immunoproteasome. In addition, these amino acid sequences represent potential CTL epitopes themselves. The cDNA sequence for the polypeptide in the plasmid is under the control of promoter/enhancer sequence from cytomegalovirus (CMVp) (see FIG. 1), which allows efficient transcription of messenger for the polypeptide upon uptake by APCs. The bovine growth hormone polyadenylation signal (BGH polyA) at the 3′ end of the encoding sequence provides a signal for polyadenylation of the messenger to increase its stability as well as for translocation out of nucleus into the cytoplasm for translation. To facilitate plasmid transport into the nucleus after uptake, a nuclear import sequence (NIS) from simian virus 40 (SV40) has been inserted in the plasmid backbone. The plasmid carries two copies of a CpG immunostimulatory motif, one in the NIS sequence and one in the plasmid backbone. Lastly, two prokaryotic genetic elements in the plasmid are responsible for amplification in E. coli, the kanamycin resistance gene (Kan R) and the pMB1 bacterial origin of replication.

[0097] SUBSTRATE or LIBERATION Sequence

[0098] The amino acid sequence of the encoded polypeptide (94 amino acid residues in length) (SEQ ID NO. 4) containing a 28 amino acid substrate or liberation sequence at its N-terminus (SEQ ID NO. 5) is given below:

MLLAVLYCL-ELAGIGTLTV-YMDGTMSQV-
GILTVILGVLLLIGCWYCRRRNGYRALMDKSLHVGTQCALTRRCPQEGFDHRDSKVSLQ
EKNCEPV

[0099] The first 9 amino acid residues are derived from tyrosinasel1-9 (SEQ ID NO. 6), the next ten constitute melan-A (26-35A27L) (SEQ ID NO. 1), and amino acid residues 20 to 29 are derived from tyrosinase369-377 (SEQ ID NO. 7). These two tyrosinase nonamer sequences both represent potential HLA A2-specific CTL epitopes. Amino acid residues 10-19 constitute melan-A (26-35A27L) an analog of an HLA A2-specific CTL epitope from melan-A, EAAGIGILTV (SEQ ID NO. 8), with an elevated potency in inducing CTL responses during in vitro immunization of human PBMC and in vivo immunization in mice. The segment of melan-A constituting the rest of the polypeptide (amino acid residues 30 to 94) contain a number of predicted HLA A2-specific epitopes, including the epitope clusters cited above, and thus can be useful in generating a response to immune epitopes as described at length in the patent applications ‘Epitope Synchronization in Antigen Presenting Cells’ and ‘Epitope Clusters’ cited and incorporated by reference above. This region was also included to overcome any difficulties that can be associated with the expression of shorter sequences. A drawing of pMA2M is shown in FIG. 1.

[0100] Plasmid Construction

[0101] A pair of long complementary oligonucleotides was synthesized which encoded the first 30 amino acid residues. In addition, upon annealing, these oligonucleotides generated the cohensive ends of Afl II at the 5′ end and that of EcoR I at the 3′ end. The melan A31-96 region was amplified with PCR using oligonucleotides carrying restriction sites for EcoR I at the 5′ end and Not I at the 3′ end. The PCR product was digested with EcoR I and Not I and ligated into the vector backbone, described in Example 1, that had been digested with Afl II and Not I, along with the annealed oligonucleotides encoding the amino terminal region in a three-fragment ligation. The entire coding sequence was verified by DNA sequencing. The sequence of the entire insert, from the Afl II site at the 5′ end to the Not I site at the 3′ end is disclosed as SEQ ID NO. 9. Nucleotides 12-293 encode the polypeptide.

Example 2

[0102] Three vectors containing melan-A (26-35A27L) (SEQ ID NO. 1) as an embedded housekeeping epitope were tested for their ability to induce a CTL response to this epitope in HLA-A2 transgenic HHD mice (Pascolo et al. J Exp. Med. 185:2043-2051, 1997). One of the vectors was pMA2M described above (called pVAXM3 in FIG. 3). In pVAXM2 the same basic group of 3 epitopes was repeated several times with the flanking epitopes truncated by differing degrees in the various repeats of the array. Specifically the cassette consisted of:

M-Tyr(5-9)-ELA-Tyr(369-373)- (SEQ ID NO. 10)
Tyr(4-9)-ELA-Tyr(369-374)-Tyr
(3-9)-ELA-Tyr(369-375)-Tyr(2-
9)-ELA

[0103] where ELA represents melan-A (26-35A27L) (SEQ ID NO. 1). This cassette was inserted in the same plasmid backbone as used for pVAXM3. The third, pVAXM1 is identical to pVAXM2 except that the epitope array is followed by an IRES (internal ribosome entry site for encephalomyocarditis virus) linked to a reading frame encoding melan-A 31-70.

[0104] Four groups of three HHD A2.1 mice were injected intranodally in surgically exposed inguinal lymph nodes with 25 μl of 1 mg/ml plasmid DNA in PBS on days 0, 3, and 6, each group receiving one of the three vectors or PBS alone. On day 14 the spleens were harvested and restimulated in vitro one time with 3-day LPS blasts pulsed with peptide (melan-A (26-35A27L)(SEQ ID NO. 1)). The in vitro cultures were supplemented with Rat T-Stim (Collaborative Biomedical Products) on the 3rd day and assayed for cytolytic activity on the 7th day using a standard 51Cr-release assay. FIGS. 2 to 5 show % specific lysis obtained using the cells immunized with PBS, pVAXM1, pVAXM2, and pVAXM3, respectively on T2 target cells and T2 target cells pulsed with melan-A (26-35A27L) (ELA) (SEQ ID NO. 1). All three vectors generated strong CTL responses. These data indicated that the plasmids have been taken up by APCs, the encoded polypeptide has been synthesized and proteolytically processed to produce the decamer epitope in question (that is, it had substrate or liberation sequence function), and that the epitope became HLA-A2 bound for presentation. Also, an isolated variant of pVAXM2, that terminates after the 55th amino acid, worked similarly well as the full length version (data not shown). Whether other potential epitopes within the expression cassette can also be produced and be active in inducing CTL responses can be determined by testing for CTL activity against target cells pulsed with corresponding synthetic peptides.

Example 3

[0105] An NY-ESO-1 (SEQ ID NO. 11) SUBSTRATE/LIBERATION Sequence

[0106] Six other epitope arrays were tested leading to the identification of a substrate/liberation sequence for the housekeeping epitope NY-ESO-1157-165 (SEQ ID NO. 12). The component epitopes of the arrays were:

SSX-241-49: KASEKTFYV	Array element A	(SEQ ID NO. 13)
NY-ESO-1157-165: SLLMWITQC	Array element B	(SEQ ID NO. 12)
NY-ESO-1163-171: TQCFLPVFL	Array element C	(SEQ ID NO. 14)
PSMA288-297: GLPSIPVHPI	Array element D	(SEQ ID NO. 15)
TYR4-9: AVLYCL	Array element B	(SEQ ID NO. 16)

[0107] The six arrays had the following arrangements of elements after starting with an initiator methionine:

pVAX-PC-A: B-A-D-D-A-B-A-A
pVAX-PC-B: D-A-B-A-A-D-B-A
pVAX-PC-C: E-A-D-B-A-B-E-A-A
pVAX-BC-A: B-A-C-B-A-A-C-A
pVAX-BC-B: C-A-B-C-A-A-B-A
pVAX-BC-C: E-A-A-B-C-B-A-A

[0108] These arrays were inserted into the same vector backbone described in the examples above. The plasmid vectors were used to immunize mice essentially as described in Example 2 and the resulting CTL were tested for their ability to specifically lyse target cells pulsed with the peptide NY-ESO-1 157-165, corresponding to element B above. Both pVAX-PC-A and pVAX-BC-A were found to induce specific lytic activity. Comparing the contexts of the epitope (element B) in the various arrays, and particularly between pVAX-PC-A and pVAX-BC-A, between pVAX-PC-A and pVAX-PC-B, and between pVAX-BC-A and pVAX-BC-C, it was concluded that it was the first occurrence of the epitope in pVAX-PC-A and pVAX-BC-A that was being correctly processed and presented. In other words an initiator methionine followed by elements B-A constitute a substrate/liberation sequence for the presentation of element B. On this basis a new expression cassette for use as a vaccine was constructed encoding the following elements:

[0109] An initiator methionine,

[0110] NY-ESO-1157-165 (bold)—a housekeeping epitope,

[0111] SSX241-49 (italic)—providing appropriate context for processing, and

[0112] NY-ESO-177-180—to avoid “short sequence” problems and provide immune epitopes.

[0113] Thus the construct encodes the amino acid sequence:

[0114] M-SLLMWITQC-KASEKIFYV-RCGARGPESRLLEFYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEFTVSGNILTIRL TAADHRQLQLSISSCLQQLSLLMWITQCFLPVFLAQPPSGQRR (SEQ ID NO. 17) and MSLLMWITQCKASEKIFYV (SEQ ID NO. 18) constitutes the liberation or substrate sequence. A polynucleotide encoding SEQ ID NO. 17 (SEQ ID NO. 19: nucleotides 12-380) was inserted into the same plasmid backbone as used for pMA2M generating the plasmid pN157.

Example 4

[0115] A construct similar to pN157 containing the whole epitope array from pVAX-PC-A was also made and designated pBPL. Thus the encoded amino acid sequence in pBPL is:

M-SLLMWITQC-KASEKIFYV-GLPSIPVHPI-GLPSIPVHPI-KASEKIFYV-SLLMWITQC-KASEKIFYV-KASEKIFYV-RCGARGPESRLLEFYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEFTVSGNILTTRL (SEQ ID NO. 20)
TAADHRQLQLSISSCLQQLSLLMWITQCFLPVFLAQPPSGQRR.

[0116] SEQ ID NO. 21 is the polynucleotide encoding SEQ ID NO. 20 used in pBPL.

[0117] A portion of SEQ ID NO. 20, IKASEKIFYVSLLMWITQCKASEKIFYVK (SEQ ID NO. 22) was made as a synthetic peptide and subjected to in vitro proteasomal digestion analysis with human immunoproteasome, utilizing both mass spectrometry and N-terminal pool sequencing. The identification of a cleavage after the C residue indicates that this segment of the construct can function as a substrate or liberation sequence for NY-ESO-1157-165 (SEQ ID NO. 12) epitope (see FIG. 6). FIG. 7 shows the differential processing of the SLLMWITQC epitope (SEQ ID NO. 12) in its native context where the cleavage following the C is more efficiently produced by housekeeping than immunoproteasome. The immunoproteasome also produces a major cleavage internal to the epitope, between the T and the Q when the epitope is in its native context, but not in the context of SEQ ID NO. 22 (compare FIG. 6 and 7).

Example 5

[0118] Screening of further epitope arrays led to the identification of constructs promoting the expression of the epitope SSX-241-49 (SEQ ID NO. 13). In addition to some of the array elements defined in Example 3, the following additional elements were also used:

	SSX-457-65:
	VMTKLGFKV	Array element F.	(SEQ ID NO. 23)
	PSMA730-739:
	RQIYVAAFTV	Array element G.	(SEQ ID NO. 24)

[0119] A construct, denoted CTLA02, encoding an initiator methionine and the array F-A-G-D-C-F-G-A, was found to successfully immunize HLA-A2 transgenic mice to generate a CTL response recognizing the peptide SSX-241-49 (SEQ ID NO. 13).

[0120] As described above, it can be desirable to combine a sequence with substrate or liberation sequence function with one that can be processed into immune epitopes. Thus SSX-215-183 (SEQ ID NO. 25) was combined with all or part of the array as follows:

CTLS1:
F-A-G-D-C-F-G-A-SSX-215-183 (SEQ ID NO. 26)
CTLS2:
SSX-215-183-F-A-G-D-C-F-G-A (SEQ ID NO. 27)
CTLS3:
F-A-G-D-SSX-215-183         (SEQ ID NO. 28)
CTLS4:
SSX-215-183-C-F-G-A         (SEQ ID NO. 29).

[0121] All of the constructs except CTLS3 were able to induce CTL recognizing the peptide SSX-241-49 (SEQ ID NO. 13). CTLS3 was the only one of these four constructs which did not include the second element A from CTLA02 suggesting that it was this second occurrence of the element that provided substrate or liberation sequence function. In CTLS2 and CTLS4 the A element is at the C-terminal end of the array, as in CTLA02. In CTLS1 the A element is immediately followed by the SSX-215-183 segment which begins with an alanine, a residue often found after proteasomal cleavage sites (Toes, R. E. M., et al., J. Exp. Med. 194:1-12, 2001). SEQ ID NO. 30 is the polynucleotide sequence encoding SEQ ID NO. 26 used in CTLS1, also called pCBP.

[0122] A portion of CTLS1 (SEQ ID NO. 26), encompassing array elements F-A-SSX-215-23 with the sequence RQIYVAAFTV-KASEKIFYV-AQIPEKIQK (SEQ ID NO. 31), was made as a synthetic peptide and subjected to in vitro proteasomal digestion analysis with human immunoproteasome, utilizing both mass spectrometry and N-terminal pool sequencing. The observation that the C-terminus of the SSX-241-49 epitope (SEQ ID NO. 13) was generated (see FIG. 8) provided further evidence in support of substrate or liberation sequence function. The data in FIG. 9 showed the differential processing of the SSX-241-49 epitope, KASEKIFYV (SEQ ID NO. 13), in its native context, where the cleavage following the V was the predominant cleavage produced by housekeeping proteasome, while the immunoproteasome had several major cleavage sites elsewhere in the sequence. By moving this epitope into the context provided by SEQ ID NO. 31 the desired cleavage became a major one and its relative frequency compared to other immunoproteasome cleavages was increased (compare FIGS. 8 and 9). The data in FIG. 8B also showed the similarity in specificity of mouse and human immunoproteasome lending support to the usefulness of the transgenic mouse model to predict human antigen processing.

Example 6

[0123] Screening also revealed substrate or liberation sequence function for a tyrosinase epitope, Tyr207-215 (SEQ ID NO. 32), as part of an array consisting of the sequence [Tyr1-17-Tyr207-215]4, [MLLAVLYCLLWSFQTSA-FLPWHRLFL]4, (SEQ ID NO. 33). The same vector backbone described above was used to express this array. This array differs from those of the other examples in that the Tyr1-17 segment, which was included as a source of immune epitopes, is used as a repeated element of the array. This is in contrast with the pattern shown in the other examples where sequence included as a source of immune epitopes and/or length occurred a single time at the beginning or end of the array, the remainder of which was made up of individual epitopes or shorter sequences.

[0124] Plasmid Construction

[0125] The polynucleotide encoding SEQ ID NO. 33 was generated by assembly of annealed synthetic oligonucleotides. Four pairs of complementary oligonucleotides were synthesized which span the entire coding sequence with cohesive ends of the restriction sites of Afl II and EcoR I at either terminus. Each complementary pair of oligonucleotides were first annealed, the resultant DNA fragments were ligated stepwise, and the assembled DNA fragment was inserted into the same vector backbone described above pre-digested with Afl II/EcoR I. The construct was called CTLT2/pMEL and SEQ ID NO. 34 is the polynucleotide sequence used to encode SEQ ID NO. 33.

Example 7

[0126] Administration of a DNA Plasmid Formulation of a Immunotherapeutic for Melanoma to Humans

[0127] An MA2M melanoma vaccine with a sequence as described in Example 1 above, was formulated in 1% Benzyl alcohol, 1% ethyl alcohol, 0.5mM EDTA, citrate-phosphate, pH 7.6. Aliquots of 200, 400, and 600 μg DNA/ml were prepared for loading into MINIMED 407C infusion pumps. The catheter of a SILHOUETTE infusion set was placed into an inguinal lymph node visualized by ultrasound imaging. The pump and infusion set assembly was originally designed for the delivery of insulin to diabetics. The usual 17mm catheter was substituted with a 31 mm catheter for this application. The infusion set was kept patent for 4 days (approximately 96 hours) with an infusion rate of about 25 μl/hour resulting in a total infused volume of approximately 2.4 ml. Thus the total administered dose per infusion was approximately 500, and 1000 μg; and can be 1500 μg, respectively, for the three concentrations described above. Following an infusion, subjects were given a 10 day rest period before starting a subsequent infusion. Given the continued residency of plasmid DNA in the lymph node after administration and the usual kinetics of CTL response following disappearance of antigen, this schedule will be sufficient to maintain the immunologic CTL response.

Example 8

[0128] SEQ ID NO. 22 is made as a synthetic peptide and packaged with a cationic lipid protein transfer reagent. The composition is infused directly into the inguinal lymph node (see example 7) at a rate of 200 to 600 μg of peptide per day for seven days, followed by seven days rest. An initial treatment of 3-8 cycles are conducted.

Example 9

[0129] A fusion protein is made by adding SEQ ID NO. 34 to the 3′ end of a nucleotide sequence encoding herpes simplex virus 1 VP22 (SEQ ID NO. 42) in an appropriate mammalian expression vector; the vector used above is suitable. The vector is used to transform HEK 293 cells and 48 to 72 hours later the cells are pelleted, lysed and a soluble extract prepared. The fusion protein is purified by affinity chromatagraphy using an anti-VP22 monoclonal antibody. The purified fusion protein is administered intranodally at a rate of 10 to 100 μg per day for seven days, followed by seven days rest. An initial treatment of 3-8 cycles are conducted.

[0130] All references mentioned herein are hereby incorporated by reference in their entirety. Further, the present invention can utilize various aspects of the following, which are all incorporated by reference in their entirety: U.S. patent application Ser. Nos. 09/380,534, filed on Sep. 1, 1999, entitled A METHOD OF INDUCING A CTL RESPONSE; 09/776,232, filed on Feb. 2, 2001, entitled METHOD OF INDUCING A CTL RESPONSE; 09/715,835, filed on Nov. 16, 2000, entitled AVOIDANCE OF UNDESIRABLE REPLICATION INTERMEDIATES IN PLASMID PROPOGATION; 09/999,186, filed on Nov. 7, 2001, entitled METHODS OF COMMERCIALIZING AN ANTIGEN; and Provisional U.S. Patent Application No. 60/274,063, filed on Mar. 7, 2001, entitled ANTI-NEOVASCULAR VACCINES FOR CANCER.

TABLE 11
Partial listing of SEQ ID NOS.
1	ELAGIGILTV	melan-A 26-3 5 (A27L)
2	Melan-A protein	Accession number: NP 005502
3	Tyrosinase protein	Accession number: P14679
4	MLLAVLYCLELAGIGILTVYMDGTMSQVG	pMA2M expression product
	ILTVILGVLLLIGCWYCRRRNGYRALMDK
	SLHVGTQCALTRRCPQEGFDHRDSKVSLQ
	EKNCEPV
5	MLLAVLYCLELAGIGILTVYMDGTMSQV	Liberation or substrate sequence for SEQ ID NO. 1
		from pMA2M
6	MLLAVLYCL	tyrosinase 1-9
7	YMDGTMSQV	tyrosinase 369-377
8	EAAGIGILTV	melan-A 26-35
9	cttaagccaccatgttactagctgttttgtactgcctggaact	pMA2M insert
	agcagggatcggcatattgacagtgtatatgga
	tggaacaatgtcccaggtaggaattctgacagtgatcctggga
	gtcttactgctcatcggctgttggtattgtaga
	agacgaaatggatacagagccttgatggataaaagtcttcatg
	ttggcactcaatgtgccttaacaagaagatgcc
	cacaagaagggtttgatcatcgggacagcaaagtgtctcttca
	agagaaaaactgtgaacctgtgtagtgagcggc
	cgc
10	MVLYCLELAGIGILTVYMDGTAVLYCLEL	Epitope array from pVAXM2 and
	AGIGILTVYMDGTMLAVLYCLELAGIGILT	pVAXM1
	VYMDGTMSLLAVLYCLELAGIGILTV
11	NY-ESO-1 protein	Accession number: P78358
12	SLLMWITQC	NY-ESO-1 157-165
13	KASEKIFYV	SSX-2 41-49
14	TQCFLPVFL	NY-ESO-1 163-171
15	GLPSIPVHPI	PSMA 288-297
16	AVLYCL	tyrosinase 4-9
17	MSLLMWITQCKASEKIFYVRCGARGPESR	pN157 expression product
	LLEFYLAMPFATPMEAELARRSLAQDAPP
	LPVPGVLLKEFTVSGNTLTTRLTAADHRQL
	QLSISSCLQQLSLLMWITQCFLPVFLAQPPS
	GQRR
18	MSLLMWITQCKASEKTFYV	liberation or substrate sequence for SEQ NO. 12
		from pN157
19	cttaagccaccatgtccctgttgatgtggatcacgcagtgcaa	Insert for pN157
	agcttcggagaaaatcttctacgtacggtgcgg
	tgccagggggccggagagccgcctgcttgagttctacctcgcc
	atgcctttcgcgacacccatggaagcagagctg
	gcccgcaggagcctggcccaggatgccccaccgcttcccgtgc
	cayggytgcttctgaaggagttcactgtgtccg
	gcaacatactgactatccgactgactgctgcagaccaccgcca
	actgcayctctccatcagctcctgtctccagca
	gctttccctgttgatgtggatcacgcagtgctttctycccgtg
	tttttggctcagcctccctcagggcagaggcgc
	tagtgagaattc
20	MSLLMWITQCKASEKIFYVGLPSIPVHPIGL	pBPL expression product
	PSIPVHPIKASEKTFYVSLLMWITQCKASEK
	IFYVKASEKIFYVRCGARGPESRLLEFYLA
	MPFATPMEAELARRSLAQDAPPLPVPGVL
	LKEFTVSGNILTIRLTAADHRQLQLSISSCL
	QQLSLLMWITQCFLPVFLAQPPSGQRR
21	atgtccctgttgatgtggatcacgcaqtgcaaagcttcggaga	pBPL insert coding region
	aaatcttctatgtgggtcttccaagtattcctg
	ttcatccaattggtcttccaagtattcctgttcatccaattaa
	agcttcggagaaaatcttctatgtgtccctgtt
	gatgtggatcacgcagtgcaaagcttcggagaaaatcttctat
	gtgaaagcttcggagaaaatcttctacgtacqg
	tgcggtgccagggggccggagagccgcctgcttgagttctacc
	tcgccatgcctttcgcgacacccatggaagcag
	agctggcccgcaggagcctggcccaggatgccccaccgcttcc
	cgtgccaggggtgcttctgaaggagttcactgt
	gtccggcaacatactgactatccgactgactgctgcagaccac
	cgccaactgcagctctccatcagctcctgtctc
	cagcagctttccctgttgatgtggatcacgcagtgctttctgc
	ccgtgtttttggctcagcctccctcagggcaga
	ggcgctagtga
22	IKASEKIFYVSLLMWITQCKASEKIIFYVK	Substrate in FIG. 6
23	VMTKLGFKV	SSX-457-65
24	RQLYVAAFTV	PSMA730-739
25	AQTPEKIQKAFDDIAKYFSKEEWEKMKAS	SSX-215-183
	EKIFYVYMKRKYEAMTKLGFKATLPPFMC
	NKRAEDFQGNDLDNDPNRGNQVERPQMT
	FGRLQGISPKIMPKKPAEEGNDSEEVPEAS
	GPQNDGKELCPPGKIPTTSEKIHERSGPKRG
	EHAWTHRLRERKQLVIYEEISDP
26	MVMTKLGFKVKASEKJIFYVRQJYVAAFTV	CTLS1/pCBP expression product
	GLPSIPVHPITQCFLPVFLVMTKLGFKVRQI
	YVAETVKASEKJFYVAQTPEKIQKAFDDI
	AKYFSKEEWEKMKASEKIFYVYMKRKYE
	AMTKLGFKATLPPFMCNKRAEDFQGNDL
	DNDPNRGNQVERPQMTFGRLQGISPKIMP
	KKPAEEGNDSEEVPEASGPQNDGKELCPP
	GKPTTSEKIHERSGPKRGEHAWTHRLRER
	KQLVTYEEISDP
27	MAQIPEKIQKAFDDIAKYFSKEEWEKMKA	CTLS2 expression product
	SEKIFYVYMKRKYEAMTKLGFKATLPPFM
	CNKRAEDFQGNDLDNDPNRGNQVERPQM
	TFGRLQGISPKIMPKKPAEEGNPSEEVPEA
	SGPQNDGKELCPPGKPTTSEKIHERSGPKR
	GEHAWTHRLRERKQLVIYEEISDPVMTKL
	GFKVKASEKIFYVRQIYVAAFTVGLPSIIW
	HPITQCFLPVFLVMTKLGFKVRQIYVAAFT
	VKASEKIFYV
28	MVMTKLGFKVKASEKIFYVRQIYVAAFTV	CTLS3 expression product
	GLPSIPVHPIAQTPEKIQKAFDDIAKYFSKEE
	WEKMKASEKIFYVYMKRKYEAMTKLGF
	KATLPPFMCNKRAEDFQGNDLDNDPNRG
	NQVERPQMTFGRLQGISPKIMPKKPAEEG
	NDSEEVPEASGPQNDGKELCPPGKPTTSE
	KIHERSGPKRGEHAWTHRLRERKQLVJYE
	EISDP
29	MAQIPEKIQKAFDDIAKYFSKEEWEKMKA	CTLS4 expression product
	SEKIFYVYMKRKYEAMTKLGFKATLPPFM
	CNKRAEDFQGNPLDNDPNRGNQVERPQM
	TFGRLQGISPKIMPKKPAEEGNDSEEVPEA
	SGPQNDGKELCPPGKPTTSEKIHERSGPKR
	GEHAWTHRLRERKQLVLYEEISDPTQCFLP
	VFLVMTKLGFKVRQIYVAAFTVKASEKIF
	YV
30	atggtcatgactaaactaggtttcaaggtcaaagcttcggaga	pcBP insert coding region
	aaatcttctatgtgagacagatttatgttgcag
	ccttcacagtgggtcttccaagtattcctgttcatccaattac
	gcagtgctttctgcccgtgtttttggtcatgac
	taaactaggtttcaaggtcagacagatttatgttgcagccttc
	acagtgaaagcttcggagaaaatcttctacgta
	gctcaaataccagagaagatccaaaaggccttcgatgatattg
	ccaaatacttctctaaggaagagtgggaaaaga
	tgaaagcctcggagaaaatcttctatgtgtatatgaagagaaa
	gtatgaggctatgactaaactaggtttcaaggc
	caccctcccacctttcatgtytaataaacgggccyaagacttc
	caggggaatgatttggataatgaccctaaccgt
	gggaatcaggttgaacgtcctcagatgactttcggcaggctcc
	agggaatctccccgaagatcatgcccaagaagc
	cagcagaggaaggaaatgattcggaygaagtgccagaagcatc
	tggcccacaaaatgatgggaaagagctgtgccc
	cccgggaaaaccaactacctctgagaagattcacgagagatct
	ggacccaaaaggggggaacatgcctygacccac
	agactgcgtgagagaaaacagctggtgatttatgaagagatca
	ycgacccttagtga
31	RQIYVAAFTVKASEKTFYVAQIPEKIQK	FIG. 8 substrate/CTLS1-2
32	FLPWHRLFL	TYR207-215
33	MLLAVLYCLLWSFQTSAFLPWHRLFLMLL	CTLT2/pMEL expression product
	AVLYCLLWSFQTSAFLPWHRLFLMLLAVL
	YCLLWSFQTSAFLPWHRLFLMLLAVLYCL
	LWSFQTSAFLPWHRLFL
34	atgctcctggctgttttgtactgcctgctgtggagtttccaga	CTLT2/pMEL insert coding region
	cctccgcttttctgccttggcatagactcttct
	tgatgctcctggctgttt tgtactgcctgctgtggagtttcca
	gacctccgcttttctgccttggcatagactctt
	cttgatgctcctggctgttttgtactgcctgctgtggagtttc
	cagacctccgcttttctgccttggcatagactc
	ttcttgatgctcctggctgttttgtactgcctgctgtggagtt
	tccagacctccgcttttctgccttggcatagac
	tcttcttgtagtga
35	MELAN-A cDNA	Accession number: NM_005511
36	Tyrosinase cDNA	Accession number: NM_000372
37	NY-ESO-1 cDNA	Accession number: U87459
38	PSMA protein	Accession number: NP_004467
39	PSMA cDNA	Accession number: NM_004476
40	SSX-2 protein	Accession number: NP_003138
41	SSX-2 cDNA	Accession number: NM_003147
42	atgacctctcgccgctccgtgaagtcgggtccgcgggaggttccg	From accession number: D10879
	cgcgatgagtacgaggatctgtactacaccccgtcttcaggtatgg	Herpes Simplex virus 1 UL49 coding
	cgagtcccgatagtccgcctgacacctcccgccgtggcgccctac sequence (VP22)
	agacacgctcgcgccagaggggcgaggtccgtttcgtccagtac
	gacgagtcggattatgccctctacgggggctcgtcatccgaagac
	gacgaacacccggaggtcccccggacgcggcgtcccgtttccgg
	ggcggttttgtccggcccggggcctgcgcgggcgcctccgccac
	ccgctgggtccggaggggccggacgcacacccaccaccgcccc
	ccgggccccccgaacccagcgggtggcgactaaggcccccgcg
	gccccggcggcggagaccacccgcggcaggaaatcggcccag
	ccagaatccgccgcactcccagacgcccccgcgtcgacggcgc
	caacccgatccaagacacccgcgcaggggctggccagaaagct
	gcactttagcaccgcccccccaaaccccgacgcgccatggaccc
	cccgggtggccggctttaacaagcgcgtcttctgcgccgcggtcg
	ggcgcctggcggccatgcatgcccggatggcggcggtccagctc
	tgggacatgtcgcgtccgcgcacagacgaagacctcaacgaact
	ccttggcatcaccaccatccgcgtgacggtctgcgagggcaaaaa
	cctgcttcagcgcgccaacgagttggtgaatccagacgtggtgca
	ggacgtcgacgcggccacggcgactcgagggcgttctgcggcgt
	cgcgccccaccgagcgacctcgagccccagcccgctccgcttct
	cgccccagacggcccgtcgag
43	MTSRRSVKSGPREVPRDEYEDLYYTPSSG	Accession number: P10233
	MASPDSPPDTSRRGALFTQTRSRQRGEVR	Herpes Simplex virus 1 UL49/VP22
	FVQYDESDYALYGGSSSEDDEHPEVPRTR	protein sequence
	RPVSGAVLSGPGPARAPPPFTPAGSGGAG
	RTPTTAPRAPRTQRVATKAPAAPAAETTR
	GRKSAQPESAALPDAPASTAPTFTRSKTPA
	QGLARKLHFSTAPPNPDAPWTPRVAGFNK
	RVFCAAVGRLAAMHARMAAVQLWDFTM
	SRPRTDEDLNELLGITTIRVTVCEGKNLLQ
	RANELVNPDVVQDVDAATATRGRSAASR
	FTPTERPRAPARSASRPRRPVE

[0131] Melan-A mRNA Sequence

[0132] LOCUS NM—005511 1524 bu. mRNA PRI 14-OCT-2001

[0133] DEFINITION Homo sapiens melan-A (MLANA), mRNA.

[0134] ACCESSION NM—005511

[0135] VERSION NM—005511.1 GI:5031912

[0136]

/trans1ation =“MPREDAHFIYGYPKKGHGHSYTTAEEAAGIGILTVILGVLLLIGCWYCRRRN (SEQ ID NO.2)
GYRALMDKSLHVGTQCALTRRCPQEGFDHRDSKVSLQEKNCEPVVPNAPPAYEKLSAE
QSPPPYSP”

[0137]

ORIGIN
(SEQ ID NO. 35)
1    agcagacaga ggactctcat taaggaaggt gtcctgtgcc ctgaccctac aagatgccaa
61   gagaagatgc tcacttcatc tatggttacc ccaagaaggg gcacggccac tcttacacca
121  cggctgaaga ggccgctggg atcggcatcc tgacagtgat cctgggagtc ttactgctca
181  tcggctgttg gtattgtaga agacgaaatg gatacagagc cttgatggat aaaagtcttc
241  atgttggcac tcaatgtgcc ttaacaagaa gatgcccaca agaagggttt gatcatcggg
301  acagcaaagt gtctcttcaa gagaaaaact gtgaacctgt ggttcccaat gctccacctg
361  cttatgagaa actctctgca gaacagtcac caccacctta ttcaccttaa gagccagcga
421  gacacctgag acatgctgaa attatttctc tcacactttt gcttgaattt aatacagaca
481  tctaatgttc tcctttggaa tggtgtagga aaaatgcaag ccatctctaa taataagtca
541  gtgttaaaat tttagtaggt ccgctagcag tactaatcat gtgaggaaat gatgagaaat
601  attaaattgg gaaaactcca tcaataaatg ttgcaatgca tgatactatc tgtgccagag
661  gtaatgttag taaatccatg gtgttatttt ctgagagaca gaattcaagt gggtattctg
721  gggccatcca atttctcttt acttgaaatt tggctaataa caaactagtc aggttttcga
781  accttgaccg acatgaactg tacacagaat tgttccagta ctatggagtg ctcacaaagg
841  atacttttac aggttaagac aaagggttga ctggcctatt tatctgatca agaacatgtc
901  agcaatgtct ctttgtgctc taaaattcta ttatactaca ataatatatt gtaaagatcc
961  tatagctctt tttttttgag atggagtttc gcttttgttg cccaggctgg agtgcaatgg
1021 cgcgatcttg gctcaccata acctccgcct cccaggttca agcaattctc ctgccttagc
1081 ctcctgagta gctgggatta caggcgtgcg ccactatgcc tgactaattt tgtagtttta
1141 gtagagacgg ggtttctcca tgttggtcag gctggtctca aactcctgac ctcaggtgat
1201 ctgcccgcct cagcctccca aagtgctgga attacaggcg tgagccacca cgcctggctg
1261 gatcctatat cttaggtaag acatataacg cagtctaatt acatttcact tcaaggctca
1321 atgctattct aactaatgac aagtattttc tactaaacca gaaattggta gaaggattta
1381 aataagtaaa agctactatg tactgcctta gtgctgatgc ctgtgtactg ccttaaatgt
1441 acctatggca atttagctct cttgggttcc caaatccctc tcacaagaat gtgcagaaga
1501 aatcataaag gatcagagat tctg

[0138] Tyrosinase mRNA Sequence

[0139] LOCUS NM—000372 1964 bu. mRNA PRI 31-OCT-2000

[0140] DEFINITION Homo sapiens tyrosinase (oculocutaneous albinism IA) (TYR), mRNA.

[0141] ACCESSION NM—000372

[0142] VERSION NM—000372.1 GI:4507752

[0143]

/translation =“MLLAVLYCLLWSFQTSAGHFPRACVSSKNLMEKECCPPWSGDRS (SEQ ID NO.3)
PCGQLSGRGSCQNILLSNAPLGPQFPFTGVDDRESWPSVFYNRTCQCSGNFMGFNCGNC
KFGFWGPNCTERRLLVRRNWDLSAPEKDKFFAYLTLAKHTISSDYVTPIGTYGQMKNGS
TPMFNDINIYDLFVWMHYYVSMDALLGGSEIWRDIDFAHEAPAYLPWHRLFLLRWEQEI
QKLTGDENFTIPYWDWRDAEKCDICTDEYMGGQHPTNPNLLSPASFFSSWQIVCSRLEE
YNSHQSLCNGTPEGPLRRNPGNHDKSRTPRLPSSADVEFCLSLTQYESGSMDKAANFSFR
NTLEGFASPLTGIADASQSSMHNALHIYMNGTMSQVQGSANDPTFLLHHAFVDSIFEQWL
RRHRPLQEVYPEANAPIGHNRESYMVPFIPLYRNGDFFISSKDLGYDYSYLQDSDPDSFQ
DYIKSYLEQASRIWSWLLGAAMVGAVLTALLAGLVSLLCRHKRKQLP
EEKQPLLMEKEDYHSLYQSHL

[0144]

ORIGIN
(SEQ ID NO. 36)
1    atcactgtag tagtagctgg aaagagaaat ctgtgactcc aattagccag ttcctgcaga
61   ccttgtgagg actagaggaa gaatgctcct ggctgttttg tactgcctgc tgtggagttt
121  ccagacctcc gctggccatt tccctagagc ctgtgtctcc tctaagaacc tgatggagaa
181  ggaatgctgt ccaccgtgga gcggggacag gagtccctgt ggccagcttt caggcagagg
241  ttcctgtcag aatatccttc tgtccaatgc accacttggg cctcaatttc ccttcacagg
301  ggtggatgac cgggagtcgt ggccttccgt cttttataat aggacctgcc agtgctctgg
361  caacttcatg ggattcaact gtggaaactg caagtttggc ttttggggac caaactgcac
421  agagagacga ctcttggtga gaagaaacat cttcgatttg agtgccccag agaaggacaa
481  attttttgcc tacctcactt tagcaaagca taccatcagc tcagactatg tcatccccat
541  agggacctat ggccaaatga aaaatggatc aacacccatg tttaacgaca tcaatattta
601  tgacctcttt gtctggatgc attattatgt gtcaatggat gcactgcttg ggggatctga
661  aatctggaga gacattgatt ttgcccatga agcaccagct tttctgcctt ggcatagact
721  cttcttgttg cggtgggaac aagaaatcca gaagctgaca ggagatgaaa acttcactat
781  tccatattgg gactggcggg atgcagaaaa gtgtgacatt tgcacagatg agtacatggg
841  aggtcagcac cccacaaatc ctaacttact cagcccagca tcattcttct cctcttggca
901  gattgtctgt agccgattgg aggagtacaa cagccatcag tctttatgca atggaacgcc
961  cgagggacct ttacggcgta atcctggaaa ccatgacaaa tccagaaccc caaggctccc
1021 ctcttcagct gatgtagaat tttgcctgag tttgacccaa tatgaatctg gttccatgga
1081 taaagctgcc aatttcagct ttagaaatac actggaagga tttgctagtc cacttactgg
1141 gatagcggat gcctctcaaa gcagcatgca caatgccttg cacatctata tgaatggaac
1201 aatgtcccag gtacagggat ctgccaacga tcctatcttc cttcttcacc atgcatttgt
1261 tgacagtatt tttgagcagt ggctccgaag gcaccgtcct cttcaagaag tttatccaga
1321 agccaatgca cccattggac ataaccggga atcctacatg gttcctttta taccactgta
1381 cagaaatggt gatttcttta tttcatccaa agatctgggc tatgactata gctatctaca
1441 agattcagac ccagactctt ttcaagacta cattaagtcc tatttggaac aagcgagtcg
1501 gatctggtca tggctccttg gggcggcgat ggtaggggcc gtcctcactg ccctgctggc
1561 agggcttgtg agcttgctgt gtcgtcacaa gagaaagcag cttcctgaag aaaagcagcc
1621 actcctcatg gagaaagagg attaccacag cttgtatcag agccatttat aaaaggctta
1681 ggcaatagag tagggccaaa aagcctgacc tcactctaac tcaaagtaat gtccaggttc
1741 ccagagaata tctgctggta tttttctgta aagaccattt gcaaaattgt aacctaatac
1801 aaagtgtagc cttcttccaa ctcaggtaga acacacctgt ctttgtcttg ctgttttcac
1861 tcagcccttt taacattttc ccctaagccc atatgtctaa ggaaaggatg ctatttggta
1921 atgaggaact gttatttgta tgtgaattaa agtgctctta tttt

[0145] NY-ESO-1 mRNA Sequence

[0146] LOCUS HSU87459 752 bu. mRNA PRI 22-DEC-1999

[0147] DEFINITION Human autoimmunogenic cancer/testis antigen NY-ESO-1 mRNA, complete cds.

[0148] ACCESSION U87459

[0149] VERSION U87459.1 GI:1890098

[0150]

/translation =“MQAEGRGTGGSTGDADGPGGPGWDGPGGNAGGPGEAGATGGRGPRGAG (SEQ ID NO. 11)
AARASGPGGGAPRGPHGGAASGLNGCCRCGARGPESRLLEFYLAMPFATPMEAELARR
SLAQDAPPLPVPGVLLKEFTVSGNILTIRLTAADHRQLQLSISSCLQQLSLLM
WITQCFLPVFLAQPPSGQRR

[0151]

ORIGIN
(SEQ ID NO. 37)
1   atcctcgtgg gccctgacct tctctctgag agccgggcag aggctccgga gccatgcagg
61  ccgaaggccg gggcacaggg ggttcgacgg gcgatgctga tggcccagga ggccctggca
121 ttcctgatgg cccagggggc aatgctggcg gcccaggaga ggcgggtgcc acgggcggca
181 gaggtccccg gggcgcaggg gcagcaaggg cctcggggcc gggaggaggc gccccgcggg
241 gtccgcatgg cggcgcggct tcagggctga atggatgctg cagatgcggg gccagggggc
301 cggagagccg cctgcttgag ttctacctcg ccatgccttt cgcgacaccc atggaagcag
361 agctggcccg caggagcctg gcccaggatg ccccaccgct tcccgtgcca ggggtgcttc
421 tgaaggagtt cactgtgtcc ggcaacatac tgactatccg actgactgct gcagaccacc
481 gccaactgca gctctccatc agctcctgtc tccagcagct ttccctgttg atgtggatca
541 cgcagtgctt tctgcccgtg tttttggctc agcctccctc agggcagagg cgctaagccC
601 agcctggcgc cccttcctag gtcatgcctc ctcccctagg gaatggtccc agcacgagtg
661 gccagttcat tgtgggggcc tgattgtttg tcgctggagg aggacggctt acatgtttgt
721 ttctgtagaa aataaaactg agctacgaaa aa

[0152] PSMA cDNA Sequence

[0153] LOCUS NM—004476 2653 bu. mRNA PRI 01-NOV-2000

[0154] DEFINITION Homo sapiens folate hydrolase (prostate-specific membrane antigen) 1 (FOLH1), mRNA.

[0155] ACCESSION NM—004476

[0156] VERSION NM—004476.1 GI:4758397

[0157]

/trans1ation =“MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWIKSSNEAT (SEQ ID NO. 38)
NITPKHNMKAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVEL
AHYDVLLSYPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEG
DLVYVNYARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDP
ADYFAGVKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRRGIAEAVGL
PSIPVHPIGYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHS
TNEVTRIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGDPQSGAAVVHEIVRSFGTLKKE
GWRPRRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTP
LMYSLVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRL
GIASGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFE
LANSIVLPFDCRDYAVVLRKYADMYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFS
ERLQDFDKSNPIVLRMMNPQLMFLERAIUDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGI
YDALFDIESKVDPSKAWGEVKRQIYVAAFTVQAAAETLSEVA”

[0158]

ORIGIN
(SEQ ID NO. 39)
1    ctcaaaaggg gccggatttc cttctcctgg aggcagatgt tgcctctctc tctcgctcgg
61   attggttcag tgcactctag aaacactgct gtggtggaga aactggaccc caggtctgga
121  gcgaattcca gcctgcaggg ctgataagcg aggcattagt gagattgaga gagactttac
181  cccgccgtgg tggttggagg gcgcgcagta gagcagcagc acaggcgcgg gtcccgggag
241  gccggctctg ctcgcgccga gatgtggaat ctccttcacg aaaccgactc ggctgtggcc
301  accgcgcgcc gcccgcgctg gctgtgcgct ggggcgctgg tgctggcggg tggcttcttt
361  ctcctcggct tcctcttcgg gtggtttata aaatcctcca atgaagctac taacattact
421  ccaaagcata atatgaaagc atttttggat gaattgaaag ctgagaacat caagaagttc
481  ttatataatt ttacacagat accacattta gcaggaacag aacaaaactt tcagcttgca
541  aagcaaattc aatcccagtg gaaagaattt ggcctggatt ctgttgagct agcacattat
601  gatgtcctgt tgtcctaccc aaataagact catcccaact acatctcaat aattaatgaa
661  gatggaaatg agattttcaa cacatoatta tttgaaccac ctcctccagg atatgaaaat
721  gtttcggata ttgtaccacc tttcagtgct ttctctcctc aaggaatgcc agagggcgat
781  ctagtgtatg ttaactatgc acgaactgaa gacttcttta aattggaacg ggacatgaaa
841  atcaattgct ctgggaaaat tgtaattgcc agatatggga aagttttcag aggaaataag
901  gttaaaaatg cccagctggc aggggccaaa ggagtcattc tctactccga ccctgctgac
961  tactttgctc ctggggtgaa gtcctatcca gatggttgga atcttcctgg aggtggtgtc
1021 cagcgtggaa atatcctaaa tctgaatggt gcaggagacc ctctcacacc aggttaccca
1081 gcaaatgaat atgcttatag gcgtggaatt gcagaggctg ttggtcttcc aagtattcct
1141 gttcatccaa ttggatacta tgatgcacag aagctcctag aaaaaatggg tggctcagca
1201 ccaccagata gcagctggag aggaagtctc aaagtgccct acaatgttgg acctggcttt
1261 actggaaact tttctacaca aaaagtcaag atgcacatcc actctaccaa tgaagtgaca
1321 agaatttaca atgtgatagg tactctcaga ggagcagtgg aaccagacag atatgtcatt
1381 ctgggaggtc accgggactc atgggtgttt ggtggtattg accctcagag tggagcagct
1441 gttgttcatg aaattgtgag gagctttgga acactgaaaa aggaagggtg gagacctaga
1501 agaacaattt tgtttgcaag ctgggatgca gaagaatttg gtcttcttgg ttctactgag
1561 tgggcagagg agaattcaag actccttcaa gagcgtggcg tggcttatat taatgctgac
1621 tcatctatag aaggaaacta cactctgaga gttgattgta caccgctgat gtacagcttg
1681 gtacacaacc taacaaaaga gctgaaaagc cctgatgaag gctttgaagg caaatctctt
1741 tatgaaagtt ggactaaaaa aagtccttcc ccagagttca gtggcatgcc caggataagc
1801 aaattgggat ctggaaatga ttttgaggtg ttcttccaac gacttggaat tgcttcaggc
1861 agagcacggt atactaaaaa ttgggaaaca aacaaattca gcggctatcc actgtatcac
1921 agtgtctatg aaacatatga gttggtggaa aagttttatg atccaatgtt taaatatcac
1981 ctcactgtgg cccaggttcg aggagggatg gtgtttgagc tagccaattc catagtgctc
2041 ccttttgatt gtcgagatta tgctgtagtt ttaagaaagt atgctgacaa aatctacagt
2101 atttctatga aacatccaca ggaaatgaag acatacagtg tatcatttga ttcacttttt
2161 tctgcagtaa agaattttac agaaattgct tccaagttca gtgagagact ccaggacttt
2221 gacaaaagca acccaatagt attaagaatg atgaatgatc aactcatgtt tctggaaaga
2281 gcatttattg atccattagg gttaccagac aggccttttt ataggcatgt catctatgct
2341 ccaagcagcc acaacaagta tgcaggggag tcattcccag gaatttatga tgctctgttt
2401 gatattgaaa gcaaagtgga cccttccaag gcctggggag aagtgaagag acagatttat
2461 gttgcagcct tcacagtgca ggcagctgca gagactttga gtgaagtagc ctaagaggat
2521 tctttagaga atccgtattg aatttgtgtg gtatgtcact cagaaagaat cgtaatgggt
2581 atattgataa attttaaaat tggtatattt gaaataaagt tgaatattat atataaaaaa
2641 aaaaaaaaaa aaa

[0159] NM 003147 Homo Sapiens Synovial Sarcoma, X Breakpoint 2 (SSX2), mRNA

[0160] LOCUS NM—003147 766 bu. mRNA PRI 14-MAR-2001

[0161] DEFINITION Homo sapiens synovial sarcoma, X breakpoint 2 (SSX2), mRNA.

[0162] ACCESSION NM—003147

[0163] VERSION NM—003147.1 GI:10337582

[0164]

/translation =“MNGDDAFARRPTVGAQIPEKIQKAFDDTAKYFSKEEWEKMKASE SEQ ID NO. 40
KIFYVYMKRKYEANTKLGFKATLPPFMCNKRAEDFQGNDLDNDPNRGNQVERPQMTFG
RLQGISPKIMPKKPAEEGNDSEEVPEASGPQNDGKELCPPGKPTTSEKIHERSGPKRG
EHAWTHRLRERKQLVIYEEISDPEEDDE”

[0165]

SEQ ID NO 41
1   ctctctttcq attcttccat actcagagta cgcacggtct gattttctct ttggattctt
61  ccaaaatcag agtcagactg ctcccggtgc catgaacgga gacgacgcct ttgcaaggag
121 acccacggtt ggtgctcaaa taccagagaa gatccaaaag gccttcgatg atattgccaa
181 atacttctct aaggaagagt gggaaaagat gaaagcctcg gagaaaatCt tctatgtgta
241 tatgaagaga aagtatgagg ctatgactaa actaggtttc aaggccaccc tcccaccttt
301 catgtgtaat aaacgggccg aagacttcca ggggaatgat ttggataatg accctaaccg
361 tgggaatcag gttgaacgtc ctcagatgac tttcggcagg ctccagqgaa tctccccgaa
421 gatcatgccc aagaagccag cagaggaagg aaatgattcg gaggaagtgc cagaagcatc
481 tggcccacaa aatgatggga aagagctgtg ccccccggga aaaccaacta cctctgagaa
541 gattcacgag agatctggac ccaaaagggg ggaacatgcc tggacccaca gactgcgtga
601 gagaaaacag ctggtgattt atgaagagat cagcgaccct gaggaagatg acgagtaact
661 cccctcaggg atacgacaca tgcccatqat gagaagcaga acgtggtgac ctttcacgaa
721 catgggcatg gctgcggacc cctcgtcatc aggtgcatag caagtg

Claims

1. A method of identifying a polypeptide suitable for epitope liberation, the method comprising the steps of: identifying an epitope of interest; providing a substrate polypeptide sequence comprising the epitope, wherein the substrate polypeptide permits processing by a proteasome; contacting the substrate polypeptide with a composition comprising the proteasome, under conditions that support processing of the substrate polypeptide by the proteasome; and assaying for liberation of the epitope.

2. The method of claim 1, wherein the epitope is embedded in the substrate polypeptide.

3. The method of claim 1, wherein the epitope is a housekeeping epitope.

4. The method of claim 1, wherein the substrate polypeptide is a synthetic peptide.

5. The method of claim 1, wherein the substrate polypeptide is a fusion protein.

6. The method of claim 1, wherein the contacting step comprises immunization with the substrate polypeptide.

7. The method of claim 1, wherein the substrate polypeptide is encoded by a polynucleotide.

8. The method of claim 7, wherein the contacting step comprises immunization with a vector comprising the polynucleotide.

9. The method of claim 7, wherein the contacting step comprises transforming a cell with a vector comprising the polynucleotide.

10. The method of claim 1, wherein the proteasome processing takes place in vitro.

11. The method of claim 1, wherein the assaying step consists of a technique selected from the group consisting of mass spectrometry, N-terminal pool sequencing, and HPLC.

12. The method of claim 1, wherein the assaying step comprises a T cell target recognition assay.

13. The method of claim 1, wherein the substrate polypeptide further comprises an array of additional epitopes.

14. The method of claim 13, wherein the array comprises a housekeeping and an immune epitope.

15. The method of claim 1, wherein the substrate polypeptide further comprises an array of epitopes and epitope clusters.

16. The method of claim 1, wherein the proteasome is an immune proteasome.

17. A vector comprising a housekeeping epitope expression cassette, wherein the housekeeping epitope is derived from a target-associated antigen, and wherein the housekeeping epitope is liberatable from a translation product of the cassette by immunoproteasome processing.

18. The vector of claim 17, wherein the expression cassette encodes an array of two or more epitopes or at least one epitope and at least one epitope cluster.

19. The vector of claim 17, wherein the target-associated antigen is an antigen derived from or associated with a tumor or an intracellular parasite.

20. A method of activating a T cell comprising contacting the vector of claim 17 with an APC and contacting said APC with a T cell.

21. A substrate polypeptide comprising a housekeeping epitope wherein the housekeeping epitope can be liberated by immunoproteasome processing in a pAPC.