Hybrid antigens comprising an antigenic domain and improved heat shock protein binding domains are described which are useful for the induction of an immune response to the antigenic domain and thus can be used to treat infectious diseases and cancers that express an antigen of the antigenic domain.

Description of the Invention

SUMMARY OF THE INVENTION

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 complex with a heat shock protein is administered to the subject. The hybrid antigen comprises an antigenic domain and a heat shock protein binding domain. 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 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. In specific, non-limiting embodiments of the invention, the heat shock protein binding domain comprises a peptide having the sequence: Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), or Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419).

In alternate embodiments, the heat shock protein binding domain comprises a peptide have a sequence among SEQ ID NOs:132-185 and 193-356.

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 shock protein, in an effective amount to a subject in need of such treatment.

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.

Thus, in a first aspect, the invention is directed to a hybrid antigen consisting 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 binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356.

In a second aspect, the invention is directed to a hybrid antigen consisting essentially of a plurality of antigenic domains of an infectious agent or tumor antigen, a binding domain that non-covalently binds to a heat shock protein, and peptide linkers separating the antigenic domains and the binding domain, and wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In a particular embodiment, at least one of the antigenic domains in the aforementioned hybrid antigen is a T helper epitope.

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 wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In a particular embodiment, the aforementioned hybrid antigen has a peptide linker separating the antigenic domain and the binding domain.

In a fourth aspect, the invention is directed to a hybrid antigen comprising a plurality of antigenic domains of an infectious agent or tumor antigen and a binding domain that non-covalently binds to a heat shock protein, and wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In a particular embodiment, peptide linkers separate the antigenic domains and the binding domain. In yet another embodiment, at least one of the antigenic domains is a T helper epitope.

In a fifth 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 comprising an antigenic domain of the infectious agent or tumor antigen and a binding domain that non-covalently binds to a heat shock protein, and wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In one embodiment, a peptide linker separates the antigenic domain and the binding domain. In another embodiment, the composition comprises a plurality of hybrid antigens, and one of the hybrid antigens can comprise a T helper epitope.

In a sixth 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 comprising a plurality of antigenic domains at least one of which is from the infectious agent or tumor antigen, and a binding domain that non-covalently binds to a heat shock protein, wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In one embodiment, peptide linkers separate the antigenic domains from the binding domain. In another embodiment, at least one of the antigenic domains comprises a T helper epitope.

In a seventh 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 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 binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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.

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 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 binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In one embodiment, at least one of the antigenic domains comprises a T helper epitope.

In a ninth 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) a hybrid antigen comprising at least one antigenic domain of the infectious agent or tumor antigen, and a binding domain comprising a peptide that non-covalently binds to a heat shock protein; and (b) a heat shock protein;

wherein the hybrid antigen and the heat shock protein are non-covalently bound, and wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In one embodiment, the complex comprises a plurality of hybrid antigens. In an 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, 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 an embodiment of this aspect of the invention, a peptide linker separates the antigenic domain and the binding domain. In another embodiment of this aspect of the invention, the heat shock protein is a hsp70.

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 separating the antigenic domain and the binding domain, and wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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 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.

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, and a binding domain comprising a peptide that non-covalently binds to a heat shock protein, wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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 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 antigenic domains. In another embodiment of this aspect of the invention, a peptide linker separates the antigenic domain and the binding domain.

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-covalently binds to a heat shock protein, and a peptide linker separating the antigenic domain and the binding domain, and wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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 further embodiment, the complex comprises a plurality of hybrid antigens, at least one of the hybrid antigens comprising a plurality of antigenic domains.

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) a hybrid antigen comprising at least one antigenic domain of an infectious agent or tumor antigen associated with the infectious disease or cancer, and a binding domain comprising a peptide that non-covalently binds to a heat shock protein; and (b) a heat shock protein;

wherein the hybrid antigen and the heat shock protein are non-covalently bound, and wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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 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.

In a fourteenth 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, 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, 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 binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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. In yet another aspect, the hybrid antigen comprises a plurality of antigenic domains. In yet another aspect, at least one of the antigenic domains 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.

In a fifteen 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 comprising at least one antigenic domain of an infectious agent or tumor antigen associated with the infectious disease or cancer, and a binding domain comprising a peptide that non-covalently binds to a heat shock protein, wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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. 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 domain.

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 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 binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. 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 antigens comprising a plurality of antigenic domains.

In a seventeenth aspect, the invention is directed to a peptide comprising Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356.

In an eighteenth aspect, the invention is directed to an immunogenic polypeptide comprising a plurality of antigenic domains, and a binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356.

In a nineteenth aspect, the invention is directed to the peptides Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356.

In a twentieth aspect, the invention is directed to a polynucleotide encoding any of the aforementioned hybrid antigens.

In a twenty-first 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 agent or tumor antigen and a heat shock protein binding domain.

In a twenty-second 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 is a hsp70.

In a twenty-third aspect, the invention is directed to polynucleotides encoding Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Tyr Trp (SEQ ID NO:418), Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419), or any of SEQ ID NOs:132-185 and 193-356. In a further embodiment, the invention is directed to polynucleotides encoding hybrid antigens as described above. In another embodiment, the invention is directed to inducing an immune response to an infectious agent or cancer comprising administering to a subject a polynucleotide encoding a hybrid antigen as mentioned above, optionally together with a polynucleotide encoding a heat shock protein, preferably hsp70. In a further embodiment, the invention is directed to treating an infectious disease or cancer comprising administering to a subject a polynucleotide encoding a hybrid antigen as mentioned above, optionally together with a polynucleotide encoding a heat shock protein, preferably hsp70.

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 leukemia, or a carcinoma, a melanoma, carcinoma of the breast, carcinoma of the prostate, 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 antigen of the infectious agent or cancer antigen, respectively, is useful for treating the corresponding infectious disease or cancer.

This application claims priority under 35 U.S.C. .sctn. 119(e) to provisional applications 60/447,142, filed Feb. 13, 2003; 60/462,469, filed Apr. 11, 2003; 60/463,746, filed Apr. 18, 2003; and 60/503,417, filed Sep. 16, 2003, all four of which are incorporated herein by reference in their entireties.
 

Claim 1 of 45 Claims

1. A hybrid antigen comprising at least one antigenic domain of an infectious agent or tumor antigen and a binding domain that non-covalently binds to a heat shock protein, wherein the binding domain comprises Asn Leu Leu Arg Leu Thr Gly Trp (SEQ ID NO:417), Phe Tyr Gln Leu Ala Leu Thr Trp (SEQ ID NO:186), or Arg Lys Leu Phe Phe Asn Leu Arg Trp (SEQ ID NO:419).

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