AU2004229458A1 - Improved heat shock protein-based vaccines and immunotherapies - Google Patents

Improved heat shock protein-based vaccines and immunotherapies Download PDF

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AU2004229458A1
AU2004229458A1 AU2004229458A AU2004229458A AU2004229458A1 AU 2004229458 A1 AU2004229458 A1 AU 2004229458A1 AU 2004229458 A AU2004229458 A AU 2004229458A AU 2004229458 A AU2004229458 A AU 2004229458A AU 2004229458 A1 AU2004229458 A1 AU 2004229458A1
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leu
arg
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Sofija Andjelic
Brian H. Barber
Jessica Baker Flechtner
Sunil Mehta
Kenya Prince-Cohane
Paul Slusarewicz
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Agenus Inc
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MOJAVE THERAPEUTICS Inc
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A—HUMAN NECESSITIES
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    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55516—Proteins; Peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K2039/6031—Proteins
    • A61K2039/6043—Heat shock proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
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    • A61K2039/622—Medicinal preparations containing antigens or antibodies characterised by the link between antigen and carrier non-covalent binding
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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Description

WO 2004/091493 PCT/US2004/010983 IMPROVED HEAT SHOCK PROTEIN-BASED VACCINES AND IMMUNOTHERAPIES CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to U.S. Patent Application Serial No. 5 10/776,521, filed February 12, 2004 and PCT Patent Application No. PCT/USO4/04340, filed February 13, 2004, both of which are incorporated herein by reference in their entireties. This application also claims priority under 35 U.S.C. § 119(e) to provisional applications serial no. 601462,469, filed April 11, 2003; serial no. 60/463,746, filed April 18, 2003; and serial no. 60/503,417, filed September 16, 2003, all three of which are 10 incorporated herein by reference in their entireties. INTRODUCTION [0002] The present invention relates to methods and compositions for inducing an immune response in a subject, wherein the subject is administered an effective amount of at least one or more defined hybrid antigens optionally in combination with one or more heat 15 shock proteins. These methods and compositions may be used in the treatment of infectious diseases and cancers. BACKGROUND OF THE INVENTION [0003] Heat shock proteins were originally observed to be expressed in increased amounts in mammalian cells which were exposed to sudden elevations of temperature, 20 while the expression of most cellular proteins is significantly reduced. It has since been determined that such proteins are produced in response to various types of stress, including glucose deprivation. As used herein, the term "heat shock protein" will be used to encompass both proteins that are expressly labeled as such as well as other stress proteins, including homologues of such proteins that are expressed constitutively (i.e., in the absence 25 of stressful conditions). Examples of heat shock proteins include BiP (also referred to as grp78), hsp70, hsc70, gp96 (grp94), hsp60, hsp40 and hsp90.
WO 2004/091493 PCT/US2004/010983 [0004] Heat shock proteins have the ability to bind other proteins in their non-native states, and in particular to bind nascent peptides emerging from ribosomes or extruded into the endoplasmic reticulum. Hendrick and Hartl, Ann. Rev. Biochem. 62:349-384 (1993); Hartl, Nature 381:571-580 (1996). Further, heat shock proteins have been shown to play an 5 important role in the proper folding and assembly of proteins in the cytosol, endoplasmic reticulum and mitochondria; in view of this function, they are referred to as "molecular chaperones." Frydman et al., Nature 370:111-117 (1994); Hendrick and Hartl, Ann. Rev. Biochem. 62:349-384 (1993); Hartl, Nature 381:571-580 (1996). [0005] For example, the protein BiP, a member of a class of heat shock proteins referred 10 to as the hsp70 family, has been found to bind to newly synthesized, unfolded p immunoglobulin heavy chain prior to its assembly with light chain in the endoplasmic reticulum. Hendershot et al., J. Cell Biol. 104:761-767 (1987). Another heat shock protein, gp96, is a member of the hsp90 family of stress proteins which localizes in the endoplasmic reticulum. Li and Srivastava, EMBO J. 12:3143-3151 (1993); Mazzarella and Green, J. 15 Biol. Chenzm. 262:8875-8883 (1987). It has been proposed that gp96 may assist in the assembly of multi-subunit proteins in the endoplasmic reticulum. Wiech et al., Nature 358:169-170 (1992). [0006] It has been observed that heat shock proteins prepared from tumors in experimental animals were able to induce immune responses in a tumor-specific manner; 20 that is to say, heat shock protein purified from a particular tumor could induce an immune response in an experimental animal which would inhibit the growth of the same tumor, but not other tumors. Srivastava and Maki, Curr. Topics Microbiol. 167:109-123 (1991). Genes encoding heat shock proteins have not been found to exhibit tumor-specific DNA polymorphism. Srivastava and Udono, Curr. Opin. Inmnunol. 6:728-732 (1994). High 25 resolution gel electrophoresis has indicated that gp96 may be heterogeneous at the molecular level. Feldweg and Srivastava, Int. J. Cancer 63: 310-314 (1995). Evidence suggests that the source of heterogeneity may be populations of small peptides adherent to the heat shock protein, which may number in the hundreds. Id. It has been proposed that a wide diversity of peptides adherent to tumor-synthesized heat shock proteins may render 30 such proteins capable of eliciting an immune response in subjects having diverse HLA 2 WO 2004/091493 PCT/US2004/010983 phenotypes, in contrast to more traditional immunogens which may be somewhat HLA restricted in their efficacy. Id. [0007] Nieland et al. (Proc. Natl. Acad. Sci. U.S.A. 93:6135-6139 (1996)) identified an antigenic peptide containing a cytotoxic T lymphocyte (CTL) vesicular stomatitis virus 5 (VSV) epitope bound to gp96 produced by VSV-infected cells. Neiland's methods precluded the identification of any additional peptides or other compounds which may also have bound to gp96, and were therefore unable to further characterize higher molecular weight material which was bound to gp96 and detected by high pressure liquid chromatography. 10 [0008] It has been reported that a synthetic peptide comprising multiple iterations of NANP (Asp Ala Asp Pro) malarial antigen, chemically cross-linked to glutaraldehyde-fixed mycobacterial hsp65 or hsp70, was capable of inducing antibody formation (i.e., a humoral response) in mice in the absence of any added adjuvant; a similar effect was observed using heat shock protein from the bacterium Escherichia coli. Del Guidice, Experientia 50:1061 15 1066 (1994); Barrios et al., Clin. Exp. Inmmunol. 98:224-228 (1994); Barrios et al., Eur. J. Immunol. 22:1365-1372 (1992). Cross-linking of synthetic peptide to heat shock protein and possibly glutaraldehyde fixation was required for antibody induction. Barrios et al., Clin. Exp. Immunol. 98:229-233. [0009] PCT/US96/13363 describes hybrid antigens comprising an antigenic domain and 20 a heat shock protein binding domain that, in a complex with a heat shock protein, induces immunological responses to antigens and are thus useful for treatment of cancer and infectious diseases. PCT/US98/22335 describes additional heat shock protein binding domains for similar uses, as well as the ability for hybrid antigen administered alone to induce an immune response. It has now been discovered that improvements in the peptide 25 linker present between the at least one antigenic domain and at least one heat shock protein binding domain in a hybrid antigen leads to an increase in biological activity. This increase is also found to provide an increase in inducing an immune response against the antigenic portion of the hybrid antigen. It is towards these improved peptide linkers, hybrid peptides containing them and their uses with and without heat shock protein, that the present 30 application is directed. 3 WO 2004/091493 PCT/US2004/010983 SUMMARY OF THE INVENTION [0010] The present invention relates to methods and compositions for inducing an immune response in a subject, wherein at least one defined hybrid antigen optionally in a 5 complex with a heat shock protein is administered to the subject. The hybrid antigen comprises at least one antigenic domain and at least one heat shock protein binding domain, and at least one peptide linker there between. Induction of an immune response to an antigen associated with a disease such as an infectious disease or tumor is useful for treatment of the disease. The antigenic or immunogenic domain of the hybrid antigen may 10 be an entire protein or peptide antigen, or may be only a portion of the selected antigen, for example a selected epitope of the antigen. The heat shock protein binding domain is a peptide that binds to a heat shock protein, preferably a peptide of 7-15 amino acids that binds to a heat shock protein, more preferably a hydrophobic peptide that binds to a heat shock protein, and most preferably a hydrophobic peptide of 7-15 amino acids that binds to 15 a heat shock protein. The linker has a sequence from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gln Leu Lys (QLK), Gin Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAI-AA 2
-AA
3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, 20 preferably V, more preferably S, and most preferably A; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. 25 [0011 ] The present invention provides for methods of administering such hybrid antigens alone, as well as heat shock protein/hybrid antigen compositions, the latter comprising (i) combining one or more heat shock protein with one or more hybrid antigens in vitro, under conditions wherein binding of hybrid antigen to heat shock protein occurs to form a hybrid antigen/heat shock protein complex; and (ii) administering the hybrid antigen, bound to heat 30 shock protein, in an effective amount to a subject in need of such treatment. 4 WO 2004/091493 PCT/US2004/010983 [0012] Alternatively, hybrid antigens optionally in combination with heat shock protein may be introduced into a subject by administering to the subject a nucleic acid encoding the hybrid antigen, optionally with nucleic acid encoding the heat shock protein. [0013] Thus, in a first aspect, the invention is directed to a hybrid antigen consisting 5 essentially of an antigenic domain of an infectious agent or tumor antigen, a binding domain that non-covalently binds to a heat shock protein, and a peptide linker separating the antigenic domain and the binding domain, and wherein the peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID 10 NO: 1003); Phe Arg (FR), Gin Leu Lys (QLK), Gin Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAl-AA 2
-AA
3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the 15 foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. [0014] In a second aspect, the invention is directed to a hybrid antigen consisting essentially of a plurality of antigenic domains of one or more infectious agents or one or more tumor antigens, at least one binding domain that non-covalently binds to a heat shock protein, and 20 at least one peptide linker separating the antigenic domains and the at least one binding domain, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys 25 Asn (KN); Arg Lys (RK); or AA 1
-AA
2
-AA
3 -leucine, wherein AAI is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, GIn Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg 30 Lys (FFRK; SEQ ID NO: 1000) is most preferred. In a particular embodiment, at least one of the antigenic domains in the aforementioned hybrid antigen is a T helper epitope. 5 WO 2004/091493 PCT/US2004/010983 [0015] In a third aspect, the invention is directed to a hybrid antigen comprising an antigenic domain of an infectious agent or tumor antigen and a binding domain that non covalently binds to a heat shock protein, and a peptide linker there between, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe 5 Arg Lys (FRK); Phe Arg Lys Amn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AA 1 -AAz-AA 3 -leucine, wherein AAI is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V 10 and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gn Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In a particular embodiment, the aforementioned hybrid antigen has a peptide linker separating the antigenic domain and the binding domain. 15 [0016] In a fourth aspect, the invention is directed to a hybrid antigen comprising a plurality of antigenic domains of one or more infectious agents or one or more tumor antigens and at least one binding domain that non-covalently binds to a heat shock protein, and at least one peptide linker there between, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn 20 (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gn Leu Lys (QLK), Gn Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO: 1001); Lys Asn (KN); Arg Lys (RK); or AAI-AA 2
-AA
3 -leucine, wherein AA, is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is 25 V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In a particular embodiment, at least one of the antigenic domains is a T helper epitope. [0017] In a fifth aspect, the invention is directed to a composition for inducing an 30 immune response to an infectious agent or tumor antigen comprising at least one hybrid antigen, the hybrid antigen comprising an antigenic domain of the infectious agent or tumor 6 WO 2004/091493 PCT/US2004/010983 antigen, a binding domain that non-covalently binds to a heat shock protein, and at least one peptide linker there between, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); 5 Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AA 1 -AAz-AA 3 -leucine, wherein AA, is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu 10 Lys (QLK), Arg Lys (RK) andAla Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In one embodiment, the composition comprises a plurality of hybrid antigens, and one of the hybrid antigens can comprise a T helper epitope. [0018] In a sixth aspect, the invention is directed to a composition for inducing an 15 immune response to an infectious agent or tumor antigen comprising at least one hybrid antigen, the hybrid antigen comprising a plurality of antigenic domains at least one of which is from the infectious agent or tumor antigen, at least one binding domain that non covalently binds to a heat shock protein, and at least one peptide linker there between, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID 20 NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO: 1001); Lys Asn (KN); Arg Lys (RK); or AAI-AAz-AA 3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, 25 more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. [0019] In a seventh aspect, the invention is directed to a composition for inducing an 30 immune response to an infectious agent or tumor antigen comprising at least one hybrid antigen, the hybrid antigen consisting essentially of an antigenic domain of the infectious 7 WO 2004/091493 PCT/US2004/010983 agent or tumor antigen, a binding domain that non-covalently binds to a heat shock protein, and a peptide linker separating the antigenic domain and the binding domain, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe 5 Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gin Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAi-AA 2
-AA
3 -leucine, wherein AAt is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S 10 and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In one embodiment, the aforementioned composition comprises a plurality of hybrid antigens. In another aspect, at least one of the plurality of hybrid antigens comprises a T helper epitope. 15 [0020] In an eighth aspect, the invention is directed to a composition for inducing an immune response to an infectious agent or tumor antigen comprising at least one hybrid antigen, the hybrid antigen consisting essentially of a plurality of antigenic domains at least one of which is from the infectious agent or tumor antigen, at least one binding domain that non-covalently binds to a heat shock protein, and at least one peptide linker separating the 20 antigenic domain and the binding domain, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gin Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AA 1
-AA
2
-AA
3 -leucine, 25 wherein AAI is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A; AAz is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. In one embodiment, at least one of the antigenic domains comprises a T helper epitope. [0021] In a ninth aspect, the invention is directed to a method for inducing an immune 30 response to an infectious agent or tumor antigen comprising administering to a subject a complex of a heat shock protein and a hybrid antigen comprising at least one antigenic 8 WO 2004/091493 PCT/US2004/010983 domain of the infectious agent or tumor antigen, at least one binding domain comprising a peptide that non-covalently binds to a heat shock protein, and a peptide linker there between; wherein the hybrid antigen and the heat shock protein are non-covalently bound, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID 5 NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gn Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO: 1001); Lys Asn (KN); Arg Lys (RK); or AA1-AA 2
-AA
3 -leucine, wherein AAI is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, 10 more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In one embodiment, the complex comprises a plurality of hybrid antigens. In an embodiment, at least one of the hybrid 15 antigens is a T helper epitope. In another embodiment, the hybrid antigen comprises a plurality of antigenic domains, and at least one of the antigenic domains can be a T helper epitope. In yet another embodiment wherein the complex comprises a plurality of hybrid antigens, at least one of the hybrid antigens comprises a plurality of antigenic domains. In another embodiment of this aspect of the invention, the heat shock protein is a hsp70. 20 [0022] In a tenth aspect, the invention is directed to a method for inducing an immune response to an infectious agent or tumor antigen comprising administering to a subject a complex of a heat shock protein and a hybrid antigen, the hybrid antigen consisting essentially of at least one antigenic domain of an infectious agent or tumor antigen, a binding domain that non-covalently binds to a heat shock protein, and a peptide linker 25 separating the antigenic domain and the binding domain, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Len Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AA 1
-AA
2
-AA
3 30 leucine, wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. 9 WO 2004/091493 PCT/US2004/010983 Among the foregoing, Gin Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In one embodiment, the complex comprises a plurality of hybrid antigens. In another embodiment, at least one of the hybrid antigens is a T helper epitope. In a further 5 embodiment, the hybrid antigen comprises a plurality of antigenic domains. In yet a further embodiment, at least one of the antigenic domains is a T helper epitope. In still yet another embodiment, the complex comprises a plurality of hybrid antigens, at least one of the hybrid antigens comprising a plurality of antigenic domains. In a preferred embodiment of this aspect, the heat shock protein is a hsp70. 10 [0023] In an eleventh aspect, the invention is directed to a method for inducing an immune response to an infectious agent or tumor antigen comprising administering to a subject at least one hybrid antigen comprising at least one antigenic domain of the infectious agent or tumor antigen, at least one binding domain comprising a peptide that non-covalently binds to a heat shock protein, and at least one peptide linker there between, 15 and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gin Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO: 1001); Lys Asn (KN); Arg Lys (RK); or AAl-AA2-AA 3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, 20 preferably V, more preferably S, and most preferably A; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000) is most preferred. In one embodiment, the complex 25 comprises a plurality of hybrid antigens. In another embodiment, at least one of the hybrid antigens is a T helper epitope. In another embodiment, the hybrid antigen comprises a plurality of antigenic domains. In a further embodiment, at least one of the antigenic domains is a T helper epitope. In yet a further embodiment, the complex comprises a plurality of hybrid antigens, at least one of the hybrid antigens comprising a plurality of 30 antigenic domains. In another embodiment of this aspect of the invention, a peptide linker separates the antigenic domain and the binding domain. 10 WO 2004/091493 PCT/US2004/010983 [0024] In a twelfth embodiment, the invention is directed to a method for inducing an immune response to an infectious agent or tumor antigen comprising administering to a subject at least one hybrid antigen, the hybrid antigen consisting essentially of at least one antigenic domain of an infectious agent or tumor antigen, a binding domain that non 5 covalently binds to a heat shock protein, and a peptide linker separating the antigenic domain and the binding domain, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gin Leu Lys (QLK), Gn Leu Glu (QLE), Ala Lys Val Leu 10 (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AA-AAz-AA 3 -leucine, wherein AAI is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID 15 NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In one embodiment, the complex comprises a plurality of hybrid antigens. In a further embodiment, at least one of the hybrid antigens is a T helper epitope. In another embodiment, the hybrid antigen comprises a plurality of antigenic domains. In yet another embodiment, at least one of the antigenic domains is a T helper epitope. In yet still a 20 further embodiment, the complex comprises a plurality of hybrid antigens, at least one of the hybrid antigens comprising a plurality of antigenic domains. [0025] In a thirteenth aspect, the invention is directed to a method for treating an infectious disease or cancer comprising administering to a subject a complex of a heat shock protein and a hybrid antigen comprising at least one antigenic domain of an infectious agent 25 or tumor antigen associated with the infectious disease or cancer, a binding domain comprising a peptide that non-covalently binds to a heat shock protein, and a peptide linker there between; and wherein the hybrid antigen and the heat shock protein are non-covalently bound, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg 30 Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gin Leu Lys (QLK), Gn Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAl-AA 2
-AA
3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, 11 WO 2004/091493 PCT/US2004/010983 preferably V, more preferably S, and most preferably A ; AAz is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg 5 Lys (FFRK; SEQ ID NO:1000) is most preferred. In one embodiment, the complex comprises a plurality of hybrid antigens. In another embodiment, at least one of the hybrid antigens is a T helper epitope. In yet another embodiment, the hybrid antigen comprises a plurality of antigenic domains. In still another embodiment, at least one of the antigenic domains is a T helper epitope. In yet still a further embodiment, the complex comprises a 10 plurality of hybrid antigens, at least one of the hybrid antigens comprising a plurality of antigenic domains. In an embodiment of this aspect of the invention, a peptide linker separates the antigenic domain and the binding domain. In a preferred embodiment of this aspect of the invention, the heat shock protein is a hsp70. [0026] In a fourteenth aspect, the invention is directed to a method for treating an 15 infectious disease or cancer comprising administering to a subject a complex of a heat shock protein and a hybrid antigen, the hybrid antigen consisting essentially of at least one antigenic domain of an infectious agent or tumor antigen associated with the infectious disease or cancer, at least one binding domain that non-covalently binds to a heat shock protein, and a peptide linker separating the antigenic domain and the binding domain, and 20 wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAl-AA 2
-AA
3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, 25 preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000) is most preferred. In one embodiment, the complex 30 comprises a plurality of hybrid antigens. In another aspect, at least one of the hybrid antigens is a T helper epitope. In yet another aspect, the hybrid antigen comprises a plurality of antigenic domains. In yet another aspect, at least one of the antigenic domains 12 WO 2004/091493 PCT/US2004/010983 is a T helper epitope. In a further aspect, the complex comprises a plurality of hybrid antigens, at least one of the hybrid antigens comprising a plurality of antigenic domains. In a preferred embodiment, the heat shock protein is a hsp70. [0027] In a fifteen aspect, the invention is directed to a method for treating an infectious 5 disease or cancer comprising administering to a subject at least one hybrid antigen comprising at least one antigenic domain of an infectious agent or tumor antigen associated with the infectious disease or cancer, a binding domain comprising a peptide that non covalently binds to a heat shock protein, and a peptide linker there between, and wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe 10 Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AA 1
-AA
2
-AA
3 -1eucine, wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A; AA 2 is K, V, or E, preferably E, more preferably V 15 and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gin Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO:1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO:1000) is most preferred. In one embodiment, the complex comprises a plurality of hybrid antigens. In another aspect, at least one of the hybrid antigens is a T helper epitope. 20 In yet another aspect, the hybrid antigen comprises a plurality of antigenic domains. In still a further aspect, at least one of the antigenic domains is a T helper epitope. In still yet another aspect, the complex comprises a plurality of hybrid antigens, at least one of the hybrid antigens comprising a plurality of antigenic domains. In one embodiment of this aspect of the invention, a peptide linker separates the antigenic domain and the binding 25 domain. [0028] In a sixteenth aspect, the invention is directed to a method for treating an infectious disease or cancer comprising administering to a subject at least one hybrid antigen, the hybrid antigen consisting essentially of at least one antigenic domain of an infectious agent or tumor antigen associated with an infectious disease or cancer, a binding 30 domain that non-covalently binds to a heat shock protein, and a peptide linker separating the antigenic domain and the binding domain, and wherein at least one peptide linker is from 13 WO 2004/091493 PCT/US2004/010983 among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gn Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAl-AA2-AA 3 -leucine, 5 wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gn Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000) is most preferred. 10 In one embodiment, the complex comprises a plurality of hybrid antigens. In another embodiment, at least one of the hybrid antigens is a T helper epitope. In yet another embodiment, the hybrid antigen comprises a plurality of antigenic domains. In still yet another embodiment, at least one of the antigenic domains is a T helper epitope. In another embodiment, the complex comprises a plurality of hybrid antigens, at least one of the hybrid 15 antigens comprising a plurality of antigenic domains. [0029] In a seventeenth aspect, the invention is directed to a peptide from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Giln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu 20 (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAI-AA 2
-AA
3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. [0030] In an eighteenth aspect, the invention is directed to an immunogenic polypeptide 25 comprising a plurality of antigenic domains, at least one heat shock protein binding domain and at leatst one peptide linker there between wherein at least one peptide linker is from among Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu 30 (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AA 1
-AA
2
-AA
3 -leucine, wherein AAI is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably 14 WO 2004/091493 PCT/US2004/010983 A; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000) is most preferred. 5 [0031] In a nineteenth aspect, the invention is directed to a polynucleotide encoding any of the hybrid antigens in the aforementioned first, second, third or fourth aspect. [0032] In a twentieth aspect, the invention is directed to a method of inducing an immune response to an infectious disease or cancer comprising administering to a subject a polynucleotide encoding a hybrid antigen comprising an antigenic domain of an infectious 10 agent or tumor antigen associated with the infectious disease or cancer, a heat shock protein binding domain, and a peptide linker there between from among Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gin Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys 15 Asn (KN); Arg Lys (RK); or AA 1
-AA
2
-AA
3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, more preferably S and most preferably V. Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg 20 Lys (FFRK; SEQ ID NO: 1000) is most preferred. [0033] In a twenty-first aspect, the invention is directed a method of inducing an immune response to an infectious disease or cancer comprising administering to a subject a polynucleotide encoding a hybrid antigen as mentioned above, and a polynucleotide encoding a heat shock protein. In a preferred embodiment, the encoded heat shock protein 25 is a hsp70. [0034] In any or all of the aforementioned aspects of the invention, the infectious disease antigen may be derived from an infectious agent such as a bacterium, virus, protozoan, mycoplasma, fungus, yeast, parasite, or prion, by way of non-limiting example. A cancer or tumor antigen associated with cancer may be derived from a sarcoma, a lymphoma, a 30 leukemia, or a carcinoma, a melanoma, carcinoma of the breast, carcinoma of the prostate, 15 WO 2004/091493 PCT/US2004/010983 ovarian carcinoma, carcinoma of the cervix, colon carcinoma, carcinoma of the lung, glioblastoma, or astrocytoma, by way of non-limiting examples. The antigenic domain of an infectious agent or cancer comprises an antigen derived from or associated with the infectious disease or tumor antigen, such that induction of an immune response to the 5 antigen of the infectious agent or cancer antigen, respectively, is useful for treating the corresponding infectious disease or cancer. [0035] This application claims priority under 35 U.S.C. § 119(e) to provisional applications serial no. 60/462,469, filed April 11, 2003; serial no. 60/463,746, filed April 18, 2003; and serial no. 60/503,417, filed September 16, 2003, all three of which arc 10 incorporated herein by reference in their entireties. BRIEF DESCRIPTION OF THE DRAWING [0036] Figure 1 shows the results of a tumor challenge study in which immunization using a hybrid antigen or complex of a hybrid antigen with a heat shock protein was 15 performed, followed seven days later by challenge with a tumor expressing the antigen. DETAILED DESCRIPTION OF THE INVENTION [0037] For purposes of clarity of description, and not by way of limitation, the detailed description is divided into the following subsections: 20 (i) hybrid antigens, (ii) heat shock proteins; and (iii) methods of administration. Hybrid Antigens [0038] A hybrid antigen, according to the invention comprises at least one antigenic 25 (immunogenic) domain, at least one heat shock protein-binding domain, and a peptide linker between at least two of these domains, wherein the peptide linker is among 16 WO 2004/091493 PCT/US2004/010983 Phe Phe Arg Lys (FFRK; SEQ ID NO:1000), Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); 5 Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001), 10 Lys Asn (KN); Arg Lys (RK); or AAl-AA 2
-AA
3 -leucine, wherein AA 1 is A, S, V, E, G, L, or K, preferably V, more preferably S, and most preferably A ; AA 2 is K, V, or E, preferably E, more preferably V and most preferably K; and AA 3 is V, S, F, K, A, E, or T, preferably F, 15 more preferably S and most preferably V. [0039] Among the foregoing, Gln Leu Lys (QLK), Arg Lys (RK) and Ala Lys Val Leu (AKVL; SEQ ID NO: 1001) are preferred, and Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000) is most preferred. [0040] Thus, the hybrid antigen serves at least two functions, namely (i) it contains an 20 epitope capable of inducing the desired immune response; and (ii) it is capable of physically binding to a heat shock protein. As will be noted below, such binding may occur in vivo such that administration of the hybrid antigen alone will induce the desired immune response and provide the desired therapeutic effect. [0041] The term "antigen" as used herein, refers to a compound which may be 25 composed of amino acids, carbohydrates, nucleic acids or lipids individually or in any combination. [0042] The term "hybrid antigen," as used herein, refers to a compound which binds to one or more heat shock proteins and which is representative of the immunogen toward which an immune response is desirably directed. For example, where the immunogen is an 30 influenza virus, the hybrid antigen may comprise a peptide fragment of the matrix protein of 17 WO 2004/091493 PCT/US2004/010983 the influenza virus. As used herein, the term "immunogen" is applied to the neoplastic cell, infected cell, pathogen, or component thereof, towards which an immune response is to be elicited, whereas the hybrid antigen comprises a portion of that immunogen which can provoke the desired response and which binds to one or more heat shock proteins. In 5 particular, the antigenic domain of the hybrid antigen is selected to elicit an immune response to a particular disease or pathogen, including peptides obtained from MHC molecules, mutated DNA gene products, and direct DNA products such as those obtained from tumor cells. [0043] While the invention maybe applied to any type of immunogen, immunogens of 10 particular interest are those associated with, derived from, or predicted to be associated with a neoplastic disease, including but not limited to a sarcoma, a lymphoma, a leukemia, or a carcinoma, and in particular, with melanoma, carcinoma of the breast, carcinoma of the prostate, ovarian carcinoma, carcinoma of the cervix, colon carcinoma, carcinoma of the lung, glioblastoma, astrocytoma, etc. Selections of melanoma antigens useful in hybrid 15 antigens of the present invention may be found, by way of non-limiting example, in PCT/US01/12449 (WO0178655), incorporated herein by reference in its entirety. Further, mutations of tumor suppressor gene products such as p53, or oncogene products such as ras may also provide hybrid antigens to be used according to the invention. [0044] In further embodiments, the immunogen may be associated with an infectious 20 disease, and, as such, may be a bacterium, virus, protozoan, mycoplasma, fungus, yeast, parasite, or prion. For example, but not by way of limitation, the immunogen may be a human papilloma virus (see below), a herpes virus such as herpes simplex or herpes zoster, a retrovirus such as human immunodeficiency virus 1 or 2, a hepatitis virus, an influenza virus, a rhinovirus, respiratory syncytial virus, cytomegalovirus, adenovirus, Mycoplasma 25 pneumoniae, a bacterium of the genus Sanlmonella, Staphylococcus, Streptococcus, Enterococcus, Clostridium, Escherichia, Klebsiella, Vibrio, Mycobacterium, amoeba, a malarial parasite, Trypanosomna cruzi, etc. [0045] Immunogens may be obtained by isolation directly from a neoplasm, an infected cell, a specimen from an infected subject, a cell culture, or an organism culture, or may be 30 synthesized by chemical or recombinant techniques. By way of non-limiting examples, suitable antigenic peptides, particularly for use in a hybrid antigen, for use against viruses, 18 WO 2004/091493 PCT/US2004/010983 bacteria and the like can be designed by searching through their sequences for MHC class I restricted peptide epitopes containing HLA binding sequences such as but not limited to HLA-A2 peptide binding sequences: Xaa(Leu/Met)XaaXaaXaa(Val/Ile/Leu/Thr)XaaXaa(Val/Leu) (SEQ ID NO:2), for 5 example, from viruses: Ser Gly Pro Ser Asn Thr Pro Pro Glu Ile (SEQ ID NO:31); Ser Gly Val Glu Asn Pro Gly Gly Tyr Cys Leu (SEQ ID NO:32); Lys Ala Val Tyr Asn Phe Ala Thr Cys Gly (SEQ ID NO:33); 10 Arg Pro Gln Ala Ser Gly Val Tyr Met (SEQ ID NO:34); Phe Gln Pro Gln Asn Gly Gln Phe Ile (SEQ ID NO:35); Ile Glu Gly Gly Trp Thr Gly Met Ile (SEQ ID NO:36); Thr Tyr Val Ser Val Ser Thr Ser Thr Leu (SEQ ID NO:37); Phe Glu Ala Asn Gly Asn Leu Ile (SEQ ID NO:38); 15 Ile Tyr Ser Thr Val Ala Ser Ser Leu (SEQ ID NO:39); Thr Tyr Gln Arg Thr Arg Ala Leu Val (SEQ ID NO:40); Cys Thr Glu Leu Lys Leu Ser Asp Tyr (SEQ ID NO:41); Ser Asp Tyr Glu Gly Arg Leu Ile (SEQ ID NO:42); Glu Glu Gly Ala Ile Val Gly Glu Ile (SEQ ID NO:43); 20 Val Ser Asp Gly Gly Pro Asn Leu Tyr (SEQ ID NO:44); Ala Ser Asn Glu Asn Met Glu Thr Met (SEQ ID NO:45); Ala Ser Asn Glu Asn Met Asp Ala Met (SEQ ID NO:46); Lys Leu Gly Glu Phe Tyr Asn Gln Met Met (SEQ ID NO:47); Leu Tyr Gln Asn Val Gly Thr Tyr Val (SEQ ID NO:48); 25 Thr Tyr Val Ser Val Gly Thr Ser Thr Leu (SEQ ID NO:49); Phe Glu Ser Thr Gly Asn Leu Ile (SEQ ID NO:50); Val Tyr Gln Ile Leu Ala Ile Tyr Ala (SEQ ID NO:51); Ile Tyr Ala Thr Val Ala Gly Ser Leu (SEQ ID NO:52); Gly Ile Leu Gly Phe Val Phe Thr Leu (SEQ ID NO:53); 30 Ile Leu Gly Phe Val Phe Thr Leu Thr Val (SEQ ID NO:54); Ile Leu Arg Gly Ser Val Ala His Lys (SEQ ID NO:55); Glu Asp Leu Arg Val Leu Ser Phe Ile (SEQ ID NO:56); 19 WO 2004/091493 PCT/US2004/010983 Glu Leu Arg Ser Arg Tyr Trp Ala Hle (SEQ ID NO:57); Ser Arg Tyr Trp Ala Ile Arg Thr Arg (SEQ ID NO:58); Lys Thr Gly Gly Pro Ile Tyr Lys Arg (SEQ ID NO:59); Phe Ala Pro Gly Asn Tyr Pro Ala Leu (SEQ ID NO:60); 5 Arg Arg Tyr Pro Asp Ala Val Tyr Leu (SEQ ID NO:61); Asp Pro Val Ile Asp Arg Leu Tyr Leu (SEQ ID NO:62); Ser Pro Gly Arg Ser Phe Ser Tyr Phe (SEQ ID NO:63); Tyr Pro Ala Leu Gly Leu His Glu Phe (SEQ ID NO:64); Thr Tyr Lys Asp Thr Val Gln Leu (SEQ ID NO:65); 10 Phe Tyr Asp Gly Phe Ser Lys Val Pro Leu (SEQ ID NO:66); Phe Ile Ala Gly Asn Ser Ala Tyr Glu Tyr Val (SEQ ID NO:67); Tyr Pro His Phe Met Pro Thr Asn Leu (SEQ ID NO:68); Ala Pro Thr Ala Gly Ala Phe Phe Phe (SEQ ID NO:69); Ser Thr Leu Pro Glu Thr Thr Val Val Arg Arg (SEQ ID NO:70); 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Val (SEQ ID NO:71); Trp Leu Ser Leu Leu Val Pro Phe Val (SEQ ID NO:72); Gly Leu Ser Pro Thr Val Trp Leu Ser Val (SEQ ID NO:73); Asp Leu Met Gly Tyr Ile Pro Leu Val (SEQ ID NO:74); Leu Met Gly Tyr Ile Pro Leu Val Gly Ala (SEQ ID NO:75); 20 Ala Ser Arg Cys Trp Val Ala Met (SEQ ID NO:76); Lys Leu Val Ala Leu Gly Ile Asn Ala Val (SEQ ID NO:77); Phe Leu Arg Gly Arg Ala Tyr Gly Leu (SEQ ID NO:78); Arg Arg Ile Tyr Asp Leu Ile Glu Leu (SEQ ID NO:79); Ile Val Thr Asp Phe Ser Val Ile Lys (SEQ ID NO:80); 25 Arg Arg Arg Trp Arg Arg Leu Thr Val (SEQ ID NO:81); Glu Glu Asn Leu Leu Asp Phe Val Arg Phe (SEQ ID NO:82); Cys Leu Gly Gly Leu Leu Thr Met Val (SEQ ID NO:83); Ser Ser Ile Glu Phe Ala Arg Leu (SEQ ID NO:84); Leu Tyr Arg Thr Phe Ala Gly Asn Pro Arg Ala (SEQ ID NO:85); 30 Asp Tyr Ala Thr Leu Gly Val Gly Val (SEQ ID NO:86); Leu Leu Leu Gly Thr Leu Asn Ile Val (SEQ ID NO:87); Leu Leu Met Gly Thr Leu Gly Ile Val (SEQ ID NO:88); 20 WO 2004/091493 PCT/US2004/010983 Thr Leu Gin Asp le Val Leu His Leu (SEQ ID NO:89); Gly Leu His Cys Tyr Glu Gin Len Val (SEQ ID NO:90); Pro Leu Lys Gln His Phe Gin Ile Val (SEQ ID NO:91); Arg Len Val Thr Leu Lys Asp Ile Val (SEQ ID NO:92); 5 Arg Ala His Tyr An Ile Val Thr Phe (SEQ ID NO:93); Leu Leu Phe Gly Tyr Pro Val Tyr Val (SEQ ID NO:94); Ser Ala Ile Asn Asn Tyr Ala Gln Lys Leu (SEQ ID NO:95); His Gin Ala Ile Ser Pro Arg Thr Leu (SEQ ID NO:96); Gln Met Val His Gln Ala Ile Ser Pro Arg Thr Leu (SEQ ID NO:97); 10 Cys Lys Gly Val Asn Lys Glu Tyr Leu (SEQ ID NO:98); Gln Gly Ile Asn Asn Leu Asp Asn Leu (SEQ ID NO:99); Asn Asn Leu Asp Asn Lou Arg Asp Tyr (SEQ ID NO:100); Ser Glu Phe Leu Leu Glu Lys Arg lie (SEQ ID NO:101); Ser Tyr Ile Gly Ser Ile Asn Asn Ile (SEQ ID NO: 102); 15 Ile Leu Gly Asn Lys Ile Val Arg Met Tyr (SEQ ID NO:103); Arg Leu Arg Pro Gly Gly Lys Lys Lys (SEQ ID NO: 104); Glu Ile Lys Asp Thr Lys Glu Ala Leu (SEQ ID NO: 105); Gly Glu Ile Tyr Lys Arg Trp Ile Ile (SEQ ID NO: 106); Glu He Tyr Lys Arg Trp Ile Ile Leu (SEQ ID NO:107); 20 Arg Tyr Leu Lys Asp Gin Gin Leu Leu (SEQ ID NO:108); Arg Gly Pro Gly Arg Ala Phe Val Thr Ile (SEQ ID NO: 109); lie Val Gly Leu Asn Lys Ile Val Arg (SEQ ID NO:110); Thr Val Tyr Tyr Gly Val Pro Val Trp Lys (SEQ ID NO: 111); Arg Leu Arg Asp Leu Leu Leu lie Val Thr Arg (SEQ ID NO: 112); 25 Lys Arg Trp Ile 1le Leu Gly Leu Asn Lys (SEQ ID NO: 113); Ser Phe Asn Cys Gly Gly Glu Phe Phe (SEQ ID NO: 114); Gly Arg Ala Phe Val Thr Ile Gly Lys (SEQ ID NO: 115); Thr Pro Gly Pro Gly Val Arg Tyr Pro Leu (SEQ ID NO:116); Gin Val Pro Leu Arg Pro Met Thr Tyr Lys (SEQ ID NO:117); 30 Thr Glu Met Glu Lys Glu Gly Lys Ile (SEQ ID NO:118); Ile Leu Lys Glu Pro Val His Gly Val (SEQ ID NO:119); Val Glu Ala Glu Ile Ala His Gln Ile (SEQ ID NO:120); 21 WO 2004/091493 PCT/US2004/010983 Arg Gly Tyr Val Tyr Gln Gly Leu (SEQ ID NO:121); Tyr Ser Gly Tyr Ile Phe Arg Asp Leu (SEQ ID NO:122); Val Gly Pro Val Phe Pro Pro Gly Met (SEQ ID NO:123); lie Ile Tyr Arg Phe Leu Leu Ile (SEQ ID NO:124); 5 from bacteria: Lys Tyr Gly Val Ser Val Gln Asp Ile (SEQ ID NO:125); Ile Gin Val Gly Asn Thr Arg Thr Ile (SEQ ID NO: 126); Thr Pro His Pro Ala Arg Ile Gly Leu (SEQ ID NO:127); from parasites: 10 Ser Tyr Ile Pro Ser Ala Glu Lys Ile (SEQ ID NO:128); Lys Pro Lys Asp Glu Leu Asp Tyr (SEQ ID NO:129); Lys Ser Lys Asp Glu Leu Asp Tyr (SEQ ID NO:130); Lys Pro Asn Asp Lys Ser Leu Tyr (SEQ ID NO:131); Lys Tyr Leu Lys Lys Ile Lys Asn Ser Leu (SEQ ID NO:132); 15 Tyr Glu Asn Asp Ile Glu Lys Lys Ile (SEQ ID NO:133); Asn Tyr Asp Asn Ala Gly Thr Asn Leu (SEQ ID NO: 134); Asp Glu Leu Asp Tyr Glu Asn Asp Ile (SEQ ID NO:135); Ser Tyr Val Pro Ser Ala Glu Gln Ile (SEQ ID NO:136); from cancers: 20 Phe Glu Gin Asn Thr Ala Gin Pro(SEQ ID NO: 137); Phe Glu Gln Asn Thr Ala Gin Ala (SEQ ID NO:138); Glu Ala Asp Pro Thr Gly His Set Tyr (SEQ ID NO:139); Glu Val Asp Pro lie Gly His Leu Tyr (SEQ ID NO:140); Ala Ala Gly Ile Gly Ile Leu Thr Val (SEQ ID NO:141); 25 Tyr Leu Glu Pro Gly Pro Val Thr Ala (SEQ ID NO: 142); Ile Leu Asp Gly Thr Ala Thr Leu Arg Leu (SEQ ID NO:143); Met Leu Leu Ala Leu Leu Tyr Cys Leu (SEQ ID NO:144); Tyr Met Asn Gly Thr Met Set Gin Val (SEQ ID NO:145); Leu Pro Tyr Leu Gly Trp Leu Val Phe (SEQ ID NO:146); 30 Phe Gly Pro Tyr Lys Leu Asn Arg Leu (SEQ ID NO: 147); Lys Ser Pro Trp Phe Thr Thr Leu (SEQ ID NO:148); Gly Pro Pro His Ser Asn Asn Phe Gly Tyr (SEQ ID NO:149); and 22 WO 2004/091493 PCT/US2004/010983 Ile Ser Thr Gln Asn His Arg Ala Leu (SEQ ID NO:150) (Rammensee et al., Immunogenetics 41:178-223 (1995)), Xaa(Leu/Met)XaaXaaXaaXaaXaaXaaVal (SEQ ID NO:3) (Tarpey et al., Inmunology 81:222-227 (1994)), 5 Xaa(Val/Gln)XaaXaaXaaXaaXaaXaaLeu (SEQ ID NO:28), for example, from virus: Tyr Gly Ile Leu Gly Lys Val Phe Thr Leu (SEQ ID NO: 151); Ser Leu Tyr Asn Thr Val Ala Thr Leu (SEQ ID NO: 152); (Barouch et al., J. Exp. Med. 182:1847-1856 (1995)). 10 [0046] The foregoing epitopes are merely exemplary of selections available associated with various infectious diseases and cancer, and are provided without any intention whatsoever to be limiting. [0047] It may also be desirable to consider the type of immune response which is desired. For example, under certain circumstances, a humoral immune response may be 15 appropriate. In other cases, and indeed where an immune response directed toward neoplastic cells or infected cells is sought to be elicited, a cellular immune response is particularly desirable. Accordingly, particular epitopes associated with the activation of B cells, T helper cells, or cytotoxic T cells may be identified and selected for incorporation into the hybrid antigen. 20 [0048] It may also be desirable to utilize hybrid antigen associated with an autoimmune disease or allergy. Such a hybrid antigen may be administered, together with one or more heat shock proteins, in an amount sufficient to be tolerogenic or to inhibit a pre-existing immune response to the hybrid antigen in a subject. The amount of heat shock protein required to inhibit the immune response is expected to be substantially greater than the 25 amount required for stimulation. [0049] Although the size of hybrid antigen may vary depending upon the heat shock protein used, in non-limiting embodiments of the invention, the hybrid antigen may be the size of a peptide having between 10 and 500 amino acid residues, and preferably be the size of a peptide having between 14 and 100, most preferably 18 and 50 amino acid residues. 30 As such, it may be desirable to produce a fragment of an immunogen to serve as the 23 WO 2004/091493 PCT/US2004/010983 antigenic domain of a hybrid antigen, or, alternatively, to synthesize a hybrid antigen by chemical or recombinant DNA methods. [0050] Based on the foregoing considerations, a hybrid antigen may be prepared, and then tested for its ability to bind to heat shock protein. In some instances, binding of hybrid 5 antigen to a particular heat shock protein may be facilitated by the presence of at least one other protein, which may be a heat shock protein. [0051] For example, binding of hybrid antigen to a heat shock protein may be evaluated by labeling the hybrid antigen with a detectable label, such as a radioactive, fluorescent, enzymatic or pigmented label, combining the hybrid antigen with heat shock protein under 10 conditions which would be expected to permit binding to occur, and then isolating the heat shock protein while removing any unbound hybrid antigen, and determining whether any labeled hybrid antigen had adhered to the heat shock protein. As a specific example, and not by way of limitation, the ability of a hybrid antigen to bind to the heat shock protein BiP may be evaluated by combining 2 pg BiP with up to about 1150 pmole of radioactively 15 labeled hybrid antigen in buffer containing 50 mM Tris HCI (pH 7.5), 200 mM NaC1, and 1 mM Na 2 EDTA, in a final volume of 50 p l, for 30 minutes at 37 degrees Centigrade. Unbound hybrid antigen may then be removed from bound BiP-hybrid antigen by centrifugation at 100g by desalting through a 1 ml Sephadex-G column for 2 minutes. Penefsky, J Biol. Chem. 252:2891 (1977). To prevent binding to the resin, columns may 20 first be treated with 100 pl of bovine serum albumin in the same buffer and centrifuged as above. Bound hybrid antigen may then be quantitated by liquid scintillation counting. See Flynn et al., Science 245:385-390 (1989). [0052] Because ATP hydrolysis drives the release of peptides from many known heat shock proteins, the amount of ATPase activity may often be used to quantitate the amount 25 of hybrid antigen binding to heat shock protein. An example of how such an assay may be performed is set forth in Flynn et al., Science 245:385-390 (1989). [0053] The heat shock protein-binding domain is selected so that the hybrid antigen will bind in vitro or in vivo to a heat shock protein such as BiP, hsp70, gp96, or hsp90, or a member of the foregoing heat shock protein families, alone or in combination with 30 accessory heat shock proteins such as hsp40, or hsp60. 24 WO 2004/091493 PCT/US2004/010983 [0054] Non-limiting examples ofpeptides which fulfill this criterion may be identified by panning libraries of antigens known to bind well to one or more heat shock proteins as described in Blond-Elguindi et al., Cell 75:717-728 (1993): Leu Phe Trp Pro Phe Glu Trp Ile (SEQ ID NO:153); 5 Asp Gly Val Gly Ser Phe le Gly (SEQ ID NO: 154); Glu Ser Leu Trp Asn Pro Gln Cys (SEQ ID NO:155); Leu His Phe Asp Val Leu Trp Arg (SEQ ID NO: 156); Cys His Leu Lys Met Val Pro Trp (SEQ ID NO:157); Asn Ser Val Leu Val Cys Glu Leu (SEQ ID NO:158); 10 Asp Arg Gly His Ser Thr Tyr Ser (SEQ ID NO:159); Asp Val Trp Gly Tip Val Thr Trp (SEQ ID NO:160); Ile Gln Phe Arg Val Glu Leu Phe (SEQ ID NO:161); Leu Trp Leu Glu Leu Ser Leu Ser (SEQ ID NO: 162); Val Gly Ile Cys Ala Leu Phe Gly (SEQ ID NO:163); 15 Pro Tyr Pro Ser Gly Leu Asp Ser (SEQ ID NO:164); Phe Trp Gly Val Leu Pro Tyr Pro (SEQ ID NO:165); Phe Thr His Gly Ile Ser Leu Tyr (SEQ ID NO:166); Asn His Ser Phe Gly Gly Ser Thr (SEQ ID NO:167); Val Asp Tyr Val Tyr Phe His His (SEQ ID NO: 168); 20 Phe Leu Asp Ile lie Gly Tyr Gly (SEQ ID NO:169); Trp Asp Asp Leu Leu His Gly Arg (SEQ ID NO:170); Leu Arg Leu Leu Gly Thr Leu Asn (SEQ ID NO:171); Phe Glu Gln His Asn Gin Glu Pro (SEQ ID NO: 172); Phe Val Gly Thr Val Thr Trp Ser (SEQ ID NO:173); 25 Leu Trp Ala Leu Thr Tyr Arg Gly (SEQ ID NO: 174); Ser Trp Gly Ser Asn Gly Gly Phe (SEQ ID NO:175); Asp Met Trp Arg Arg Ala Val Gln (SEQ ID NO:176); Cys Arg Val Ile Tyr His Ala Thr (SEQ ID NO:177); Met Val Val Ala Arg Cys Gly His (SEQ ID NO:178); 30 His Met Trp Ile Asn Trp Val Gin (SEQ ID NO: 179); Cys Ala Gly Arg Cys Phe Gly Tyr (SEQ ID NO:180); Cys Thr His Val Leu Ala Tyr Ser (SEQ ID NO:181); 25 WO 2004/091493 PCT/US2004/010983 Ser Trp Met Pro Trp Leu Thr Met (SEQ ID NO: 182); Leu Glu Trp Cys Ile Tip Arg Tyr (SEQ ID NO:183); Cys Leu Ala Cys Ile Ile His Ser (SEQ ID NO:184); Phe Trp Phe Pro Trp Asp Arg Ser (SEQ ID NO:185); 5 Trp Arg Thr Gly Val Phe His Gly (SEQ ID NO: 186); Met His Leu Arg Val Ala Asp Arg (SEQ ID NO: 187); Ala Leu Asp Leu Tyr Leu Tyr Val (SEQ ID NO:188); Phe Phe Trp Phe Thr Leu Lys Glu (SEQ ID NO:189); Leu Ser Phe Ala Gly Trp Gly Val (SEQ ID NO:190); 10 Met Met Met Leu Gly Arg Ala Pro (SEQ ID NO:191); Trp Set Phe Tyr Thr Tip Leu Asn (SEQ ID NO:192); Phe Val Trp Met Arg Trp Ile Asp (SEQ ID NO:193); Met Gln Val Asn Thr Pro Asp Asn (SEQ ID NO:194); Phe Trp Gly Trp Leu Ile Pro Trp (SEQ ID NO:195); 15 Trp Gly Trp Val Trp Trp Asp (SEQ ID NO:196); Trp Ile Phe Pro Trp Ile Gln Leu (SEQ ID NO:197); Trp Met Phe Asn Trp Pro Tip Tyr (SEQ ID NO:198); Met Asn Met Ile Val Leu Asp Lys (SEQ ID NO: 199); Phe Trp Gly Tip Pro Gly Trp Set (SEQ ID NO:200); 20 Trp Leu Ile Arg Val Gly Thr Ala (SEQ ID NO:201); Gly Leu Leu Thr His Leu Ile Trp (SEQ ID NO:202); Leu Trp Trp Leu Asn Val His Gly (SEQ ID NO:203); Trp Trp Trp Ile Asn Asp Glu Ser (SEQ ID NO:204); Ala Asn Pro Set Leu Ala Thr Tyr (SEQ ID NO:205); 25 Trp Leu Gln Gly Trp Trp Gly Trp (SEQ ID NO:206); Met Met Pro Val Thr Ser Phe Arg (SEQ ID NO:207); Gly
T
rp Met Asp
T
rp Trp Tyr Tyr (SEQ ID NO:208); Leu Ala Ser Met Arg Asn Ser Met (SEQ ID NO:209); Asp Leu Met Arg Trp Leu Gly Leu (SEQ ID NO:210); 30 Tyr Phe Tyr Ala Trp Trp Leu Asp (SEQ ID NO:211); Leu Gly His Leu Trp Thr Gin Val (SEQ ID NO:212); Leu Trp Trp Arg Asp Val Met Ala (SEQ ID NO:213); 26 WO 2004/091493 PCT/US2004/010983 Phe Ile Trp Tirp Ala Pro Leu Ala (SEQ ID NO:214); Gly Ser Val Gly Gly Gly Val Val (SEQ ID NO:215); Asp Ser His Asp Asp Trp Arg Met (SEQ ID NO:216); Phe Tirp Arg Phe Asp Tyr Tyr Phe (SEQ ID NO:217); 5 Trp Thr Trp Trp Glu Trp Leu Ala (SEQ ID NO:218); Trp Leu Trp Asp Trp Ile Val Val (SEQ ID NO:219); Gly Trp Thr Trp Phe Phe Asp Met (SEQ ID NO:220); Ala Trp Trp Gln His Phe Ile Val (SEQ ID NO:221); Leu Trp Trp Asp Ile Ile Thr Gly (SEQ ID NO:222); 10 Phe Thr Tyr Gly Ser Arg Tirp Leu (SEQ ID NO:223); Phe Ser Leu Trp Pro Leu Ala Trp (SEQ ID NO:224); Gly Ile Ile Leu Gly Tyr Asn Val (SEQ ID NO:225); Ser Trp Met Thr Trp Ile G1u His (SEQ ID NO:226); Gly Trp Trp Val Thr Trp Pro Tirp (SEQ ID NO:227); 15 Val Val Ser Pro Tirp Tirp Leu Gly (SEQ ID NO:228); Asn Val Leu Ser Arg Gly Phe Ser (SEQ ID NO:229); Ser Phe Glu Ser Leu Gly Gly Leu (SEQ ID NO:230); Ile Thr Lys Gly Ser Ser Phe Pro (SEQ ID NO:231); Leu Asp Trp Ala Arg Lys Leu Arg (SEQ ID NO:232); 20 Thr Ala Trp Asn Leu Leu Gly Tyr (SEQ ID NO:233); Phe Gly Gln Gly Ile Lys His Val (SEQ ID NO:234); Asp Val Val Trp Gin Arg Leu Leu (SEQ ID NO:235); Tyr Val Asp Arg Phe Ile Gly Trp (SEQ ID NO:236); Lys Met Ala Arg Pro Glu Gly Asn (SEQ ID NO:237); 25 Leu Gly Arg Trp Gly His Glu Ser (SEQ ID NO:238); Ser lie Trp Ser Leu Leu Val Leu (SEQ ID NO:239); Val Trp Leu Asp Leu Leu Leu Ser (SEQ ID NO:240); Tyr Leu Thr Asp Ser Leu Phe Gly (SEQ ID NO:241); Thr Trp Trp Pro Ser Ile Thr Trp (SEQ ID NO:242); 30 Tyr Gly Leu Trp Trp Phe Pro Trp (SEQ ID NO:243); Phe Ser Pro Ala Asp Thr Arg Tyr (SEQ ID NO:244); Cys Asn Arg Leu Gln Ile Asp Cys (SEQ ID NO:245); 27 WO 2004/091493 PCT/US2004/010983 Ser Leu Val Ala Ala Arg Asn Leu (SEQ ID NO:246); Phe Thr Ile His Asn Val Ala Val (SEQ ID NO:247); Met Gly Pro Leu Gly Pro Leu Leu (SEQ ID NO:248); Arg Gln Leu Ser Glu Leu Phe Val (SEQ ID NO:249); 5 Arg Val Val Cys Gin Ala Leu Leu (SEQ ID NO:250); Trp Pro His Leu Trp Trp Leu Asp (SEQ ID NO:251); Trp Met Asp Trp Val Trp His Thr (SEQ ID NO:252); Trp Trp Gly Tyr Leu Ile Cys Gin (SEQ ID NO:253); Phe Arg Gly Leu Set Glu Gly Pro (SEQ ID NO:254); 10 Set Trp Phe Asp Trp Leu Val Ala (SEQ ID NO:255); Val Val Met Trp Tyr Ser Val Asp (SEQ ID NO:256); Trp Gly Trp Set Leu Ala Thr (SEQ ID NO:257); Leu Gly Trp Phe Asp Arg Phe Phe (SEQ ID NO:258); Ala Trp Trp Trp Pro Thr Tyr Val (SEQ ID NO:259); 15 Gly Phe Leu Ser Ser Trp Phe Leu (SEQ ID NO:260); Gly Val Ile Asn Cys Ala Gly Thr (SEQ ID NO:261); Val Cys Ala Arg Ala Ala His Leu (SEQ ID NO:262); Qly Asn Set Tyr Gly Asp Gly Gly (SEQ ID NO:263); Gly Phe Leu Ser Ser Trp Phe Leu (SEQ ID NO:264); 20 Phe Asp Gln Pro Gly Arg Phe Leu (SEQ ID NO:265); Arg Set His Ala Thr Gly Val Val (SEQ ID NO:266);. Gly Tyr Trp Ala Met Met Set Trp (SEQ ID NO:267); Cys His Ser Met Trp Asp Gly Leu (SEQ ID NO:268); Phe Ile Trp Arg Gly Trp Pro His (SEQ ID NO:269); 25 Leu Set Phe Leu Gly Gly Arg Leu (SEQ ID NO:270); Phe Set Gly Val Arg Gln Pro Asn (SEQ ID NO:271); Trp Gly Trp Met Pro Phe Tyr Tyr (SEQ ID NO:272); Phe Thr Arg Pro Ala Val Val Asp (SEQ ID NO:273); Asp Leu Trp Thr Trp Leu Gly Leu (SEQ ID NO40:274); 30 Cys Asp Thr Ala Ala Val Ala Asp (SEQ ID NO:275); Tip Trp Val Lys His His Met Leu (SEQ ID NO:276); Ile Ala Phe Leu Arg Asp Asn Arg (SEQ ID NO:277); 28 WO 2004/091493 PCT/US2004/010983 Leu Ala Arg Pro Asp His Tyr Ser (SEQ ID NO:278); Met Glu Ser Lys Arg Trp Thr Val (SEQ ID NO:279); Met Ile Leu Lys Gly Tyr Ser Arg (SEQ ID NO:280); Ala Pro Ser Asp Tyr Asp Glu Ser (SEQ ID NO:281); 5 His Trp Leu Arg Ser Lys Arg Thr (SEQ ID NO:282); Gly Ala Arg Val Trp Asn Tyr Gln (SEQ ID NO:283); Leu Ser Asn Trp Asn Met Arg Leu (SEQ ID NO:284); Cys Gly Ala Ala Gin Gin Gly Met (SEQ ID NO:285); and Gly Ser Ser Met Val Val Gln Arg (SEQ ID NO:286). Using this technique, Blond-Elguindi 10 have concluded that the heat shock protein BiP recognizes polypeptides that contain a heptameric region having the sequence Hy(Trp/X)HyXHyXHy (SEQ ID NO: 29) where Hy represents a hydrophobic amino acid residue, particularly tryptophan, leucine or phenylalanine (SEQ ID NO:30), and X is any amino acid. High affinity heat-shock protein 15 binding sequences incorporating this motif include: His Trp Asp Phe Ala Trp Pro Trp (SEQ ID NO:1); and Phe Trp Gly Leu Trp Pro Trp Glu (SEQ ID NO:4). [0055] Other heat shock protein binding motifs have also been identified. For example, Auger et al., Nature Medicine 2:306-310 (1996) have identified two pentapeptide binding 20 motifs Gln Lys Arg Ala Ala (SEQ ID NO:5) and Arg Arg Arg Ala Ala (SEQ ID NO:6) in HLA-DR types associated with rheumatoid arthritis which bind to heat shock proteins. Heat shock binding motifs have also been identified as consisting of seven to fifteen residue 25 long peptides which are enriched in hydrophobic amino acids. Lys Arg Gln Ile Tyr Asp Leu Glu Met Asn Arg Leu Gly Lys (SEQ ID NO:287); Leu Ser Ser Leu Phe Arg Pro Lys Arg Arg Pro Ile Tyr Lys Ser (SEQ ID NO:288); Lys Lea Ile Gly Val Leu Ser Ser Leu Phe Arg Pro Lys (SEQ ID NO:289); Arg Arg Pro Ile Tyr Lys Ser Asp Val Gly Met Ala His Phe Arg (SEQ ID NO:290); 30 Cys Lys Ile Gln Ser Thr Pro Val Lys Gin Ser (SEQ ID NO:291); Tyr His Cys Asp Gly Phe Gln Asn Glu (SEQ ID NO:292); Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys (SEQ ID NO:293); 29 WO 2004/091493 PCT/US2004/010983 Ser Asn Gly Ser Leu Gin Cys Arg Ile Cys (SEQ ID NO:294) (Flynn et al., Science 245: 385-390 (1989)), [0056] Moreover, other heat shock protein binding peptides include: 5 Gly Lys Trp Val Tyr Ile (SEQ ID NO:295); Ala Lys Arg Glu Thr Lys (SEQ ID NO:296); Lys Trp Val His Leu Phe (SEQ ID NO:297); Arg Leu Val Leu Val Leu (SEQ ID NO:298); Trp Lys Trp Gly Ile Tyr (SEQ ID NO:299); 10 Ser Ser His Ala Ser Ala (SEQ ID NO:300); Trp Gly Pro Trp Ser Phe (SEQ ID NO:301); Ala Ile Pro Gly Lys Val (SEQ ID NO:302); Arg Val His Asp Pro Ala (SEQ ID NO:303); Arg Ser Val Ser Ser Phe (SEQ ID NO:304); 15 Leu Gly Thr Arg Lys Gly (SEQ ID NO:305); Lys Asp Pro Leu Phe Asn (SEQ ID NO:306); Leu Ser Gln His Thr Asn (SEQ ID NO:307); Asn Arg Leu Leu Leu Thr (SEQ ID NO:308); Tyr Pro Leu Trp Val Ile (SEQ ID NO:309); 20 Leu Leu Ile Ile Asp Arg (SEQ ID NO:310); Arg Val lie Ser Leu Gin (SEQ ID NO:311); Glu Val Ser Arg Glu Asp (SEQ ID NO:312); Ser Ile Leu Arg Ser Thr (SEQ ID NO:313); Pro Gly Leu Val Trp Leu (SEQ ID NO:314); 25 Val Lys Lys Leu Tyr Ile (SEQ ID NO:315); Asn Asn Arg Leu Leu Asp (SEQ ID NO:316); Ser Lys Gly Arg Trp Gly (SEQ ID NO:317); Ile Arg Pro Ser Gly Ile (SEQ ID NO:318); Ala Ser Leu Cys Pro Thr (SEQ ID NO:319); 30 Asp Val Pro Gly Leu Arg (SEQ ID NO:320); Arg His Arg Glu Val Gln (SEQ ID NO:321); Leu Ala Arg Lys Arg Ser (SEQ ID NO:322); 30 WO 2004/091493 PCT/US2004/010983 Ser Val Leu Asp His Val (SEQ ID NO:323); Asn Leu Leu Arg Arg Ala (SEQ ID NO:324); Ser Gly Ile Ser Ala Trp (SEQ ID NO:325); Phe Tyr Phe Trp Val Arg (SEQ 11D NO:326); 5 Lys Leu Phe Leu Pro Leu (SEQ ID NO:327); Thr Pro Thr Leu Ser Asp (SEQ ID NO:328); Thr His Ser Leu Ile Leu (SEQ ID NO:329); Leu Leu Leu Leu Ser Arg (SEQ ID NO:330); Leu Leu Arg Val Arg Ser (SEQ ID NO:331); 10 Glu Arg Arg ser Arg Gly (SEQ ID NO:332); Arg Met Leu Gln Leu Ala (SEQ ID NO:333); Age Gly Trp Ala Asn Ser (SEQ ID NO:334); Arg Pro Phe Tyr Ser Tyr (SEQ ID NO:335); Ser Ser Ser Trp Asn Ala (SEQ ID NO:336); 15 Leu Gly His Leu Glu Glu (SEQ ID NO:337); Ser Ala Val Thr Asn Thr (SEQ ID NO:338); Leu Arg Arg Ala Ser Leu (SEQ ID NO:339); Leu Arg Arg Trp Ser Leu (SEQ ID NO:340); Lys Trp Val His Leu Phe (SEQ ID NO:341); 20 Asn Arg Leu Leu Leu Thr (SEQ ID NO:342); Ala Arg Leu Leu Leu Thr (SEQ ID NO:343); Asn Ala Leu Leu Leu Thr (SEQ ID NO:344); Asn Arg Leu Ala Leu Thr (SEQ ID NO:345); Asn Leu Leu Arg Leu Thr (SEQ ID NO:346); 25 Asn Arg Leu Trp Leu Thr (SEQ ID NO:347); Asn Arg Leu Leu Leu Ala (SEQ ID NO:348); Met Gln Glu Arg Ile Thr Leu Lys Asp Tyr Ala Met (SEQ ID NO:349); Leu Arg Arg Trp Ser Leu Gly (SEQ ID NO:353); Lys Tirp Val His Leu Phe Gly (SEQ ID NO:354); 30 Asn Arg Leu Leu Leu Thr Gly (SEQ ID NO:355); Ala Arg Leu Leu Leu Thr Gly (SEQ ID NO:356); Asn Ala Leu Leu Leu Thr Gly (SEQ ID NO:357); 31 WO 2004/091493 PCT/US2004/010983 Asn Arg Leu Ala Leu Thr Gly (SEQ ID NO:358); Asn Leu Leu Arg Leu Thr Gly (SEQ ID NO:359); Asn Arg Leu Trp Leu Thr Gly (SEQ ID NO:360); Asn Arg Leu Leu Leu Ala Gly (SEQ ID NO:361); 5 Gly Lys Trp Val Tyr Ile Gly (SEQ ID NO:295); Ala Lys Arg Glu Thr Lys Gly (SEQ ID NO:296); Lys Trp Val His Leu Phe Gly (SEQ ID NO:297); Arg Leu Val Leu Val Leu Gly (SEQ ID NO:298); Trp Lys Trp Gly Ile Tyr (SEQ ID NO:299); 10 Set Ser His Ala Set Ala (SEQ ID NO:300); Trp Gly Pro Trp Ser Phe (SEQ ID NO:301); Ala Ile Pro Gly Lys Val (SEQ ID NO:302); Arg Val His Asp Pro Ala Gly (SEQ ID NO:303); Arg Ser Val Ser Ser Phe Gly (SEQ ID NO:304); 15 Leu Gly Thr Arg Lys Gly Gly (SEQ ID NO:305); Lys Asp Pro Leu Phe Asn Gly (SEQ ID NO:306); Leu Ser Gin His Thr Asn Gly (SEQ ID NO:307); Asn Arg Leu Leu Leu Thr Gly (SEQ ID NO:308); Tyr Pro Leu Trp Val Ile Gly (SEQ ID NO:309); 20 Leu Leu Ile Ile Asp Arg Gly (SEQ ID NO:310); Arg Val Ile Set Leu Gln Gly (SEQ ID NO:311); Glu Val Ser Arg Glu Asp Gly (SEQ ID NO:312); Ser Ile Leu Arg Ser Thr Gly (SEQ ID NO:313); Pro Gly Leu Val Trp Leu Gly (SEQ ID NO:314); 25 Val Lys Lys Leu Tyr Ile Gly (SEQ ID NO:315); Asn Asn Arg Leu Leu Asp Gly (SEQ ID NO:316); Ser Lys Gly Arg Trp Gly Gly (SEQ ID NO:317); Ile Arg Pro Ser Gly Ile Gly (SEQ ID NO:318); Ala Ser Leu Cys Pro Thr Gly (SEQ ID NO:319); 30 Asp Val Pro Gly Leu Arg Gly (SEQ ID NO:320); Arg His Arg Glu Val Gln Gly (SEQ ID NO:321); Leu Ala Arg Lys Arg Ser Gly (SEQ ID NO:322); 32 WO 2004/091493 PCT/US2004/010983 Ser Val Leu Asp His Val Gly (SEQ ID NO:323); Asn Leu Len Arg Arg Ala Gly (SEQ ID NO:324); Ser Gly Ile Ser Ala Trp Gly (SEQ ID NO:325); Phe Tyr Phe Trp Val Arg Gly (SEQ ID NO:326); 5 Lys Leu Phe Leu Pro Leu Gly (SEQ ID NO:327); Thr Pro Thr Leu Ser Asp Gly (SEQ ID NO:328); Thr His Ser Leu Ile Leu Gly (SEQ ID NO:329); Leu Leu Leu Leu Ser Arg Gly (SEQ ID NO:330); Leu Leu Arg Val Arg Ser Gly (SEQ ID NO:331); 10 Glu Arg Arg ser Arg Gly Gly (SEQ ID NO:332); Arg Met Leu Gln Leu Ala Gly (SEQ ID NO:333); Age Gly Trp Ala Asn Ser Gly (SEQ ID NO:334); Arg Pro Phe Tyr Ser Tyr Gly (SEQ ID NO:335); Ser Ser Ser Trp Asn Ala Gly (SEQ ID NO:336); 15 Leu Gly His Leu Glu Glu Gly (SEQ ID NO:337); and Ser Ala Val Thr Asn Thr Gly (SEQ ID NO:338); as described by Gragerov et al., J. Molec. Biol. 235:848-854 (1994). [0057] Other heat shock protein binding domains include Phe Tyr Gin Leu Ala Leu Thr(SEQ ID NO: ), Phe Tyr Gln Leu Ala Leu Thr Trp (SEQ ID NO: ), Arg Lys Leu Phe 20 Phe Asn Leu Arg (SEQ ID NO: ), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:), Lys Phe Glu Arg Gln (SEQ ID NO: ), Asn Ile Val Arg Lys Lys Lys (SEQ ID NO: ), and Arg Gly Tyr Val Tyr Gln Gly Leu (SEQ ID NO: ). [0058] Moreover, other heat shock protein binding domains include those described in WO9922761. Xaa represents any amino acid. 25 HTTVYGAG (SEQ ID NO: 82); TETPYPTG (SEQ ID NO: 83); LTTPFSSG (SEQ ID NO: 84); GVPLTMDG (SEQ ID NO: 85); KLPTVLRG (SEQ ID NO: 86); 30 CRFHGNRG (SEQ ID NO: 87); 33 WO 2004/091493 PCT/US2004/010983 YTRDFEAG (SEQ ID NO: 88); SSAAGPRG (SEQ ID NO: 89); SLIQYSRG (SEQ ID NO: 90); DALMWP Xaa G (SEQ ID NO: 91); 5 SS Xaa SLYIG (SEQ ID NO: 92); FNTSTRTG (SEQ ID NO: 93); TVQHVAFG (SEQ ID NO: 94); DYSFPPLG (SEQ ID NO: 95); VGSMESLG (SEQ ID NO: 96); 10 F Xaa PMI Xaa SG (SEQ ID NO: 97); APPRVTMG (SEQ ID NO: 98); IATKTPKG (SEQ ID NO: 99); KPPLFQIG (SEQ ID NO: 100); YHTAHNMG (SEQ ID NO: 101); 15 SYIQATHG (SEQ ID NO: 102); SSFATFLG (SEQ ID NO: 103); TTPPNFAG (SEQ ID NO: 104); ISLDPRMG (SEQ ID NO: 105); SLPLFGAG (SEQ ID NO: 106); 20 NLLKTTLG (SEQ ID NO: 107); DQNLPRRG (SEQ ID NO: 108); SHFEQLLG (SEQ ID NO: 109); TPQLHHGG (SEQ ID NO: 110); APLDRITG (SEQ ID NO: 111); 25 FAPLIAHG (SEQ ID NO: 112); SWIQTFMG (SEQ ID NO: 113); NTWPHMYG (SEQ ID NO: 114); EPLPTTLG (SEQ ID NO: 115); HGPHLFNG (SEQ ID NO: 116); 30 YLNSTLAG (SEQ ID NO: 117); HLHSPSGG (SEQ IDNO: 118); TLPHRLNG (SEQ ID NO: 119); 34 WO 2004/091493 PCT/US2004/010983 SSPREVHG (SEQ ID NO: 120); NQVDTARG (SEQ ID NO: 121); YPTPLLTG (SEQ ID NO: 122); HPAAFPWG (SEQ ID NO: 123); 5 LLPHSSAG (SEQ ID NO: 124); LETYTASG (SEQ ID NO: 125); KYVPLPPG (SEQ ID NO: 126); APLALHAG (SEQ ID NO: 127); YESLLTKG (SEQ ID NO: 128); 10 SHAASGTG (SEQ ID NO: 129); GLATVKSG (SEQ ID NO: 130); GATSFGLG (SEQ ID NO: 131); KPPGPVSG (SEQ ID NO: 132); TLYVSGNG (SEQ ID NO: 133); 15 HAPFKSQG (SEQ ID NO: 134); VAFTRLPG (SEQ ID NO: 135); LPTRTPAG (SEQ ID NO: 136); ASFDLLIG (SEQ ID NO: 137); RMNTEPPG (SEQ ID NO: 138); 20 KMTPLTTG (SEQ ID NO: 139); ANATPLLG (SEQ ID NO: 140); TIWPPPVG (SEQ ID NO: 141); QTKVMTTG (SEQ ID NO: 142); NHAVFASG (SEQ ID NO: 143); 25 LHAA Xaa TSG (SEQ ID NO: 144); TWQPYFHG (SEQ ID NO: 145); APLALHAG (SEQ ID NO: 146); TAHDLTVG (SEQ ID NO: 147); NMTNMLTG (SEQ ID NO: 148); 30 GSGLSQDG (SEQ ID NO: 149); TPIKTIYG (SEQ ID NO: 150); SHLYRSSG (SEQ ID NO: 151); 35 WO 2004/091493 PCT/US2004/010983 YTLVQPL (SEQ ID NO: 152); TPDITPK (SEQ ID NO: 153); TYPDLRY (SEQ ID NO: 154); DRTHATS (SEQ ID NO: 155); 5 MSTTFYS (SEQ ID NO: 156); YQHAVQT (SEQ ID NO: 157); FPFSAST (SEQ ID NO: 158); SSFPPLD (SEQ ID NO: 159); MAPSPPH (SEQ ID NO: 160); 10 SSFPDLL (SEQ ID NO: 161); HSYNRLP (SEQ ID NO: 162); HLTHSQR (SEQ IDNO: 163); QAAQSRS (SEQ ID NO: 164); FATHHIG (SEQ ID NO: 165); 15 SMPEPLI (SEQ ID NO: 166); IPRYHLI (SEQ ID NO: 167); SAPHMTS (SEQ ID NO: 168); KAPVWAS (SEQ ID NO: 169); LPHWLLI (SEQ ID NO: 170); 20 ASAGYQI (SEQ ID NO: 171); VTPKTGS (SEQ ID NO: 172); EHPMPVL (SEQ ID NO: 173); VSSFVTS (SEQ ID NO: 174); STHFTWP (SEQ ID NO: 175); 25 GQWWSPD (SEQ ID NO: 176); GPPHQDS (SEQ ID NO: 177); NTLPSTI (SEQ ID NO: 178); HQPSRWV (SEQ ID NO: 179); YGNPLQP (SEQ ID NO: 180); 30 FHWWWQP (SEQ ID NO: 181); ITLKYPL (SEQ ID NO: 182); FHIWPWLF (SEQ ID NO: 183); 36 WO 2004/091493 PCT/US2004/010983 TAQDSTG (SEQ ID NO: 184); FHWWWQP (SEQ ID NO: 185); FHWWDWW (SEQ ID NO: 186); EPFFRMQ (SEQ ID NO: 187); 5 TWWLNYR (SEQ ID NO: 188); FHWWWQP (SEQ ID NO: 189); QPSHLRW (SEQ ID NO: 190); SPASPVY (SEQ ID NO: 191); FHWWWQP (SEQ ID NO: 192); 10 HPSNQAS (SEQ ID NO: 193); NSAPRPV (SEQ ID NO: 194); QLWSIYP (SEQ ID NO: 195); SWPFFDL (SEQ ID NO: 196); DTTLPLH (SEQ ID NO: 197); 15 WHWQMLW (SEQ ID NO: 198); DSFRTPV (SEQ ID NO: 199); TSPLSLL (SEQ ID NO: 200); AYNYVSD (SEQ ID NO: 201); RPLHDPM (SEQ ID NO: 202); 20 WPSTTLF (SEQ ID NO: 203); ATLEPVR (SEQ ID NO: 204); SMTVLRP (SEQ ID NO: 205); QIGAPSW (SEQ ID NO: 206); APDLYVP (SEQ ID NO: 207); 25 RMPPLLP (SEQ ID NO: 208); AKATPEH (SEQ ID NO: 209); TPPLRIN (SEQ ID NO: 210); LPIHAPH (SEQ ID NO: 211); DLNAYTH (SEQ ID NO: 212); 30 VTLPNFH (SEQ ID NO: 213); NSRLPTL (SEQ ID NO: 214); YPHPSRS (SEQ ID NO: 215); 37 WO 2004/091493 PCT/US2004/010983 GTAHFMY (SEQ ID NO: 216); YSLLPTR (SEQ ID NO: 217); LPRRTLL (SEQ ID NO: 218); TSTLLWK (SEQ ID NO: 219); 5 TSDMKPH (SEQ ID NO: 220); TSSYLAL (SEQ ID NO: 221); NLYGPHD (SEQ ID NO: 222); LETYTAS (SEQ ID NO: 223); AYKSLTQ (SEQ ID NO: 224); 10 STSVYSS (SEQ ID NO: 225); EGPLRSP (SEQ ID NO: 226); TTYHALG (SEQ ID NO: 227); VSIGI-IPS (SEQ ID NO: 228); THSHRPS (SEQ ID NO: 229); 15 ITNPLTT (SEQ ID NO: 230); SIQAHHS (SEQ IDNO:231); LNWPRVL (SEQ ID NO:232); YYYAPPP (SEQ ID NO:233); SLWTRLP (SEQ ID NO:234); 20 NVYHSSL (SEQ ID NO:235); NSPHPPT (SEQ ID NO:236); VPAKPRH (SEQ ID NO:237); HNLHPNR (SEQ ID NO:238); YTTHRWL (SEQ ID NO:239); 25 AVTAAIV (SEQ ID NO:240); TLMHDRV (SEQ ID NO:241); TPLKVPY (SEQ ID NO:242); FTNQQYH (SEQ ID NO:243); SHVPSMA (SEQ ID NO:244); 30 HTTVYGA (SEQ ID NO:245); TETPYPT (SEQ ID NO:246); LTTPFSS (SEQ ID NO:247); 38 WO 2004/091493 PCT/US2004/010983 GVPLTMD (SEQ ID NO:248); KLPTVLR (SEQ ID NO:249); CRFHGNR (SEQ ID NO:250); YTRDFEA (SEQ ID NO: 251); 5 SSAAGPR (SEQ ID NO: 252); SLIQYSR (SEQ ID NO: 253); DALMWP Xaa (SEQ ID NO: 254); SS Xaa SLYI (SEQ ID NO: 255); FNTSTRT (SEQ ID NO: 256); 10 TVQIIVAF (SEQ ID NO: 257); DYSFPPL (SEQ ID NO: 258); VGSMESL (SEQ ID NO: 259); F Xaa PMI Xaa S (SEQ ID NO: 260); APPRVTM (SEQ ID NO: 261); 15 IATKTPK (SEQ ID NO: 262); KPPLFQI (SEQ ID NO: 263); YHTAHNM (SEQ ID NO: 264); SYIQATH (SEQ ID NO: 265); SSFATFL (SEQ ID NO: 266); 20 TTPPNFA (SEQ ID NO: 267); ISLDPRM (SEQ ID NO: 268); SLPLFGA (SEQ ID NO: 269); NLLKTTL (SEQ ID NO: 270); DQNLPRR (SEQ ID NO: 271); 25 SHFEQLL (SEQ ID NO: 272); TPQLHHG (SEQ ID NO: 273); APLDRIT (SEQ ID NO: 274); FAPLIAH (SEQ ID NO: 275); SWTQTFM (SEQ ID NO: 276); 30 NTWPHMY (SEQ ID NO: 277); EPLPTTL (SEQ ID NO: 278); HGPHLFN (SEQ ID NO: 279); 39 WO 2004/091493 PCT/US2004/010983 YLNSTLA (SEQ ID NO: 280); HLHSPSG (SEQ ID NO: 281); TLPHRLN (SEQ ID NO: 282); SSPREVH (SEQ ID NO: 283); 5 NQVDTAR (SEQ ID NO: 284); YPTPLLT (SEQ ID NO: 285); HPAAFPW (SEQ ID NO: 286); LLPHSSA (SEQ ID NO: 287); LETYTAS (SEQ ID NO: 288); 10 KYVPLPP (SEQ ID NO: 289); APLALHA (SEQ ID NO: 290); YESLLTK (SEQ ID NO: 291); SHAASGT (SEQ ID NO: 292); GLATVKS (SEQ ID NO: 293); 15 GATSFGL (SEQ ID NO: 294); KPPGPVS (SEQ ID NO: 295); TLYVSGN (SEQ ID NO: 296); HAPFKSQ (SEQ ID NO: 297); VAFTRLP (SEQ ID NO: 298); 20 LPTRTPA (SEQ ID NO: 299); ASFDLLI (SEQ ID NO: 300); RMNTEPP (SEQ ID NO: 301); KMTPLTT (SEQ ID NO: 302); ANATPLL (SEQ ID NO: 303); 25 TIWPPPV (SEQ ID NO: 304); QTKVMTT (SEQ ID NO: 305); NHAVFAS (SEQ ID NO: 306); LHAA Xaa TS (SEQ ID NO: 307); TWQPYFH (SEQ ID NO: 308); 30 APLALHA (SEQ ID NO: 309); TAHDLTV (SEQ ID NO: 310); NMTNMLT (SEQ ID NO: 311); 40 WO 2004/091493 PCT/US2004/010983 GSGLSQD (SEQ ID NO: 312); TPIKTIY (SEQ ID NO: 313); SHLYRSS (SEQ ID NO: 314); HGQAWQF (SEQ ID NO: 315); and 5 FHWWW (SEQ ID NO: 317). [0059] The aforementioned heat shock protein binding domains are merely exemplary of various peptides, among peptide and non-peptide heat shock protein binding molecules, that may be used in the practice of the present invention. In other embodiments, the heat shock protein binding domain may be directed to bind to a different part of the mammalian heat 10 shock protein that those aforementioned, and the heat shock protein-binding domains of the invention are not limited to binding to any particular portion of the heat shock protein molecule. In a non-limiting example, the peptide IFAGIKKKAERADLIAYLKQATAK (Greene et al., 1995, J. Biol. Chem. 270:2967-2973; SEQ ID NO:331) or a heat shock protein-binding fragment of this peptide, is used in any of the conjugates of the invention to 15 facilitate the binding of a pre-selected molecule to a heat shock protein. In addition to the aforementioned peptides that bind to heat shock proteins, the binding may be achieved through the use of an organic molecule or compound with heat shock protein binding activity. For example, suitable molecules include members of the benzoquinone ansamycin antibiotics, such as herbimycin A, geldanamycin, macmimycin I, mimosamycin, and 20 kuwaitimycin (Omura et al., 1979, J. Antibiotics 32:255-261; see also WO9922761, incorporated by reference herein in its entirety), or structurally related compounds, and analogs or derivatives thereof. These molecules may be conjugated though established chemical means to the antigenic domains of the invention, via the peptide linker, to produce hybrid antigens capable of binding to a heat shock protein in vitro or in vivo and eliciting an 25 immune response to the antigen present therein. [0060] As described in co-pending and commonly-owned application serial no. 10/776,521, filed February 12, 2004, incorporated herein by reference in its entirety, it has been found that incorporation of a tryptophan residue (Trp, or single amino acid code W) at the C-terminus of the heat shock protein binding domains such as but not limited to those 30 identified as described above, enhances binding to heat shock proteins. Increased binding to heat shock proteins has been found to increase the ability of hybrid antigens to induce an 41 WO 2004/091493 PCT/US2004/010983 immune response to the antigenic domain of the hybrid antigen, whether administered in a complex with a heat shock protein or when administered alone. Increased immune response is correlated with increased efficacy of treating disease.Other examples of methods for determining affinity are described in PCT/US96/13363 (WO9706821), which is 5 incorporated herein by reference in its entirety. [0061] Among the foregoing selection of heat shock protein binding domains, those preferred in the present invention as part of a hybrid antigen comprising an antigenic domain and peptide linker of the invention there between includes the following heat shock protein binding domains: 10 [0062] Gly Lys Trp Val Tyr Ile Gly Trp (SEQ ID NO:); Ala Lys Arg Glu Thr Lys Gly Trp (SEQ ID NO:); Lys Trp Val His Leu Phe Gly Trp (SEQ ID NO:); Arg Leu Val Leu Val Leu Gly Trp (SEQ ID NO:); Trp Lys Trp Gly Ile Tyr Gly Trp (SEQ ID NO:); 15 Ser Ser His Ala Ser Ala Gly Trp (SEQ ID NO:); Trp Gly Pro Trp Ser Phe Gly Trp (SEQ ID NO:); Ala Ile Pro Gly Lys Val Gly Trp (SEQ ID NO:); Arg Val His Asp Pro Ala Gly Trp (SEQ ID NO:); Arg Ser Val Ser Ser Phe Gly Trp (SEQ ID NO:); 20 Leu Gly Thr Arg Lys Gly Gly Trp (SEQ ID NO:); Lys Asp Pro Leu Phe Asn Gly Trp (SEQ ID NO:); Leu Ser Gln His Thr Asn Gly Trp (SEQ ID NO:); Asn Arg Leu Leu Leu Thr Gly Trp (SEQ ID NO:); Tyr Pro Leu Trp Val Ile Gly Trp (SEQ ID NO:); 25 Leu Leu lie Ile Asp Arg Gly Trp (SEQ ID NO:); Arg Val Ile Ser Leu Gin Gly Trp (SEQ ID NO:); Glu Val Ser Arg Glu Asp Gly Trp (SEQ ID NO:); Ser Ile Leu Arg Ser Thr Gly Trp (SEQ ID NO:); Pro Gly Leu Val Trp Leu Gly Trp (SEQ ID NO:); 30 Val Lys Lys Leu Tyr Ile Gly Trp (SEQ ID NO:); Asn Asn Arg Leu Leu Asp Gly Trp (SEQ ID NO:); 42 WO 2004/091493 PCT/US2004/010983 Ser Lys Gly Arg Trp Gly Gly Trp (SEQ ID NO:); Ile Arg Pro Ser Gly Ile Gly Trp (SEQ ID NO:); Ala Ser Leu Cys Pro Thr Gly Trp (SEQ ID NO:); Asp Val Pro Gly Leu Arg Gly Trp (SEQ ID NO:); 5 Arg His Arg Glu Val Gin Gly Trp (SEQ ID NO:); Leu Ala Arg Lys Arg Ser Gly Trp (SEQ ID NO:); Ser Val Leu Asp His Val Gly Trp (SEQ ID NO:); Asn Leu Leu Arg Arg Ala Gly Trp (SEQ ID NO:); Set Gly Ile Ser Ala Trp Gly Trp (SEQ ID NO:); 10 Phe Tyr Phe Trp Val Arg Gly Trp (SEQ ID NO:); Lys Leu Phe Leu Pro Leu Gly Trp (SEQ ID NO:); Thr Pro Thr Leu Ser Asp Gly Trp (SEQ ID NO:); Thr His Ser Leu Ile Leu Gly Trp (SEQ ID NO:); Leu Leu Leu Leu Ser Arg Gly Trp (SEQ ID NO:); 15 Leu Leu Arg Val Arg Scr Gly Trp (SEQ ID NO:); Glu Arg Arg set Arg Gly Gly Trp (SEQ ID NO:); Arg Met Leu Gin Leu Ala Gly Trp (SEQ ID NO:); Age Gly Trp Ala Asn Ser Gly Trp (SEQ ID NO:); Arg Pro Phe Tyr Ser Tyr Gly Trp (SEQ ID NO:); 20 Set Ser Ser Trp Asn Ala Gly Trp (SEQ ID NO:); Leu Gly His Leu Glu Glu Gly Tip (SEQ ID NO:); Ser Ala Val Thr Asn Thr Gly Trp (SEQ ID NO:); Phe Tyr Gin Leu Ala Leu Thr (SEQ ID NO:); Phe Tyr Gln Leu Ala Leu Thr Trp (SEQ ID NO:), 25 Arg Lys Leu Phe Phe Asn Leu Arg (SEQ ID NO:), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:), Lys Phe Glu Arg Gin (SEQ ID NO: ), Asn Ile Val Arg Lys Lys Lys (SEQ ID NO: ), and Arg Gly Tyr Val Tyr Gln Gly Leu (SEQ ID NO:). 30 [0063] Other non-limiting examples of such heat shock protein binding domains with a terminal Trp residue useful for the various aspects of the present invention include: 43 WO 2004/091493 PCT/US2004/010983 [0064] Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:350); Phe Tyr Gin Leu Ala Leu Tyr Trp (SEQ ID NO:35 1); Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:352); Gly Lys Trp Val Tyr Ile Gly Trp (SEQ ID NO:295); 5 Ala Lys Arg Glu Thr Lys Gly Trp (SEQ ID NO:296); Lys Trp Val His Leu Phe Gly Trp (SEQ ID NO:297); Arg Leu Val Leu Val Leu Gly Trp (SEQ ID NO:298); Trp Lys Trp Gly Ile Tyr Gly Trp (SEQ ID NO:299); Ser Ser His Ala Ser Ala Gly Trp (SEQ ID NO:300); 10 Trp Gly Pro Trp Ser Phe Gly Trp (SEQ ID NO:301); Ala Ile Pro Gly Lys Val Gly Trp (SEQ ID NO:302); Arg Val His Asp Pro Ala Gly Trp (SEQ ID NO:303); Arg Ser Val Ser Ser Phe Gly Trp (SEQ ID NO:304); Leu Gly Thr Arg Lys Gly Gly Trp (SEQ ID NO:305); 15 Lys Asp Pro Leu Phe Asn Gly Trp (SEQ ID NO:306); Leu Ser Gln His Thr Asn Gly Tip (SEQ ID NO:307); Asn Arg Leu Leu Leu Thr Gly Tp (SEQ ID NO:308); Tyr Pro Leu Trp Val Ile Gly Tp (SEQ ID NO:309); Leu Leu Ile Ile Asp Arg Gly Trp (SEQ ID NO:310); 20 Arg Val Ile Ser Leu Gln Gly Trp (SEQ ID NO:311); Glu Val Ser Arg Glu Asp Gly Tp (SEQ ID NO:312); Ser lie Leu Arg Ser Thr Gly Tip (SEQ ID NO:313); Pro Gly Leu Val Tp Leu Gly Tp (SEQ ID NO:314); Val Lys Lys Leu Tyr Ile Gly Tp (SEQ ID NO:315); 25 Asn Asn Arg Leu Leu Asp Gly Tp (SEQ ID NO:316); Ser Lys Gly Arg Tp Gly Gly Tp (SEQ ID NO:317); Ile Arg Pro Ser Gly Ile Gly Tp (SEQ ID NO:318); Ala Ser Leu Cys Pro Thr Gly Tp (SEQ ID NO:319); Asp Val Pro Gly Leu Arg Gly Tp (SEQ ID NO:320); 30 Arg His Arg Glu Val Gin Gly Tp (SEQ ID NO:321); Len Ala Arg Lys Arg Ser Gly Tip (SEQ ID NO:322); Ser Val Leu Asp His Val Gly Trp (SEQ ID NO:323); 44 WO 2004/091493 PCT/US2004/010983 Asn Leu Leu Arg Arg Ala Gly Trp (SEQ ID NO:324); Ser Gly Ile Ser Ala Trp Gly Trp (SEQ ID NO:325); Phe Tyr Phe Trp Val Arg Gly Tp (SEQ ID NO:326); Lys Leu Phe Leu Pro Len Gly Tp (SEQ ID NO:327); 5 Thr Pro Thr Leu Ser Asp Gly Trp (SEQ ID NO:328); Thr His Ser Leu Ile Leu Gly Tp (SEQ ID NO:329); Leu Leu Leu Leu Ser Arg Gly Trp (SEQ ID NO:330); Leu Leu Arg Val Arg Ser Gly Trp (SEQ ID NO:331); Glu Arg Arg ser Arg Gly Gly Trp (SEQ ID NO:332); 10 Arg Met Leu Gin Leu Ala Gly Tp (SEQ ID NO:333); Age Gly Trp Ala Asn Ser Gly Trp (SEQ ID NO:334); Arg Pro Phe Tyr Ser Tyr Gly Tp (SEQ ID NO:335); Ser Ser Ser Trp Asn Ala Gly Trp (SEQ ID NO:336); Leu Gly His Leu Glu Glu Gly Trp (SEQ ID NO:337); 15 Ser Ala Val Thr Asn Thr Gly Trp (SEQ ID NO:338); Leu Arg Arg Ala Ser Leu Trp (SEQ ID NO:339); Leu Arg Arg Trp Ser Leu Trp (SEQ ID NO:340); Lys Trp Val His Leu Phe Trp (SEQ ID NO:341); Asn Arg Leu Leu Leu Thr Tp (SEQ ID NO:342); 20 Ala Arg Leu Leu Leu Thr Tp (SEQ ID NO:343); Asn Ala Leu Leu Leu Thr Tp (SEQ ID NO:344); Asn Arg Leu Ala Leu Thr Tp (SEQ ID NO:345); Asn Leu Leu Arg Leu Thr Tip (SEQ ID NO:346); Asn Arg Leu Trp Leu Thr Tip (SEQ ID NO:347); and 25 Asn Arg Leu Leu Leu Ala Trp (SEQ ID NO:348). [0065] Other heat shock protein binding domains useful in the practice of the present invention include Phe Tyr Gln Len Ala Leu Thr Trp (SEQ ID NO:501), Phe Tyr Gin Leu Ala Leu Thr Tip (SEQ ID NO:502), Arg Lys Leu Phe Phe Asn Leu Arg Tp (SEQ ID NO:503 ), Arg Lys Leu Phe Phe Asn Leu Arg Tip (SEQ ID NO:504 ), Lys Phe 30 Glu Arg Gin Tp (SEQ ID NO:505), Asn Ile Val Arg Lys Lys Lys Tp (SEQ ID NO:506), and Arg Gly Tyr Val Tyr Gin Gly Leu Trp (SEQ ID NO:507). 45 WO 2004/091493 PCT/US2004/010983 [0066] Moreover, other heat shock protein binding domains include those described in WO9922761, and may have a terminal Trp residue added to achieve the purposes of the present invention. Xaa represents any amino acid. Tyr Thr Leu Val Gin Pro Leu Trp (SEQ ID NO: 1149); 5 Thr Pro Asp Ile Thr Pro Lys Trp (SEQ ID NO: 1150); Thr Tyr Pro Asp Leu Arg Tyr Trp (SEQ ID NO: 1151); Asp Arg Thr His Ala Thr Ser Trp (SEQ ID NO: 1152); Met Ser Thr Thr Phe Tyr Ser Trp (SEQ ID NO: 1153); Tyr Gin His Ala Val Gln Thr Trp (SEQ ID NO: 1154); 10 Phe Pro Phe Ser Ala Ser Thr Trp (SEQ ID NO: 1155); Ser Ser Phe Pro Pro Leu Asp Trp (SEQ ID NO: 1156); Met Ala Pro Ser Pro Pro His Trp (SEQ ID NO: 1157); Ser Ser Phe Pro Asp Leu Leu Trp (SEQ ID NO: 1158); His Ser Tyr Asn Arg Leu Pro Trp (SEQ ID NO: 1159); 15 His Leu Thr His Ser Gln Arg Trp (SEQ ID NO: 1160); Gin Ala Ala Gln Ser Arg Ser Trp (SEQ ID NO: 1161); Phe Ala Thr His His Ile Gly Trp (SEQ ID NO: 1162); Ser Met Pro Glu Pro Leu Ile Trp (SEQ ID NO: 1163); lie Pro Arg Tyr His Leu Ile Trp (SEQ ID NO: 1164); 20 Ser Ala Pro His Met Thr Ser Trp (SEQ ID NO: 1165); Lys Ala Pro Val Trp Ala Ser Trp (SEQ ID NO: 1166); Leu Pro His Trp Leu Leu Ile Trp (SEQ ID NO: 1167); Ala Ser Ala Gly Tyr Gln Ile Trp (SEQ ID NO: 1168); Val Thr Pro Lys Thr Gly Ser Trp (SEQ ID NO: 1169); 25 Glu His Pro Met Pro Val Leu Trp (SEQ ID NO: 1170); Val Ser Ser Phe Val Thr Ser Trp (SEQ ID NO: 1171); Ser Thr His Phe Thr Trp Pro Trp (SEQ ID NO: 1172); Gly Gin Trp Tip Ser Pro Asp Trp (SEQ ID NO: 1173); Gly Pro Pro His Gln Asp Ser Trp (SEQ ID NO: 1174); 30 Asn Thr Leu Pro Ser Thr Ile Trp (SEQ ID NO: 1175); His Gln Pro Ser Arg Tip Val Trp (SEQ ID NO: 1176); Tyr Gly Asn Pro Leu Gn Pro Trp (SEQ ID NO: 1177); 46 WO 2004/091493 PCT/US2004/010983 Phe His Trp Trp Trp Gin Pro Trp (SEQ ID NO: 1178); Ile Thr Leu Lys Tyr Pro Leu Trp (SEQ ID NO: 1179); Phe His Tip Pro Tp Leu Phe Tp (SEQ ID NO: 1180); Thr Ala Gin Asp Ser Thr Gly Trp (SEQ ID NO: 1181); 5 Phe His Trp Trp Trp Gln Pro Trp (SEQ ID NO: 1182); Phe His Trp Tip Asp Tp Tp Trp (SEQ ID NO: 1183); Glu Pro Phe Phe Arg Met Gin Tp (SEQ ID NO: 1184); Thr Trp Tp Leu Asn Tyr Arg Tp (SEQ ID NO: 1185); Phe His Trp Trp Tp Gin Pro Tip (SEQ ID NO: 1186); 10 Gin Pro Ser His Leu Arg Trp Trp (SEQ ID NO: 1187); Ser Pro Ala Ser Pro Val Tyr Trp (SEQ ID NO: 1188); Phe His Tp Tp Tp Gin Pro Trp (SEQ ID NO: 1189); His Pro Ser Asn Gin Ala Ser Tp (SEQ ID NO: 1190); Asn Ser Ala Pro Arg Pro Val Tip (SEQ ID NO: 1191); 15 Gln Leu Trp Ser Ile Tyr Pro Trp (SEQ ID NO: 1192); Ser Trp Pro Phe Phe Asp Leu Tip (SEQ ID NO: 1193); Asp Thr Thr Leu Pro Leu His Tp (SEQ ID NO: 1194); Tip His Tp Gin Met Leu Tp Tp (SEQ ID NO: 1195); Asp Ser Phe Arg Thr Pro Val Trp (SEQ ID NO: 1196); 20 Thr Ser Pro Len Ser Leu Leu Tp (SEQ ID NO: 1197); Ala Tyr Asn Tyr Val Ser Asp Tp (SEQ ID NO: 1198); Arg Pro Leu His Asp Pro Met Tp (SEQ ID NO: 1199); Tip Pro Ser Thr Thr Leu Phe Tp (SEQ ID NO: 1200); Ala Thr Leu Glu Pro Val Arg Tip (SEQ ID NO: 1201); 25 Ser Met Thr Val Leu Arg Pro Tp (SEQ ID NO: 1202); Gin Ile Gly Ala Pro Ser Tip Tp (SEQ ID NO: 1203); Ala Pro Asp Leu Tyr Val Pro Trp (SEQ ID NO: 1204); Arg Met Pro Pro Leu Leu Pro Tp (SEQ ID NO: 1205); Ala Lys Ala Thr Pro Glu His Trp (SEQ ID NO: 1206); 30 Thr Pro Pro Leu Arg Ile Asn Tp (SEQ ID NO: 1207); Leu Pro Ile His Ala Pro His Tp (SEQ ID NO: 1208); Asp Leu Asn Ala Tyr Thr His Trp (SEQ ID NO: 1209); 47 WO 2004/091493 PCT/US2004/010983 Val Thr Leu Pro Asn Phe His Trp (SEQ ID NO: 1210); Asn Set Arg Leu Pro Thr Leu Tip (SEQ ID NO: 1211); Tyr Pro His Pro Ser Arg Ser Trp (SEQ ID NO: 1212); Gly Thr Ala His Phe Met Tyr Trp (SEQ ID NO: 1213); 5 Tyr Ser Leu Leu Pro Thr Arg Trp (SEQ ID NO: 1214); Leu Pro Arg Arg Thr Leu Leu Trp (SEQ ID NO: 1215); Thr Ser Thr Leu Leu Trp Lys Trp (SEQ ID NO: 1216); Thr Ser Asp Met Lys Pro His Trp (SEQ ID NO: 1217); Thr Set Ser Tyr Leu Ala Leu Trp (SEQ ID NO: 1218); 10 Asn Leu Tyr Gly Pro His Asp Trp (SEQ ID NO: 1219); Leu Glu Thr Tyr Thr Ala Ser Trp (SEQ ID NO: 1220); Ala Tyr Lys Ser Leu Thr Gin Trp (SEQ ID NO: 1221); Set Thr Ser Val Tyr Set Ser Tirp (SEQ ID NO: 1222); Glu Gly Pro Leu Arg Set Pro Trp (SEQ ID NO: 1223); 15 Thr Thr Tyr His Ala Leu Gly Trp (SEQ ID NO: 1224); Val Set Ile Gly His Pro Set Tip (SEQ ID NO: 1225); Thr His Ser His Arg Pro Ser Trp (SEQ ID NO: 1226); Ile Thr Asn Pro Leu Thr Thr Tip (SEQ ID NO: 1227); Ser Ile Gln Ala His His Ser Trp (SEQ ID NO: 1228); 20 Leu Asn Trp Pro Arg Val Leu Trp (SEQ ID NO: 1229); Tyr Tyr Tyr Ala Pro Pro Pro Trp (SEQ ID NO: 1230); Set Leu Trp Thr Arg Leu Pro Trp (SEQ ID NO: 1231); Asn Val Tyr His Ser Ser Leu Trp (SEQ ID NO: 1232); Asn Ser Pro His Pro Pro Thr Trp (SEQ ID NO: 1233); 25 Val Pro Ala Lys Pro Arg His Trp (SEQ ID NO: 1234); His Asn Leu His Pro Asn Arg Trp (SEQ ID NO: 1235); Tyr Thr Thr His Arg Trp Leu Trp (SEQ ID NO: 1236); Ala Val Thr Ala Ala Ile Val Tip (SEQ ID NO: 1237); Thr Leu Met His Asp Arg Val Trp (SEQ ID NO: 1238); 30 Thr Pro Leu Lys Val Pro Tyr Trp (SEQ ID NO: 1239); Phe Thr Asn Gln Gln Tyr His Trp (SEQ ID NO: 1240); Ser His Val Pro Ser Met Ala Tip (SEQ ID NO: 1241); 48 WO 2004/091493 PCT/US2004/010983 His Thr Thr Val Tyr Gly Ala Trp (SEQ ID NO: 1242); Thr Glu Thr Pro Tyr Pro Thr Trp ( SEQ ID NO: 1243); Leu Thr Thr Pro Phe Ser Ser Trp (SEQ ID NO: 1244); Gly Val Pro Leu Thr Met Asp Trp (SEQ ID NO: 1245); 5 Lys Leu Pro Thr Val Leu Arg Trp (SEQ ID NO: 1246); Cys Arg Phe His Gly Asn Arg Trp (SEQ ID NO: 1247); Tyr Thr Arg Asp Phe Glu Ala Trp (SEQ ID NO: 1248); Ser Ser Ala Ala Gly Pro Arg Trp (SEQ ID NO: 1249); Ser Leu Ile Gin Tyr Ser Arg Trp (SEQ ID NO: 1250); 10 Asp Ala Leu Met Trp Pro XAA Trp (SEQ ID NO: 1251); Ser Ser XAA Ser Leu Tyr Ile Trp (SEQ ID NO: 1252); Phe Asn Thr Ser Thr Arg Thr Trp (SEQ ID NO: 1253); Thr Val Gin His Val Ala Phe Trp (SEQ ID NO: 1254); Asp Tyr Set Phe Pro Pro Leu Trp (SEQ ID NO: 1255); 15 Val Gly Ser Met Glu Ser Leu Trp (SEQ ID NO: 1256); Phe XAA Pro Met Ile XAA Ser Trp (SEQ ID NO: 1257); Ala Pro Pro Arg Val Thr Met Trp (SEQ ID NO: 1258); Ile Ala Thr Lys Thr Pro Lys Trp (SEQ ID NO: 1259); Lys Pro Pro Leu Phe Gin Ile Trp (SEQ ID NO: 1260); 20 Tyr His Thr Ala His Asn Met Trp (SEQ ID NO: 1261); Ser Tyr lie Gin Ala Thr His Trp (SEQ ID NO: 1262); Ser Ser Phe Ala Thr Phe Leu Trp (SEQ ID NO: 1263); Thr Thr Pro Pro Asn Phe Ala Tip (SEQ ID NO: 1264); Ile Ser Leu Asp Pro Arg Met Trp (SEQ ID NO: 1265); 25 Ser Leu Pro Leu Phe Gly Ala Trp (SEQ ID NO: 1266); Asn Leu Leu Lys Thr Thr Leu Trp (SEQ ID NO: 1267); Asp Gln Asn Leu Pro Arg Arg Trp (SEQ ID NO: 1268); Ser His Phe Glu Gin Leu Leu Trp (SEQ ID NO: 1269); Thr Pro Gin Leu His His Gly Trp (SEQ ID NO: 1270); 30 Ala Pro Leu Asp Arg Ile Thr Trp (SEQ ID NO: 1271); Phe Ala Pro Leu Ile Ala His Trp (SEQ ID NO: 1272); Ser Trp Ile Gin Thr Phe Met Trp (SEQ ID NO: 1273); 49 WO 2004/091493 PCT/US2004/010983 Asn Thr Trp Pro His Met Tyr Trp (SEQ ID NO: 1274); Glu Pro Leu Pro Thr Thr Leu Trp (SEQ ID NO: 1275); His Gly Pro His Leu Phe Asn Trp (SEQ ID NO: 1276); Tyr Leu Asn Ser Thr Leu Ala Trp (SEQ ID NO: 1277); 5 His Leu His Ser Pro Ser Gly Trp (SEQ ID NO: 1278); Thr Leu Pro His Arg Leu Asn Trp (SEQ ID NO: 1279); Set Ser Pro Arg Glu Val His Tip (SEQ ID NO: 1280); Asn Gln Val Asp Thr Ala Arg Tp (SEQ ID NO: 1281); Tyr Pro Thr Pro Leu Leu Thr Tip (SEQ ID NO: 1282); 10 His Pro Ala Ala Phe Pro Trp Tp (SEQ ID NO: 1283); Leu Leu Pro His Ser Ser Ala Tp (SEQ ID NO: 1284); Leu Glu Thr Tyr Thr Ala Ser Tp (SEQ ID NO: 1285); Lys Tyr Val Pro Leu Pro Pro Tip (SEQ ID NO: 1286); Ala Pro Leu Ala Leu His Ala Tp (SEQ ID NO: 1287); 15 Tyr Glu Ser Leu Leu Thr Lys Tp (SEQ ID NO: 1288); Ser His Ala Ala Ser Gly Thr Tp (SEQ ID NO: 1289); Gly Leu Ala Thr Val Lys Set Tp (SEQ ID NO: 1290); Gly Ala Thr Ser Phe Gly Leu Tp (SEQ ID NO: 1291); Lys Pro Pro Gly Pro Val Set Tp (SEQ ID NO: 1292); 20 Thr Leu Tyr Val Ser Gly Asn Tp (SEQ ID NO: 1293); His Ala Pro Phe Lys Ser Gln Tp (SEQ ID NO: 1294); Val Ala Phe Thr Arg Leu Pro Tp (SEQ ID NO: 1295); Leu Pro Thr Arg Thr Pro Ala Tp (SEQ ID NO: 1296); Ala Set Phe Asp Leu Leu Ile Tip (SEQ ID NO: 1297); 25 Arg Met Asn Thr Glu Pro Pro Tip (SEQ ID NO: 1298); Lys Met Thr Pro Leu Thr Thr Trp (SEQ ID NO: 1299); Ala Asn Ala Thr Pro Leu Leu Tp (SEQ ID NO: 1300); Thr Ile Tp Pro Pro Pro Val Tp (SEQ ID NO: 1301); Gln Thr Lys Val Met Thr Thr Trp (SEQ ID NO: 1302); 30 Asn His Ala Val Phe Ala Set Tp (SEQ ID NO: 1303); Leu His Ala Ala Xaa Thr Ser Tp (SEQ ID NO: 1304); Thr Tp Gln Pro Tyr Phe His Tp (SEQ ID NO: 1305); 50 WO 2004/091493 PCT/US2004/010983 Ala Pro Leu Ala Leu His Ala Trp (SEQ ID NO: 1306); Thr Ala His Asp Leu Thr Val Tip (SEQ ID NO: 1307); Asn Met Thr Asn Met Leu Thr Trp (SEQ ID NO: 1308); Gly Ser Gly Leu Ser Gn Asp Tip (SEQ ID NO: 1309); 5 Thr Pro le Lys Thr lle Tyr Trp (SEQ ID NO: 1310); Ser His Leu Tyr Arg Ser Set Trp (SEQ ID NO: 1311); and His Gly Gin Ala Trp Gln Phe Trp (SEQ ID NO: 1312). [0067] Among all of the foregoing heat shock protein binding peptides, the heat shock protein binding domain Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:350) is most 10 preferred in the hybrid antigens of the invention. However, the aforementioned heat shock protein binding domains are merely exemplary of various moieties, among peptide and non peptide heat shock protein binding molecules, that may be used in the practice of the present invention. [0068] The hybrid antigen of the invention incorporates at least one antigenic 15 (immunogenic) domain and at least one one heat shock protein-binding domain, separated by at least one peptide linker as described herein. The hybrid antigen of the invention may be synthesized using chemical peptide synthesis methods or it can be synthesized by expression of a nucleic acid construct containing linked sequences encoding the antigenic and heat shock protein binding domains. One suitable technique utilizes initial separate PCR 20 amplification reactions to produce separate DNA segments encoding the two domains, each with a linker segment attached to one end, followed by fusion of the two amplified products in a further PCR step. This technique is referred to as linker tailing. Suitable restriction sites may also be engineered into regions of interest, after which restriction digestion and ligation is used to produce the desired hybrid antigen-encoding sequence. 25 [0069] As noted herein, the nucleic acid encoding a hybrid antigen of the invention is also suitable for therapeutic use by administration to the subject, where expression in vivo yields the hybrid antigen with the ability of inducing an immune response. Heat Shock Proteins [0070] The term "heat shock protein," as used herein, refers to any protein which 30 exhibits increased expression in a cell when the cell is subjected to a stress. In preferred 51 WO 2004/091493 PCT/US2004/010983 non-limiting embodiments, the heat shock protein is originally derived from a eukaryotic cell; in more preferred embodiments, the heat shock protein is originally derived from a mammalian cell. For example, but not by way of limitation, heat shock proteins which may be used according to the invention include BiP (also referred to as grp78), hsp70, hsc70, 5 gp96 (grp94), hsp60, hsp40, and hsp90, and members of the families thereof. Especially preferred heat shock proteins are BiP, gp96, and hsp70, as exemplified below. Most preferred is a member of the hsp70 family. Naturally occurring or recombinantly derived mutants of heat shock proteins may also be used according to the invention. For example, but not by way of limitation, the present invention provides for the use of heat shock 10 proteins mutated so as to facilitate their secretion from the cell (for example having mutation or deletion of an element which facilitates endoplasmic reticulum recapture, such as KDEL or its homologues; such mutants are described in PCT Application No. PCT/US96/13233 (WO 97/06685), which is incorporated herein by reference). [0071] For embodiments of the invention wherein heat shock protein and hybrid antigen 15 are directly administered to the subject in the form of a protein/peptide complex, the heat shock protein may be prepared, using standard techniques, from natural sources, for example as described in Flynn et al., Science 245:385-390 (1989), or using recombinant techniques such as expression of a heat shock encoding vector in a suitable host cell such as a bacterial, yeast or mammalian cell. If pre-loading of the heat shock protein with peptides 20 from the host organism is a concern, the heat shock protein can be incubated with ATP and then repurified. Non-limiting examples of methods for preparing recombinant heat shock proteins are set forth below. [0072] A nucleic acid encoding a heat shock protein may be operatively linked to elements necessary or desirable for expression and then used to express the desired heat 25 shock protein as either a means to produce heat shock protein for use in a protein vaccine or, alternatively, in a nucleic acid vaccine. Elements necessary or desirable for expression include, but are not limited to, promoter/enhancer elements, transcriptional start and stop sequences, polyadenylation signals, translational start and stop sequences, ribosome binding sites, signal sequences and the like. For example, but not by way of limitation, genes for 30 various heat shock proteins have been cloned and sequenced, including, but not limited to, gp96 (human: Genebank Accession No. X1 5187; Maki et al., Proc. Natl. Acad. Sci. US.A. 52 WO 2004/091493 PCT/US2004/010983 87:5658-5562 (1990); mouse: Genebank Accession No. M16370; Srivastava et al., Proc. Natl. Acad. Sci. U.S.A. 84:3807-3811 (1987)), BiP (mouse: Genebank Accession No. U16277; Haas et al., Proc. Natl. Acad. Sci. U.S.A. 85:2250-2254 (1988); human: Genebank Accession No. M19645; Ting et al., DNA 7:275-286 (1988)), hsp70 (mouse: Genebank 5 Accession No. M35021; Hunt et al., Gene 87:199-204 (1990); human: Genebank Accession No. M24743; Hunt et al, Proc. Natl. Acad. Sci. U.S.A. 82:6455-6489 (1995)), and hsp40 (human: Genebank Accession No. D49547; Ohtsuka K., Biochemn. Biophys. Res. Comnmun. 197:235-240 (1993)). METHODS OF ADMINISTRATION 10 [0073] The hybrid antigens of the invention or complexes of hybrid antigens and heat shock proteins may be administered to a subject using either a peptide-based, protein-based or nucleic acid vaccine, so as to produce, in the subject, an amount of complex which is effective in inducing a therapeutic immune response in the subject. [0074] The subject may bea human or nonhuman subject. 15 [0075] The term "therapeutic immune response," as used herein, refers to an increase in humoral and/or cellular immunity, as measured by standard techniques, which is directed toward the hybrid antigen. Preferably, but not by way of limitation, the induced level of humoral immunity directed toward hybrid antigen is at least four-fold, and preferably at least 16-fold greater than the levels of the humoral immunity directed toward the antigen 20 prior to the administration of the compositions of this invention to the subject. The immune response may also be measured qualitatively, by means of a suitable in vitro or in vivo assay, wherein an arrest in progression or a remission of neoplastic or infectious disease in the subject is considered to indicate the induction of a therapeutic immune response. [0076] Specific amounts of heat shock protein/hybrid antigen administered may depend 25 on numerous factors including the immunogenicity of the particular vaccine composition, the immunocompetence of the subject, the size of the subject and the route of administration. Determining a suitable amount of any given composition for administration is a matter of routine screening. 53 WO 2004/091493 PCT/US2004/010983 [0077] Furthermore, significant immunological efficacy was identified in studies in which the hybrid antigen was administered alone, i.e., without heat shock protein. While Applicants have no duty to disclose the theory by which the invention operates, and are not bound thereto, the results of these studies suggest that the hybrid antigens, upon injection 5 into the subject, bind to endogenous heat shock proteins, and thus do not require the concomitant administration of heat shock protein for effectiveness. The present invention extends to such utilities of the hybrid antigens of the invention, and moreover, to concomitant therapies or treatments that increase endogenous heat shock protein levels systemically or at the intended site of administration of the hybrid antigens of the invention. 10 Such concomitant therapies or treatments include but are not limited to local application of heat or local or systemic pharmaceutical agents that increase the expression of heat shock protein in the local tissue. Such agents and methods are known in the art. [0078] Hybrid antigens that are administered in the absence of co-administration of a heat shock protein (i.e., administered not in a complex with a heat shock protein) that 15 comprise at least one antigenic domain and at least one heat shock protein binding domain comprise one of the peptide linkers mentioned hereinabove. [0079] In specific non-limiting embodiments of the invention, it may be desirable to include more than one species of heat shock protein, and/or more than one hybrid antigen, 20 in order to optimize the immune response. Such an approach may be particularly advantageous in the treatment of cancer or in the treatment of infections characterized by the rapid development of mutations that result in evasion of the immune response. Moreover, a hybrid antigen of the invention may include more than one immunogenic domain or more than one epitope. 25 [0080] Compositions comprising hybrid antigen/heat shock protein or hybrid antigen alone as set forth above are referred to herein as "vaccines." The term vaccine is used to indicate that the compositions of the invention may be used to induce a prophylactic or therapeutic immune response. A vaccine of the invention may comprise a hybrid antigen with a single antigenic domain or epitope, or a hybrid antigen with a plurality of antigenic 30 domains or epitopes. Further, a vaccine may comprise an admixture of hybrid antigens with single or pluralities of antigenic domains or epitopes, or any combination of the foregoing. 54 WO 2004/091493 PCT/US2004/010983 As noted above, the hybrid antigens or admixtures thereof may be complexed with one or more heat shock proteins before administration, or may be administered without heat shock protein. [0081] A vaccine composition comprising one or more hybrid antigens optionally 5 complexed to one or more heat shock proteins in accordance with the invention may be administered cutaneously, subcutaneously, intradermally, intravenously, intramuscularly, parenterally, intrapulmonarily, intravaginally, intrarectally, nasally or topically. The vaccine composition may be delivered by injection, particle bombardment, orally or by aerosol. [0082] Incubation of heat shock proteins in solution with the hybrid antigen is sufficient 10 to achieve loading of the antigen onto the heat shock protein in most cases. It may be desirable in some cases, however, to add agents which can assist in the loading of the antigen. [0083] Incubation with heating of the heat shock protein with the hybrid antigen will in general lead to loading of the antigen onto the heat shock protein. In some cases, however, it 15 may be desirable to add additional agents to assist in the loading. For example, hsp40 can facilitate loading of peptides onto hsp70. Minami et al., J. Biol. Chemn. 271:19617-19624 (1996). Denaturants such as guanidinium HCl or urea can be employed to partially and reversibly destabilize the heat shock protein to make the peptide binding pocket more accessible to the antigen. 20 [0084] In particular, a vaccine of the invention comprising a heat shock protein preferably also includes adenosine diphosphate (ADP), to promote the association between the heat shock protein and the heat shock protein binding domain prior to the complex reaching its destination. Other compounds with similar capabilities may used, alone or in combination with ADP. 25 [0085] Vaccine compositions in accordance with the invention may further include various additional materials, such as a pharmaceutically acceptable carrier. Suitable carriers include any of the standard pharmaceutically accepted carriers, such as phosphate buffered saline solution, water, emulsions such as an oil/water emulsion or a triglyceride emulsion, various types of wetting agents, tablets, coated tablets and capsules. An example of an 30 acceptable triglyceride emulsion useful in intravenous and intraperitoneal administration of 55 WO 2004/091493 PCT/US2004/010983 the compounds is the triglyceride emulsion commercially known as Intralipid®. Typically such carriers contain excipients such as starch, milk, sugar, certain types of clay, gelatin, stearic acid, talc, vegetable fats or oils, gums, glycols, or other known excipients. Such carriers may also include flavor and color additives or other ingredients. 5 [0086] The vaccine composition of the invention may also include suitable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such compositions may be in the form of liquid or lyophilized or otherwise dried formulations and may include diluents of various buffer content (e.g., Tris-HC1, acetate, phosphate), pH and ionic strength, additives such as albumin or gelatin to prevent absorption to surfaces, detergents (e.g., 10 Tween 20, Tween 80, Pluronic F68, bile acid salts), solubilizing agents (e.g. glycerol, polyethylene glycerol), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite), preservatives (e.g., Thimerosal, benzyl alcohol, parabens), bulking substances or tonicity modifiers (e.g., lactose, mannitol), covalent attachment of polymers such as polyethylene glycol to the protein, complexing with metal ions, or incorporation of the material into or 15 onto particulate preparations of polymeric compounds such as polylactic acid, polyglycolic acid, hydrogels, etc. or onto liposomes, microemulsions, micelles, unilamellar or multilamellar vesicles, erythrocyte ghosts, or spheroplasts. Such compositions will influence the physical state, solubility, stability, rate of in vivo release, and rate of in vivo clearance. The choice of compositions will depend on the physical and chemical properties 20 of the vaccine. For example, a product derived from a membrane-bound form of a protein may require a formulation containing detergent. Controlled or sustained release compositions include formulation in lipophilic depots (e.g. fatty acids, waxes, oils). Also comprehended by the invention are particulate compositions coated with polymers (e.g. poloxamers or poloxamines) and coupled to antibodies directed against tissue-specific 25 receptors, ligands or antigens or coupled to ligands of tissue-specific receptors. Other embodiments of the compositions of the invention incorporate particulate forms protective coatings, protease inhibitors or permeation enhancers for various routes of administration, including intramuscular, parenteral, pulmonary, nasal and oral. [0087] As an alternative to direct administration of the hybrid antigen optionally 30 complexed with heat shock protein, one or more polynucleotide constructs may be administered which encode the hybrid antigen, optionally with heat shock protein, in 56 WO 2004/091493 PCT/US2004/010983 expressible form. The expressible polynucleotide constructs are introduced into cells in the subject using ex vivo or in vivo methods. Suitable methods include injection directly into tissue and tumors, transfecting using liposomes (Fraley et al., Nature 370:111-117 (1980)), receptor-mediated endocytosis (Zatloukal et al., Ann. NYAcad. Sci. 660:136-153 (1992)), 5 particle bombardment-mediated gene transfer (Eisenbraun et al., DNA & Cell BioL 12:792 797 (1993)) and transfection using peptide presenting bacteriophage (Barry et al, Nature Medicine 2:299-305 (1996). The polynucleotide vaccine may also be introduced into suitable cells in vitro which are then introduced into the subject. [0088] To construct an expressible polynucleotide, a region encoding the heat shock 10 protein and/or hybrid antigen is prepared as discussed above and inserted into a mammalian expression vector operatively linked to a suitable promoter such as the SV40 promoter, the cytomegalovirus (CMV) promoter or the Rous sarcoma virus (RSV) promoter. The resulting construct may then be used as a vaccine for genetic immunization. The nucleic acid polymer(s) could also be cloned into a viral vector. Suitable vectors include but are not 15 limited to retroviral vectors, adenovinis vectors, vaccinia virus vectors, pox virus vectors and adenovirus-associated vectors. Specific vectors which are suitable for use in the present invention are pCDNA3 (InVitrogen), plasmid AH5 (which contains the SV40 origin and the adenovirus major late promoter), pRC/CMV (InVitrogen), pCMU II (Paabo et al., EMBO J. 5:1921-1927 (1986)), pZip-Neo SV (Cepko et al., Cell 37:1053-1062 (1984)) and pSRa 20 (DNAX, Palo Alto, CA). [0089] Various methods for preparation of heat shock proteins and hybrid antigens are disclosed in WO9706821 and WO9922761, which are incorporated herein by reference in their entireties. [0090] In the following examples, and throughout the application amino acids may be 25 represented using their single-letter codes, as follows: [0091] A alanine [0092] C cysteine [0093] D aspartic acid [0094] E glutamic acid 30 [0095] F phenylalanine 57 WO 2004/091493 PCT/US2004/010983 [0096] G glycine [0097] H histidine [0098] I isoleucine [0099] K lysine 5 [00100] L leucine [00101] M methionine [00102] N asparagine [00103] P proline [00104] Q glutamine 10 [00105] R arginine [00106] S serine [00107] T threonine [00108] V valine [00109] W tryptophan 15 [00110] Y tyrosine [00111] The present invention may be better understood by reference to the following non-limiting Examples, which are provided as exemplary of the invention. The following examples are presented in order to more fully illustrate the preferred embodiments of the invention. They should in no way be construed, however, as limiting the broad scope of the 20 invention. EXAMPLE 1 [0100] A variety of hybrid antigens were prepared, each comprising a heat shock protein binding domain and a cancer antigen epitope or the model Class I H2-Kb epitope from ovalbumin, SIINFEKL. A peptide linker was included between the two domains. The heat 25 shock protein binding domains used in these experiments were among the following: HWDFAWPW, NLLRLTGW, FYQLALTW and RKLFFNLRW. Linkers were among those desceibed hereinabove. [0101] The cancer and model epitopes were among the following: 58 WO 2004/091493 PCT/US2004/010983 Trivial Name Source Protein Source Tumor Amino Acids (Amino acid sequence) Prostate Specific Membrane PSMA P2 Antigen Prostate cancer 771-779 (ALFDIESKY) Antigen (ALFDIESKV) Gp 100 Melanoma 209-217 DID (21 OM) (IMDQVPFSV) Tyrosinase Melanoma 368-376 YD (370D) (YMDGTMSQV) Human Papillomavirus Cervical cancer 8693 HPVI6 E7 86-93 (HPV) Strain 16 E7 rvicaancer (TLGIVCPI) HPV16 E71 1-20 HPV Strain 16 E7 Cervical cancer 11-20 HPV16 E711-20 (YMLDLQPETT) Ovalbumin Model Tumor 257-264 Ova Antigen (SINFEKL) Using standard solid phase peptide synthesis using F-moc chemistry, hybrid antigens comprising a heat shock protein binding domain, a cancer epitope, and a linker there between, were synthesized, in various orientations. 5 EXAMPLE 2 [0102] Binding affinities between recombinant human or murine heat shock protein 70 (hsp70) and the various heat shock protein binding domains and antigenic peptides mentioned above, as well as between the hybrid antigens comprising an antigenic peptide 10 and a heat shock protein binding domain described above, were determined by a binding inhibition assays (Hill plots) relative to the binding affinity of a reference, labeled hybrid antigen (tritiated or fluoresceinated ALFDIESKVGSGHWDFAWPW) to hsp70 as determined by Scatchard analysis (Kds of 22.64 gM and 10.75 JIM, respectively). Binding studies were performed in 39% PBS; 20 mM THAM, pH 8; 37 mM NaCI, 5 mM MgC1 2 ; 15 and 1 mM ADP. EXAMPLE 3 For immunological studies in mice, a murine MHC H2-K(b) epitope from ovalbumin, SIINFEKL (amino acids 257-264), and a H2-K(b) peptide from the nucleoprotein of vesicular stomatitis virus (VSV), RGYVYQGL (amino acids 52-59) were used for the 59 WO 2004/091493 PCT/US2004/010983 preparation of hybrid antigens. The following table sets forth the sequences and the affinities for hsp70 of the epitopes alone and in hybrid antigens. Epitope alone Hybrid antigen comprising epitope Mouse Affinity Affinity Epitope Epitope for hsp70 forHybrid antigen equene hp7 sequence Hybrid antigen sequence hp70 (~Im) (I) NLLRLTGWGSGSIINFEKL 1.6 Ovalbumin: amino acids SIINFEKL 235 NLLRLTGWFFRKSIINFEKL 2.2 257-264 NLLRLTGWRKSIINFEKL 0.8 VSV nucleo- NLLRLTGWGSGRGYVYQGL 1.4 protein: RGYVYQGL 82 NLLRLTGWFFRKRGYVYQGL 1.0 amino acids 52-59 NLLRLTGWRKRGYVYQGL 0.6 EXAMPLE 4 5 [0103] Mice were immunized s.c. at the base of the tail with hsp70 alone, hsp70 complexed with SIINFEKL, and hybrid SIINFEKL peptide with or without HSP70. The doses were adjusted such that each immunization contained the same amount of SIINFEKL, except for hsp70 alone. Seven days later, spleens were harvested and enriched for CD8+ T cells, which were put into an ex vivo IFN-y ELISPOT assay. Responses after pulsing with 10 SIINFEKL ("SIINFEKL") were recorded in the following table, which includes the doses, and the number of spots (mean d- standard error) per 4 x 105 CD8 T cells, of> four experiments with at least three mice per group. Controls included medium alone ("medium control"), unpulsed T cells ("unpulsed control"), T cells pulsed with a non-immunized peptide derived from VSV, RGYVYQGL ("VSV control"), and exposure to concanavalin A 15 as a positive control ("Con A positive control"). 60 WO 2004/091493 PCT/US2004/010983 In the same experiment, a slCr-release assay as described above was done using SIINFEKL pulsed target cells. At an effector to target cell ratio of 200:1, the percent killing results obtained are shown in the far right column of the following table. (200-10) Number of Spots per 400,000 cells CTL assay: lmmunogen Con A SIINFEKL Medium Unpulsed VSV % killing at SIINFEL control control control poStl 200:1 E/T control 200:1 E/T 4.4 pgHsp70 0.00d0.00 1.50±2.12 0.6710.58 0.33±0.58 834±28.3 0% 4.4 pg Hsp70 + 0.9 gg 33.7 ± 7.09 0.00±0.00 0.33±0.58 0.00±0.00 1000±33.7 19% SIINFEKL 4.4 jg Hsp70 + 2.0 gg 80.0 ±17.0 0.0010.00 1.50±0.71 1.50±0.71 1170±56.5 38% NLLRLTGWGSGSIINFEKL 4.4 pg Hsp70 +2.4 gg 222 ± 17.7 0.00±0.00 0.67±0.58 1.33±1.53 1010156.5 52% NLLRLTGWFFRKSIINFEKL 5 EXAMPLE 5 [0104] An experiment similar to that described above was carried out, which also included hybrid antigen without hsp70. 61 WO 2004/091493 PCT/US2004/010983 (200-11) Number of Spots per 4 x 105 CD8 T cells Immunogen Con A SIINFEKL Medium Unpulsed VSV control Poitive control control VSV control Positive control 4.4 jig Hsp70 0.331 0.58 1.00 1.73 1.67 1 1.15 4.00 -1.00 965±62.6 4.4 tg Hsp70 + 0.9 ag 1.67 ± 0.58 1.00 + 1.00 2.00 + 0.00 2.67 + 2.08 591 ±48.1 SIINFEKL 4.4 gg Hsp70 + 2.0 jtg 12.0±5.2 2.67 : 0.58 1.67 ± 1.15 2.00± 2.65 748 58.6 NLLRLTGWGSGSIINFEKL 4.4 pg Hsp70 + 2.4 jig 770 ± 80.6 3.33 1 1.53 3.67 1 1.53 4.33 ± 1.53 742 - 72.6 NLLRLTGWFFRKSIINFEKL 2.4 gg 151 ± 20.7 1.00 1.00 1.67 ± 0.58 0.00 ± 0.00 459 E 149 NLLRLTGWFFRKSIINFEKL (no hsp70) EXAMPLE 6 A further experiment was carried out similar to that described above. 62 WO 2004/091493 PCT/US2004/010983 (200-12) Number of spots per 300,000 CD8 T cells CTL assay: Immunogen CnA %kliga SIINFEKL Medium Unpulsed VSV Con 200A % killing at control control control positive 200:1 ET control 4.4 ig Hsp70 0.67 -0.50 0.00 0.00 0.50 ± 0.71 1.00± 1.41 552 ± 24.0 0.45 41.3 4.4 gg Hsp70 + 0.9 3.33 + 2.52 0.00 + 0.00 0.33 0.58 0.33 : 0.58 450 ± 69.0 43.0 ± 21.2 jtg SIINFEKL 4.4 gg Hsp70 +2.00 134- -4.16 1.33 1.53 0.67± 1.15 1.00± 1.00 865+93.0 31.9±5.41 jig
NLLRLTGWGSG
SIINFEKL 4.4 gig Hsp70 + 2.4 680 + 23.0 0.00 ± 0.00 0.00 + 0.00 1.67 ± 0.58 801 J= 56.6 84.6 - 1.70 jIg
NLLRLTGWFFRK
SIINFEKL 2.4 gg 211 L 17.0 0.00 0.00 0.50±0.71 1.00 0.00 688 + 41.7 9.91- 5.57
NLLRLTGWFFRK
SIINFEKL EXAMPLE 7 [0105] As in the prior in vivo experiments, B6 mice were immunized s.c. to evaluate 5 complexes of hsp70 with hybrid antigens made using other short peptide linkers, including (using one-letter amino-acid codes) FFRK, RK, AKVL, QLK and FR, and at different doses. An ex vivo IFN-y ELISPOT assay was performed as described above. The results including the control values are as follows. 63 WO 2004/091493 PCT/US20041010983 (200- 13) Number of Spots per 300,000 cells I "RgnS[IIIFEKSL Medium Unpuled ~ VS control control control 4.4 pg Hsp70 + 2.4pg 114: 21 1.0kL 1.2 1.0 ± 0 0.67 =O .41 NLLRLTGWFFRKSIINFEKL 4.4pg Hsp7 + 2.4pg 70 ±8.5 1.3k 1.1 0.67 JO .82 2.7 =1 1. 1 NLLRLTGWRKSIINFEKL 0.9 tg Hsp70 +0.48gig 98:1:0.41 .67:60.82 1.3 ± 1. 1 4.3:12.3 NLLRLTGWFFRKSIINFEKL 0.9 p.g Hsp70 +0.48gg 29:b2.2 0 40 1:10 0 Ar0 NLLRLTGWRKSIINFEKL 2.4jtg 11 ±1.8 0.67:L0.82 D=E0 0.67 =O .82 NLLRLTGWFFRKSIINFEKL 200-21 Number of Spots per 400,000 cells Imniunogen SINEL Medium Unpulsed VSV control SIINEKL control control 4.4 jig Hsp7O + 2.4jig 124 1 8.8 0.33 ± 0.41 0.67 ± 0. 82 2.67 ± 2.68 NLLRLTGWFFRKSIINFEKL 4.4 jIg Hsp7 + 2.4jig 95± 12 1.33 4-0.82 LO0± 1.2 0.67+±0.41 NLLRLTGWAKVLSIINFEKL 64 WO 2004/091493 PCT/US2004/010983 200-23 Number of Spots per 400,000 cells Immunogen SHFNFEI L Medium Unpulsed VSV control control control 4.4 pgg Hsp70 + 2.4 gg 318+17 0.67 0.51 0.67 ± 0.58 0.67 ± 0.58 NLLRLTGWFFRKSTINFEKL 4.4 gig Hsp70 + 2.4 gg 174 ± 18 0.0 & 0.0 0.0 : 0.0 3.7 & 2.5 NLLRLTGWQLKSIINFEKL 4.4 pg Hsp70 + 2.4 Rg 53 ± 2.9 0.0 & 0.0 0.67 ± 0.58 1.0 1.0 NLLRLTGWFRSIINFEKL 2.4 pg 31 4 5.7 1.0 =- 1.7 0.0 = 0.0 0.67 + 0.58 NLLRLTGWFRSIINFEKL EXAMPLE 8 Similar in vivo studies in B6 mice as those described above were performed using 5 formulations without added hsp70. The results are as follows. 65 WO 2004/091493 PCT/US2004/010983 (200-17) Number of Spots per 400,000 cells Immunogen Con A SIIN\FEKL control control VSV control positive control 10 pg SIINFEKL 2.33 + 0.41 0,33 : 0.41 1.33 : 0.82 1.7 + 0.41 928 72 0.5 pg 22+ 7.2 1.33 ± 0.41 1.67+ 1.1 1.0± 0.71 906: 17 NLLRLTGWFFRKSIINFEKL 2.5 pg 28 2.7 1.0 1.7 0.33 0.41 2.0 -- 1.2 930 ± 23 NLLRLTGWFFRKSIINFEKL 25 pg 46 1 4.3 2.01+0.41 1.33 1.1 3.0 ± 0.71 1007 17 NLLRLTGWFFRKSIINFEKL EXAMPLE 9 Similar in vivo studies in B6 mice as those described above were performed using 5 formulations with or without hsp70. In addition, one study was carried out in which hybrid antigen was co-administered with free heat shock protein-binding domain peptide (NLLRLTGW). The results are as follows. 66 WO 2004/091493 PCT/US2004/010983 (VSV-72-02) Number of spots per 400,000 cells CTL Immunogen % SIINFEKL Medium Unpulsed VSV Con A positive killing control control control control at 200:1 E/T 4p.g Hsp70 + 2.0tg 48+11 0.0± 1.0 0.0± 1.0 4.0 + 2.0 588+ 151 32% NLLRLTGWFFRKSIINFEKL 2.0ltg 24+1 1.0 ± 1.0 1.0 ± 1.0 5.0 + 3.0 842 + 73 24% NLLRLTGWRKSIINFEKL 2.0 jig 2.0- 1.0 1.0 ± 1.0 0.0± 1.0 1.04 1.0 422: 54 18% NLLRLTGWFFRKSIINFEKL + 50-fold excess NLLRLTGW SIINFEKL 1.0. 4 1.0 0.0 +40.0 0.0-+ 1.0 1.0+- 1.0 478 4 67 6% EXAMPLE 10 The VSV epitope used as a control in many of the foregoing experiments, RGYVYQGL, 5 was used as the epitope in preparing further hybrid antigens of the invention, and evaluated for induction of an immune response in similar experiments as described above. 67 WO 2004/091493 PCT/US2004/010983 (VSV-72-02) Number of Spots per 400,000 cells Immunogen VSV Medium Unpulsed SllNfFEK L control control (RGYVYQGL) 4gg HSP plus 2pg 48= 11 1.0± 1.0 0.0 1 1.0 4.0 12.0 NLLRLTGWFFRKSIINFEKL 4lig HSP plus 2pg 1.0 11.0 1.0 + 1.0 4.0 ± 2.0 20 - 1.0 NLLRLTGWFFRKRGYVYQGL 4Rg HSP plus 61.tg 6.0 - 3.0 2.0 ± 2.0 12 ± 3.0 104 -± 13 NLLRLTGWFFRKRGYVYQGL EXAMPLE 11 In order to evaluate the efficacy of the aforementioned hybrid antigens and complexes with 5 hsp70 on the treatment of disease, a model was utilized in which 20,000 E7 tumor cells modified to express ovalbumin (designated E.G7) were subcutaneously implanted in B6 mice. Ten mice were used per treatment group. This model is described, for example, in Moroi et al., 2000, Proc. Nat. Acad. Sci. USA 97:3485-3490. The results in number of mice with tumors over time are shown in Figure 1. After 31 days, none of 10 mice immunized 10 with hsp70:NLLRLTGWFFRKSIINFEKL developed tumors, nor did mice immunized with SIINFEKL emulsified in Titermax adjuvant. Three of 10 mice vaccinated with NLLRLTGWFFRKSIINFEKL alone (no hsp70) had tumors. Five of 10 mice vaccinated with hsp70: SIINFEKL had tumors, and 9 of 10 mice immunized with Titermax and buffer alone had tumors. 15 EXAMPLE 12 The in-vitro antigen presentation assay described above was utilized further in order to evaluate the formulations of the invention. To demonstrate the requirement of the hybrid antigens of the invention for hsp70, whether supplied in the formulation or endogenously 68 WO 2004/091493 PCT/US2004/010983 available, for entry of the hybrid antigen and more specifically its antigen into the antigen presentation pathway, the assay was performed with the following formulations, with the results indicated. (200-MF-41) Formulation Pg/ml IL-2 produced by B3Z cells 0.5 ng SIINFEKL 2690 : 369 5 ng NLLRLTGWFFRKSIINFEKL 46 : 11 5 ng NLLRLTGWFFRKSIINFEKL 3920 ± 344 plus 1.4 ug hsp70 1.4 ug Hsp70 0.0 = 0.0 5 EXAMPLE 13 [0106] The HHD II mouse model bearing a human HLA-A2 complex described by Firat et al., 1999, "H-2 class I knockout, HLA-A2.1 -transgenic mice: a versatile animal model for preclinical evaluation of antitumor immunotherapeutic strategies," Eur Jlmmunol. 29:3112 10 21, was used in the following experiments to evaluate human HLA-A2 epitopes in hybrid antigens of the invention. The "IMD" peptide epitope IMDQVPFSV from the human melanoma antigen gp 100 was evaluated in a hybrid antigen of the invention at low and high dose in the HHD II model. Similar methods to those described above were used for the ELISPOT assay, with test peptides being the IMD peptide and, as a control, a peptide from 15 the melanoma antigen tyrosinase, YMDGTMSQV ("YMD"). The results are shown on the following table. 69 WO 2004/091493 PCT/US2004/010983 (HHD It 200-72-02) Number of Spots per 400,000 cells Immunogen IMID Medium control Unpulaed control YMVID control 4pg hsp70 and 5pg 139+11 0.67 0.58 1.0 ± 1.0 3.7 0.58 NLLRLTGWFFRKIMDQVPFSV 4gg hsp70 and 10pg 217 3.2 0.67k 0.58 4.0±6.0 2.7± 1.5 NLLRLTGWFFRKIMDQVPFSV 2pg 27h 5.1 0.0± 0.0 0.0 ± 0.0 2.0±2.0 NLLRLTGWFFRKIMDQVPFSV A similar experiment in HHD II mice carried out using YMD as the epitope in the hybrid antigen, in a complex with hsp70, as follows. 5 (200-72-01) Number of Spots per 400,000 cells Immunogen YMD Medium control Unpulsed control IMD control 4pg hsp70 and 5gg 33 ± 7.8 1.0 ± 0.0 1.0 1 0.0 1.0 ± 1.4 NLLRLTGWFFRKYMDGTMSQV 4pg hsp70 and 10gg 323144 0.0 0.0 1.510.71 1.5 0.71 NLLRLTGWFFRKYMDGTMSQV 70 WO 2004/091493 PCT/US2004/010983 EXAMPLE 14 An epitope from Sendai virus (SdV), FAPGNYPAL, was evaluated in hybrid antigens of the invention in B6 mice, similar to the above. The results are as follows. (200-18) Number of Spots per 400,000 cells Immunogen Medium SIINFEKL SdV control control 4tg hsp70 and 2.g 1.3± 1.2 1.0± 1.0 197± 27 NLLRLTGWFFRKSIINFEKL 2ptg 0.33±0.58 0.0± 0.0 87±20 NLLRLTGWFFRKRGYVYQGL 4pg hsp70 and 2!g 38 ± 17 0.33 ± 1.0 ± 1.0 NLLRLTGWFFRKFAPGNYPAL 0.58 13pg hsp70 and 7tg 169 : 32 4.3 - 1.5 7.0 ± 3.5 NLLRLTGWFFRKFAPGNYPAL 5 EXAMPLE 15 In-vivo experiments on co-administration of two hybrid antigens of the invention with hsp70 to B6 mice was performed. Hybrid antigens containing SIINFEKL from ovalbumin and RGYVYQGL from VSV were admixed and immunized with hsp70. The results are as 10 follow. 71 WO 2004/091493 PCT/US2004/010983 (OVA-VSV-72-01) Number of Spots per 400,000 cells Immunogen VSV Medium Unpulsed OVA control control 24g hsp70 77 19 2.0± 1.0 2.0k 1.0 366 - 19 2ptg NLLRLTGWFFRKSIINFEKL 2gg NLLRLTGWFFRKRGYVYQGL 2gg hsp70 185 ± 9 1.0- 1.0 4.0 ± 2.0 349 1 10 6pg NLLRLTGWFFRKSIINFEKL 64g NLLRLTGWFFRKRGYVYQGL EXAMPLE 16 As noted above, in one aspect of the invention, formulations containing a plurality of hybrid 5 antigens comprising different antigenic epitopes may be formulated with one or more heat shock proteins for immunization in humans in order to elicit an effective immune response to treat or prevent a disease. For example, for treating human melanoma, a formulation comprising 8 different melanoma epitopes may be prepared as hybrid antigens, and formulated, for example, with hsp70. In this particular formulation, the heat shock protein 10 binding domain NLLRLTGW at the N-terminus is used for all epitopes, linked to the epitope at the C-terminus using the peptide linker FFRK. Other binding domains and linkers are embraced herein. This particular formulation is useful for treating patients with the HLA-A2 haplotype. A formulation comprises the following hybrid antigens with hsp70: 72 WO 2004/091493 PCT/US2004/010983 Source and amino acid sequence of Hybrid antigen sequence antigen gp 100: amino acids 209-217 (modified NLLRLTGWFFRKIMDQVPFSV 210M) tyrosinase: amino acids 368-376 (modified NLLRLTGWFFRKYMDGTMSQV 370 D) Melan-A: amino acids 26-35 (modified 27L) NLLRLTGWFFRKELAGIGILTV NY-ESO-1: amino acids 157-165 (modified NLLRLTGWFFRKSLLMWITQV 165V) TRP-2: amino acids 180-188 NLLRLTGrFFRKSVYDFFVWL MAGE- 10: amino acids 254-262 GLYDGMEHLGSGNLLRLTGW gpl00: amino acids 280-288 (288V) YLEPGPVTVGSGNLLRLTGW SSX-2: amino acids 41-49 KASEKIFYVGSGNLLRLTGW In one embodiment, approximately equal amounts of the foregoing 8 hybrid antigens may be complexed with hsp70, and administered in saline. In another embodiment, a 5 formulation comprises the first five hybrid antigens listed. The aforementioned formulations containing heat shock protein in saline optionally may contain ADP to stabilize the complexes, as well as other components, such as excipients, diluents and carriers, as mentioned above. In another embodiment, an admixture of the foregoing 8 hybrid antigens, or the first 5 listed, is formulated in saline for administration without a heat 10 shock protein. 73 WO 2004/091493 PCT/US2004/010983 EXAMPLE 17 Prime-boost protocols were valuated in this experiment. Using the NLLRLTGWFFRKSIINFEKL hybrid antigen, or without co-administered hsp70, the following 5 protocols were followed: 1) administer at day 0, analyze at day 7; 2) 5 administer at days 0 and 7, analyze at day 21; 3) administer at day 0, analyze at day 21; 4) administer at days 0 and 14, analyze at day 28; and 5) administer at day 0 and analyze at day 28. The results in number of spots per 400,000 cells, were as follows. (200-28-72-0la, -01b, -1Oe) Protocol day(s) immunized 0 0, 7 0 0, 14 0 Protocol day analyzed 7 21 21 28 28 SIINFEKL 3.0± 2.0 2.0± 1.0 0.0±+ 1.0 1.0± 1.0 .0 ± 1.0 2gg hsp70, 41g SIINFEKL 3.0 ± 1.0 6.0 ± 1.0 22 ± 8.0 3.0 ± 2.0 20 ± 5.0 2pg 72 ± 5.0 24 ± 6.0 42+7.0 25 9.0 82± 11 NLLRLTGWFFRKSIINFEKL 2gg hsp70 and 4jg 99±12 98±11 141±14 398 18 27± 2.0 NLLRLTGWFFRKSIINFEKL 2pg hsp70 5.0 4 6.0 3.0 : 2.0 3.0 ± 0.0 1.0 = 1.0 4.0 3.0 10 EXAMPLE 18 Further experiments were performed with mixtures of hybrid antigens to demonstrate eliciting of an immune response to the component antigens, as above. In this experiment, hybrid antigens containing SIINFEKL and the VSV peptide RGYVYQGL were used. 74 WO 2004/091493 PCT/US2004/010983 (VSV/OVA-72-02) Number of Spots per 300,000 cells Immunogen SIIFEKL Medium Unpulsed VSV control control (RGYVYQGL) 3.7 jig hsp70 238 27 0.0 ± 0.0 1.0 1.0 5.0 4 2.0 2pg NLLRLTGWFFRKSIINFEKL 11.2 gg hsp70 330 45 1.0 : 1.0 0.0 A 0.0 4.0 ± 1.0 6pg NLLRLTGWFFRKSIINFEKL 3.7 pg hsp70 1.0 ±1.0 1.0 ± 1.0 0.0 ± 0.0 61 11 2pg NLLRLTGWFFRKRGYVYQGL 11.2 pg hsp70 2.0 b 2.0 2.0 1.0 2.0 0.0 147 ± 20 6pg NLLRLTGWFFRKRGYVYQGL 3.7 pg hsp70 179 4.0 2.0 12.0 1.0 1.0 165± 11 2pg NLLRLTGWFFRKSIINFEKL 2tg NLLRLTGWFFRKRGYVYQGL 11.2 pig hsp70 310 -13 1.0 - 1.0 1.0 4 1.0 242 -52 6pg NLLRLTGWFFRKSIINFEKL 6gg NLLRLTGWFFRKRGYVYQGL EXAMPLE 19 The binding affinity for hybrid antigens comprising heat shock protein binding domain 5 NLLRLTGW, antigenic domain SIINFEKL (from ovalbumin) or RGYVYQGL (from VSV protein) and various linkers set forth in Example 32 were carried out as described in Example 17. The antigenic domains alone had a Kd for hsp70 binding of 235 pM and 82 pM, respectively. The results are shown below. 75 WO 2004/091493 PCT/US2004/010983 Hybrid antigen Kd for binding to HSP70 NLLRLTGWGSGSIINFEKL 1.6 1M NLLRLTGWFFRKSIINFEKL 2.2 pM NLLRLTGWRKSIINFEKL 0.8 piM NLLRLTGWAKVLSIITNFEKL 2.0 pM NLLRLTGWQLKSIlNFEKL 0.4 pIVIM NLLRLTGWFRSIINFEKL 1.5 pAM NLLRLTGWGSGRGYVYQGL 1.4 pM NLLRLTGWFFRKRGYVYQGL 1.0 piM NLLRLTGWRKRGYVYQGL 0.6 pIM EXAMPLE 20 5 Further studies were carried out to evaluate the immunogenicity of hybrid antigens when administered alone to B6 mice, without co-administration of hsp70. The methods for evaluation using IFN-y ELISPOT are as described above. 76 WO 2004/091493 PCT/US2004/010983 (Control 200-24 and 200-30) Number of Spots per 300,000 cells Immunogen SIINFEKL SWDFITV Medium Unpulsed Splenocytes 25 ig £1T 0 4-0 3.0 4-2.0 NLLRLTGWFFRKSIINFEKL 109 4 14 NT 0 0 3.02.0 24.9 gg NLLRLTGWFFRKSSWDFITV NT 26 5 0.67 ± 0.58 0.33 ± 0.58 2.1 pg NLLRLTGWFRSINFEKL 12 -2 NT 0.67 L 0.58 0.67 + 0.58 NT not tested EXAMPLE 21 5 Hybrid antigens were prepared comprising two antigens, separated by a linker as described above, such that the hybrid antigen has the following general structure: (Heat shock protein binding domain) - (linker) - (Antigen 1) - (linker) - (Antigen 2). While in this example the heat shock protein binding domain is at the N-terminal portion of the hybrid antigen, this is not necessarily the case and hybrid antigens with the heat shock 10 protein binding domain at the C-terminus, or in-between the two antigenic domains, is embraced by the present invention. Furthermore, although in the examples below the same linker peptide is used between the antigenic domains and between the antigenic domain proximal to the heat shock protein binding domain, this is not necessarily the case and different linker peptides may be used. Moreover, the presence of the linker in one or both 15 positions is optional. And furthermore, three or more antigenic peptides may be used. For simplicity, such hybrid antigens with two or more antigenic domains is termed a tandem hybrid antigen. Such tandem hybrid antigen compositions, complexes of one or more tandem hybrid antigens and a heat shock protein, and methods of eliciting an immune response or preventing or treating a disease by administering one or more tandem hybrid 20 antigens or complexes of at least one heat shock protein and at least one tandem hybrid antigen are fully embraced herein. 77 WO 2004/091493 PCT/US2004/010983 The following experiments compare the immunogenicity of the admixture of two hybrid antigens and a tandem hybrid antigen comprising the same antigens, and a dose response study. In one experiment, a peptide comprising two linkers and epitopes but no heat shock protein binding domain was included. 5 (Control-200-72-01) Number of Spots per 300,000 cells Immunogen SIINFEKL Medium Unpulsed RGYVYQGL control control 19.2 g 390+56 1.7± 1.1 3.0 41.9 146:613 NLLRLTGWFFRKSIINFEKLFFRKRGYVYGL 19.2 jig 180E11 1.3 +1.1 2.7 ± 1.1 321-5.8 NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL (S200-72-02) Number of Spots per 300,000 cells Immunogen SIINFEKL Medium control RGYVYQGL 7.3 pg 8.3 1.1 1.7 4- 0.4 31 ± 5.5 FFRKSIINFEKLFFRKRGYVYQGL 9.6 pg 713 ± 13 9.0 1.2 207 8.2 NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 9.6 pg 69-12 0.7-0.4 460114 NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL 78 WO 2004/091493 PCT/US2004/010983 (S200-72-12) Number of Spots per 300,000 cells Immunogen SIINFEKL Medium control RG'VYQGL 20 pg 410 49 0.3 "0.4 250± 11 NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 10 jgg 360 ± 13 0.3 .0.4 100 110 NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 5 lig 130 ±3.3 0. 0 35 4 6.6 NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 20 gg 150 . 6 0 0 380 12 NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL 10ig 30-3 00 83±5 NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL In this and other experiments, the epitope proximal to the heat shock protein binding domain exhibited the strongest immune response, and thus the positioning of the selected 5 epitopes selected for the vaccine formulations of the invention may be positioned to contribute maximally to the overall immunogenicity of the formulation, whether administered in the absence of heat shock proteins or administered as complexes with heat shock proteins. EXAMPLE 22 10 In the following experiments, admixtures of tandem hybrid antigens were evaluated for immunogenicity. In addition to the H2-Kb Class I peptides from ovalbumin (SIINFEKL) and from VSV (RGYVYQGL), the H2-Kb j3-casein peptide IAYFYPEL and the Sendai virus peptide FAPGNYPAL were also used. In another experiment, two tandem hybrid antigens with the same antigenic peptides in alternate configurations were admixed. Strong 15 immune responses to four epitopes were elicited. 79 WO 2004/091493 PCT/US2004/010983 All of the formulations herein included 1 mM ADP. In one experiment described below, ADP was omitted. (200-72-04) Number of Spots per 300,000 cells Immunogen SIINFEKL RGYVYQGL IAYFYPEL FAPGNYPAL 9.6 gg 537 16 150 10 4.7: 0.8 5.7 12.5 NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 9.7 Fg 1.7 ±1.1 1.7 0.8 128 4 9.2 136 ± 6.6 NLLRLTGWFFRKIAYFYPELFFRKFAPGNYPAL 9.6 gg NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL plus 363 ± 31 256 ± 5.3 127 4 7.9 155 ± 28 9.7 gg NLLRLTGWFFRKIAYFYPELFFRKFAPGNYPAL 5 S200-72-13 Number of Spots per 300,000 cells Immunogen SHNFEKL RGYVYQGL IAYFYPEL FAPGNYPAL 9.6 pg NLLRLTGWFFRKIAYFYPELFFRKFAPGNYPAL plus 388 ± 6.8 72 ± 5.0 402 ± 17 379 ± 30 9.6 pg NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 9.6 pg NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL Plus 76 ±1.9 159 ± 8.3 115 20 172 5.9 9.6 pg NLLRLTGWFFRKIAYFYPELFFRKFAPGNYPAL 80 WO 2004/091493 PCT/US20041010983 S200-72- 13 N umber of Spots per 300,000 cells Immunogen SUNFEKL RGYVYQGL IAYFYPEL Medium 9.6 jig4510 231 3014 0.3.4 NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 45±0 -73L2 .064 031.1 9.6 pg 82 ±4 445 ±30 1.3 40.41 0±0 NLLRLTGjWFFRKRGYVYQGLFFRKISINFEKL 9.6 pg NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL plus 202:6 7.6 188 1 24 1.0± 0.7 0.67 +0.41 9.6 pg NLLRLTGWFFRKRGYVYQGLFFRKJSIINFEYJL S200-72-13, no ADP Number of Spots per 300,000 cells Immunogen SIINFEKL RGYVYQGL IAYFYPEL Medium 9.6 gg 22812.5 12612.9 1.7 ±0.4 0±0 NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 9 .6 Vg 83 :L9 189119 13 415 0.33 ±0.41 NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL 9.6 p~g NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL plus 115±7.8 86:E11 0.33± 0.41 010 9.6 pg NLLRLTGWFFRKRGiYVYQGLFFRKSINFEKL 5 WO 2004/091493 PCT/US2004/010983 EXAMPLE 23 In the following experiment, up to five antigenic peptides are delivered and induce immunogenicity without co-administered HSP70, when administered as an admixture of two tandem hybrid antigens and a single hybrid antigen to B6 mice. The tandem hybrid 5 antigens included VSV and ovalbumin peptides in one, and P-casein and Sendai virus peptides in the other. The single hybrid antigen contained NS2-114 influenza peptide (RTFSFQLI). S200-72-15 Number of Spots per 300,000 cells Immunogen SIINFEKL RGYVYQGL IAYFYPEL FAPGNYPAL RTFSFQLI 9.6 gg NLLRLTGWFFRKRGYVYQGL FFRKSIINFEKL plus 67 L 6.1 205± 20 229 ± 28 266 -33 0 1 0 19 Ag NLLRLTGWFFRKIAYFYPELF FRKFAPGNYPAL 9,6 pg NLLRLTGWFFRKRGYVYQGL FFRKSIINFEKL plus 19 pg 156-3.3 299 ± 18 175 12 125 3.3 33 4.7 NLLRLTGWFFRKIAYFYPELF FRKFAPGNYPAL plus 12.2 4g NLLRLTGWFFRKRTFSFQLI 10 EXAMPLE 24 The immunogenicity of the foregoing single hybrid antigens administered without heat shock protein were evaluated in combination with helper T cell epitopes present in a hybrid antigen. In most experiments, a H2-Kb Class II epitope from ovalbumin, amino acids 323 339, TEWTSSNVMEERKIKV, was used (i.e., the hybrid antigen had a sequence of 82 WO 2004/091493 PCT/US2004/010983 NLLRLTGWFFRKTEWTSSNVMEERKIKV). Inclusion of the Class II peptide containing hybrid antigen increased the response to the Class I epitope on the average of about seven fold. (250-72-08) Number of Spots per 300,000 cells Response to Response to Class I Class I hybrid Class I epitope when peptide-containing epitope Class I and immunogen when Class I Class II Medium Splenocytes hybrid hybrid antigen antigen administered admixture is administered 24.2 ig 2 + 1.9 13 -3.9 0.7 : 0.4 0 0 NLLRLTGWFFRKDAPIYTNV 24.9 gg 18 ± 0.7 98 ± 5.8 0.7 + 0.8 0.7 0.4 NLLRLTGWFFRKSSWDFITV 25.4 gg 5.3 1.5 43 ± 7.6 0.3 ± 0.4 0 ± 0 NLLRLTGWFFRKRTFSFQLI 25.5 gg 1113 7319.8 0±0 0-0 NLLRLTGWFFRKIAYFYPEL 5 EXAMPLE 25 The effect on immunogenicity of hybrid antigens co-administered with various hybrid antigens containing H2-Kb Class II peptides, in the absence of heat shock protein, were evaluated. The Class I peptides were either SSWDFITV or DAPIYTNV; Class II peptides 10 included the ovalbumin peptide mentioned above, a Class II peptide from tetanus toxoid NNFTVSFWLRVPKVSASHL (i.e., the hybrid antigen has a sequence of NLLRLTGWFFRKNNFTVSFWLRVPKVSASHL), or a HBVc (amino acids 128-140) peptide, TPPAYRPPNAPIL. 83 WO 2004/091493 PCT/US2004/010983 250-72-13 Number of Spots per 300,000 cells Immunogen Medium SSWDFITV 24.9 pg 3.0 0.7 78 h 3.9 NLLRLTGWFFRKSSWDFITV 24.9 gg 8.0 -3.1 NLLRLTGWFFRKSSWDFITV plus 84 ± 7.1 27.4 gg NLLRLTGWFFRKTPPAYRPPNAPIL 24.9 gg NLLRLTGWFFRKSSWDFITV plus 3.7 1.1 315 j 15 33.6 gg NNFTVSFWLRVPKVSASHLGSGNLLRLTGW 24.9 pg NLLRLTGWFFRKSSWDFITV plus 2.7 1 2.0 135± 5.7 36.4 pg HWDFAWPWNGSGNNFIVSFWLRVPKVSASHL 24.9 tg NLLRLTGWFFRKSSWDFITV plus 1.7 ± 0.4 229 12 34.7 ag NLLRLTGWFFRKTEWTSSNVMEERKIKV Thus, a helper T cell epitope may be included in a hybrid antigen as the only epitope, and administered as an admixture with other hybrid antigens containing Class I epitope(s), or 5 the helper T cell epitope can be included in a tandem hybrid antigen as one of the epitopes. These are merely exemplary of the numerous variations upon the hybrid antigen compositions of the invention. 84 WO 2004/091493 PCT/US2004/010983 EXAMPLE 26 In a similar fashion to the previous example, the immunogenicity of a tandem hybrid antigen was evaluated with and without co-administration of a hybrid antigen containing the ovalbumin Class II peptide. 5 S250-72-12 Number of Spots per 300,000 cells Immunogen IAYFYPEL FAPGNYPAL Medium 19 Pg 9.3 = 4.7 17 9 0.7 L 0.6 NLLRLTGWFFRKIAYFYPELFFRKFAPGNYPAL 19 g NLLRLTGWFFRKIAYFYPELFFRKFAPGNYPAL Plus 44 5.1 58 5.2 0.7 0.6 20.8 gg NLLRLTGWFFRKTEWTSSNVMEERKIKV 250-72-15 Number of Spots per 300,000 Immunogen cells IAYFYPEL 25.5 g NLLRLTGWFFRKIAYFYPEL 3.7 3.1 25.5 g NLLRLTGWFFRKIAYFYPEL plus 133 & 11 34.7 jig NLLRLTGWFFRKTEWTSSNVMEERKIKV 25.5 ipg NLLRLTGWFFRKIAYFYPEL plus 2 5 M g 8 8 4 9 .9 25 Mg NLLRLTGWFFRKSIINFEKL 85 WO 2004/091493 PCT/US2004/010983 EXAMPLE 27 Similar experiments with hybrid antigens comprising a helper T cell epitope co administered with at least one tandem hybrid antigen, in the absence of co-administration of a heat shock protein, were also carried out. 5 250-72-12 Number of Spots per 300,000 cells Immunogen Medium SIINFEKL RGYVYQGL IAYFYPEL FAPGNYPAL 0.7 0.6 NT NT 9.3 ± 4.7 17 A 8.7 24 Vg NLLRLTGWFFRKIAYFYPELF FRKFAPGNYPAL 24 gg NLLRLTGWFFRKIAYFYPELF FRKFAPGNYPAL plus 0.7 ± 0.6 NT NT 44 ± 5.1 67 ± 5.5 21 gg NLLRLTGWFFRKTEWTSSNV MEERKIKV 15 pg N N 0.30.6 4.33.2 NLLRLTGWFFRKFAPGNYPA 00 NT NT 0.3 0.6 4.3 3.2 L 15 pg NLLRLTGWFFRKFAPGNYPA L plus 0 - 0 NT NT 2.3 2.1 58 5.2 21 lpg NLLRLTGWFFRKTEWTSSNV MEERKIKV 86 WO 2004/091493 PCT/US2004/010983 EXAMPLE 28 An immunization study using hybrid antigens containing human Class I (HLA-A2) epitopes was performed in IHIHD II mice as described above. Animals were immunized with a 5 complex made from 5 jig hsp70 and 33 Ag NLLRLTGWFFRKYMDGTMSQV. The ELISPOT results in cells per 300,000 were: Medium, 1.33 =- 0.58; splenocytes 1 ± 0; splenocytes plus YMDGTMSQV 123 :- 13; and splenocytes plus IMDQVPFSV 4 + 1. EXAMPLE 29 10 In another experiment using HHDII mice, an immunogenic HLA-A2 epitope from Trp-2 was used (SVYDFFVWL). Because this epitope is also a H2-Kb epitope, and the HHDII mice are on a B6 mouse (H2-Kb) background, an immune response induced against the Trp 2 peptide represents a breaking of tolerance to a self-epitope in the mouse model. The results of this experiment demonstrated that tolerance to this self-epitope was broken, and 15 the present invention is further directed to methods of breaking tolerance by administering the hybrid antigens and complexes of the invention. 87 WO 2004/091493 PCT/US2004/010983 xnaji-/-VU-/2-U3 Number of Spots per 300,000 cells Immunogen Medium SVYDFFVWL IMDQVPFSV Y'MDGTMSQV 4.33 jig NLLRLTGWFFRKSVYDFFVWL plus 0.5 ± 0.71 166 & 25 2.0 ± 1.4 3.5 -0.71 25 gg hsp70 8.66 ig NLLRLTGWFFRKSVYDFFVWL plus 3.5 0.71 114 - 11 7.7 + 2.1 11 4 3.1 25 jig hsp70 4.1 gg NLLRLTGWFFRKYMDGTMSQV plus 3.0 1.0 2.0 L 1.4 74 12.8 25 jig hsp70 4.1 ig NLLRLTGWFFRKIMDQVPQV plus 1.0 ± 1.4 2.0 ± 2.0 984 = 26 2.3 ± 1.5 25 pg hsp70 EXAMPLE 30 IHHDII mice were used to evaluate the immunogenicity of complexes of hsp70 and three 5 hybrid antigens comprising certain of the HIV viral component epitopes set forth in Example 27. 88 WO 2004/091493 PCT/US2004/010983 I-II)II-zVU- /2-0 / Immunogen Number of Spots per 300,000 cells Medium ILKEPVHGV VIYQYMDDL SLYNTVATL 36 gg NLLRLTGWFFR 1.0±1.0 34:112 0 0 NT KILKEPVHGV +25 pg hsp70 36 g NLLRLTGWFFR 0 ± 0 0.67 ± 0.58 24 1 6.1 NT KVIYQYMDDL + 25 tg hsp70 36 gg 0.67 4 NLLRLTGWFFR 0.67k NT NT 140 6.7 KSLYNTVATL 0.58 + 25 gg hsp70 NT=not tested EXAMPLE 31 5 Admixtures of hybrid antigens containing H2-Kb epitopes complexed with hsp70 were evaluated for immunogenicity in B6 mice as described above, 89 WO 2004/091493 PCT/US20041010983 Number of Spots per 300,000 cells Immunogen Medium Splenocytes SIIFEKL FAPGNVPAL IAVFYPEL 2 pg NLLRLTGWFFRKSIINFEKL 1 2 ±1 148 ±11 7:12 3± 2 + 13.7 ghsp70 10 pg NLLRLTGWFFRIIAYFYPEL~ o 2 ±2 3-11 8-12 47-+13 + 13.7 gg hsp70 10 Pg 3-3342 8 :66LI NLLRILTGWFFRKFAPGNYPAL 333328± ± + 13.7 pig hsp70 2 pg NLLRLTGWFPRKSIINFEKL + 10 Vg NLLRLTGWFFRK[AYFYPEL 2 4:k 2 94:6 4 9:1:3 29:h 4 27.4 pg hsp70 2 jig NLLRLTGWFFRXSIINFEKL + 10 ptg 33±0 61 5±74 NLLRLTGWFFRKFAPGNYPAL ±069 7 17 242 27.4 Vg hsp70 I0 gg NLLRLTGWFFRKIAYFYPEL + 3 34:3 4 3 46±h8 3912 10 ptg NLLRLTGWFFRKFAPGNYPAL + 27.4 pg hsp70______ 2pjg NLLRLTGWFFRKSLINFEKL + l0 pg 1 54±2 149±19 614:5 60 :b7 NLLRLTGWFFRKIAYFYPEL + 10 ptg NLLRLTGWFFRKFAPGNYPAL + 41 Lg hsp70 90 WO 2004/091493 PCT/US2004/010983 EXAMPLE 32 The immunogenicity of tandem hybrid antigens complexed with hsp70 was studied in B6 mice. S200-72-01 Number of Spots per 300,000 cells Immunogen Medium control Unpulsed SIINFEKL RGYVYQG control L 5.6 gg hsp70 + 033 0.41 0.67 ± 43 9.2 1±0 3 pg NLLRLTGWFFRKSIINFEKL 0.41 11.2 jig hsp70O+ 0.33 ± 11.2 gg hsp70 + 0.33 ± 0.41 0.334 1± 0.71 102 ±16 5.9 jig NLLRLTGWFFRKRGYVYQGL 0.41 11.2 pig hsp70 + 3 g NLLRLTGWFFRKSINFEKL + 0.67 0.82 1.7 ± 0.41 182 ± 11 113 4 10 5.9 gg NLLRLTGWFFRKRGYVYQGL 5.6 jig hsp70 + 0±0 411.4 456 ±19 113 ±1.1 4.8 gg NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 11.2 gg hsp70 + 0 ± 0 10 d 3.3 505± 57 90-11 9.6 gg NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 22.4 gg hsp70 + 0.67 - 0.82 1.7 1 0.41 289 : 26 130 -12 19.2 gg NLLRLTGWFFRKSIINFEKLFFRKRGYVYQGL 5.6 gg hsp70 + 0.33 1 0.41 2.3 ± 0.41 72 -9.5 98 4 9.2 4,8 [tg NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL 11.2 gghsp70 + 2 ± 0 2.3 ±1.5 370 16 617E 23 9.6 gg NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL 22.4 Ig hsp70 + 0.67 ± 0.41 4.0 - 2.1 336 ± 7.8 728 ± 12 19.3 gg NLLRLTGWFFRKRGYVYQGLFFRKSIINFEKL 5 91 WO 2004/091493 PCT/US2004/010983 111V present minvention is not to be limited in scope by the specific embodiments describe herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and the accompanying figures. Such modifications are intended to fall within the scope of the 5 appended claims. Various publications are cited herein, the contents of which are incorporated herein by reference in their entireties. 92

Claims (21)

1. A hybrid antigen comprising at least one antigenic domain of an infectious agent or tumor antigen, at least one binding domain that non-covalently binds to a heat shock protein, and at least one peptide linker there between selected from the group consisting of 5 Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO: 1003); Phe Arg (FR), Gln Leu Lys (QLK), Gln Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); and AAl-AA2-AA3-leucine, wherein AAl is A, S, V, E, G, L, or K, AA2 is K, V, or E; and AA3 is V, S, F, K, A, E, or 10 T.
2. A composition for inducing an immune response to an infectious agent or tumor antigen comprising at least one hybrid antigen of Claim 1.
3. A composition for inducing an immune response to an infectious agent or tumor antigen comprising a complex of at least one heat shock protein and at least one hybrid 15 antigen of Claim 1.
4. The composition of claim 3 wherein the heat shock protein is a hsp70.
5. A method for inducing an immune response to an infectious agent or tumor antigen comprising administering to a subject at least one hybrid antigen of Claim 1. 20
6. A method for inducing an immune response to an infectious agent or tumor antigen comprising administering to a subject a complex of: (a) a hybrid antigen of Claim 1; and (b) a heat shock protein; 25 wherein the hybrid antigen and the heat shock protein are non-covalently bound.
7. The method of claim 6 wherein the heat shock protein is a hsp70. 93 WO 2004/091493 PCT/US2004/010983
8. A method for treating an infectious disease or cancer comprising administering to a subject at least one hybrid antigen of Claim 1, wherein at least one antigenic domain is from the infectious disease or cancer. 5
9. A method for treating an infectious disease or cancer comprising administering to a subject a complex of: (a) a hybrid antigen of Claim 1, wherein at least one antigenic domain is from the infectious disease or cancer; and (b) a heat shock protein; 10 wherein the hybrid antigen and the heat shock protein are non-covalently bound.
10. The method of claim 9 wherein the heat shock protein is a hsp70.
11. A hybrid antigen consisting essentially of at least one antigenic domain of an 15 infectious agent or tumor antigen, at least one binding domain that non-covalently binds to a heat shock protein, and at least one peptide linker there between, and wherein peptide linker is selected from the group consisting of Phe Phe Arg Lys (FFRK; SEQ ID NO:1000); Phe Arg Lys (FRK); Phe Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gin 20 . Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); and AA1-AA2-AA3-leucine, wherein AA1 is A, S, V, E, G, L, or K, AA2 is K, V, or E; and AA3 is V, S, F, K, A, E, or T.
12. A composition for inducing an immune response to an infectious agent or tumor 25 antigen comprising at least one hybrid antigen of Claim 11.
13. A composition for inducing an immune response to an infectious agent or tumor antigen comprising a complex of at least one heat shock protein and at least one hybrid antigen of Claim 11.
14. The composition of claim 13 wherein the heat shock protein is a hsp70. 94 WO 2004/091493 PCT/US2004/010983
15. A method for inducing an immune response to an infectious agent or tumor antigen comprising administering to a subject at least one hybrid antigen of Claim 11. 5
16. A method for inducing an immune response to an infectious agent or tumor antigen comprising administering to a subject a complex of: (a) a hybrid antigen of Claim 11; and (b) a heat shock protein; wherein the hybrid antigen and the heat shock protein are non-covalently bound. 10
17. The method of claim 16 wherein the heat shock protein is a hsp70.
18. A method for treating an infectious disease or cancer comprising administering to a subject at least one hybrid antigen of Claim 11, wherein at least one antigenic domain is 15 from the infectious disease or cancer.
19. A method for treating an infectious disease or cancer comprising administering to a subject a complex of: (a) a hybrid antigen of Claim 1, wherein the antigenic domain is from the 20 infectious disease or cancer; and (b) a heat shock protein; wherein the hybrid antigen and the heat shock protein are non-covalently bound.
20. The method of claim 19 wherein the heat shock protein is a hsp70. 25
21. A peptide that is Phe Phe Arg Lys (FFRK; SEQ ID NO: 1000); Phe Arg Lys (FRK); Phe.Arg Lys Asn (FRKN, SEQ ID NO: 1002); Arg Lys Asn (RKN); Phe Phe Arg Lys Asn (FFRKN, SEQ ID NO:1003); Phe Arg (FR), Gln Leu Lys (QLK), Gin Leu Glu (QLE), Ala Lys Val Leu (AKVL; SEQ ID NO:1001); Lys Asn (KN); Arg Lys (RK); or AAl-AA2 95 WO 2004/091493 PCT/US2004/010983 AA3-leucine, wherein AA1 is A, S, V, E, G, L, or K, AA2 is K, V, or E; and AA3 is V, S, F, K, A, E, or T. 96
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