This invention provides methods of treating and vaccinating against an antigen-expressing tumor and inducing an immune response against a sub-dominant epitope of antigen, comprising a fusion of an LLO fragment to the antigen or a recombinant Listeria strain expressing the antigen. The present invention also provides recombinant peptides comprising a listeriolysin (LLO) protein fragment fused to a Her-2 protein or fragment thereof, recombinant Listeria strains expressing a Her-2 protein, vaccines and immunogenic compositions comprising same, and methods of inducing an anti-Her-2 immune response and treating and vaccinating against a Her-2-expressing tumor, comprising same.
1. A recombinant form of Listeria comprising a nucleotide molecule encoding a recombinant polypeptide comprising an N-terminal fragment of a listeriolysin (LLO) protein fused to a fragment of a Her-2 protein, wherein said fragment of a Her-2 protein consists of amino acids 303-501, 479-655, or 1020-1255 of SEQ ID NO: 40 or amino acids 303-501, 479-652, or 1020-1255 of SEQ ID NO: 43.
2. The recombinant form of Listeria of claim 1 , wherein said N-terminal fragment of a listeriolysin (LLO) protein is non-hemolytic.
3. An immunogenic composition comprising the recombinant form of Listeria of claim 1 .
4. A recombinant form of Listeria comprising a nucleotide molecule encoding a fragment of a Her-2 protein wherein said fragment of a Her-2 protein consists of amino acids 303-501, 479-655, or 1020-1255 of SEQ ID NO: 40 or amino acids 303-501, 479-652, or 1020-1255 of SEQ ID NO: 43.
5. The recombinant form of Listeria of claim 4 , wherein said Her-2 protein is a human Her-2 protein.
6. An immunogenic composition comprising the recombinant form of Listeria of claim 4 .
7. A recombinant L. monocytogenes strain expressing a recombinant fusion polypeptide comprising an N-terminal listeriolysin (LLO) fragment fused to a fragment of a Her-2 protein, wherein said fragment consists of amino acids 303-501 or 479-655 of SEQ ID NO: 40 or amino acids 303-501 or 479-652 of SEQ ID NO: 43.
8. A recombinant L. monocytogenes strain expressing a recombinant fusion polypeptide comprising an N-terminal listeriolysin (LLO) fragment fused to a fragment of a Her-2 protein, wherein said fragment consists of amino acids 1020-1255 of SEQ ID NO: 40 or amino acids 1020-1255 of SEQ ID NO: 43.
9. The recombinant form of Listeria of claim 1 , wherein said fragment of a Her-2 protein consists of amino acids 303-501 of SEQ ID NO: 40.
10. The recombinant form of Listeria of claim 1 , wherein said fragment of a Her-2 protein consists of amino acids 303-501 of SEQ ID NO: 43.
11. The recombinant form of Listeria of claim 1 , wherein said fragment of a Her-2 protein consists of amino acids 479-655 of SEQ ID NO: 40.
12. The recombinant form of Listeria of claim 1 , wherein said fragment of a Her-2 protein consists of amino acids 479-652 of SEQ ID NO: 43.
13. The recombinant form of Listeria of claim 1 , wherein said fragment of a Her-2 protein consists of amino acids 1020-1255 of SEQ ID NO: 40.
14. The recombinant form of Listeria of claim 1 , wherein said fragment of a Her-2 protein consists of amino acids 1020-1255 of SEQ ID NO: 43.
15. The recombinant form of Listeria of claim 4 , wherein said fragment of a Her-2 protein consists of amino acids 303-501 of SEQ ID NO: 40.
16. The recombinant form of Listeria of claim 4 , wherein said fragment of a Her-2 protein consists of amino acids 303-501 of SEQ ID NO: 43.
17. The recombinant form of Listeria of claim 4 , wherein said fragment of a Her-2 protein consists of amino acids 479-655 of SEQ ID NO: 40.
18. The recombinant form of Listeria of claim 4 , wherein said fragment of a Her-2 protein consists of amino acids 479-652 of SEQ ID NO: 43.
19. The recombinant form of Listeria of claim 4 , wherein said fragment of a Her-2 protein consists of amino acids 1020-1255 of SEQ ID NO: 40.
20. The recombinant form of Listeria of claim 4 , wherein said fragment of a Her-2 protein consists of amino acids 1020-1255 of SEQ ID NO: 43.
21. A method of inducing an anti-Her-2 immune response in a subject, comprising administering to said subject the recombinant form of Listeria of claim 1 , thereby inducing an anti-Her-2 immune response in said subject.
22. The method of claim 21 , wherein said immune response comprises a CD8+ T cell-mediated response.
23. The method of claim 21 , whereby said immune response comprises an immune response to a subdominant epitope of said Her-2 protein.
24. A method of impeding the growth of a Her-2-expressing tumor in a subject, comprising administering to said subject the recombinant form of Listeria of claim 1 , whereby said subject mounts an immune response against said Her-2-expressing tumor, thereby impeding the growth of said Her-2-expressing tumor in said subject.
25. The method of claim 24 , wherein said immune response comprises a CD8+ T cell-mediated response.
26. The method of claim 24 , whereby said immune response against said Her-2-expressing tumor comprises an immune response to a subdominant epitope of said Her-2 protein.
27. A method of shrinking a Her-2-expressing tumor in a subject, comprising administering to said subject the recombinant form of Listeria of claim 1 , whereby said subject mounts an immune response against said Her-2-expressing tumor, thereby shrinking said Her-2-expressing tumor in said subject.
28. The method of claim 27 , wherein said immune response comprises a CD8 + T cell-mediated response.
29. The method of claim 27 , whereby said immune response against said Her-2-expressing tumor comprises an immune response to a subdominant epitope of said Her-2 protein.
30. A method of breaking an immune tolerance of a subject to an antigen-expressing tumor, comprising administering to said subject the recombinant form of Listeria of claim 1 , wherein said Her-2 protein has one or more dominant CD8 + T cell epitopes and wherein said fragment does not contain said any of said dominant CD8 + T cell epitopes, whereby said subject mounts an immune response against said antigen-expressing tumor, thereby breaking an immune tolerance of a subject to an antigen-expressing tumor.
31. The method of claim 30 , wherein said Her-2 protein is expressed at a detectable level on a non-tumor cell of said subject.
32. The method of claim 30 , wherein at least part of said immune response is a CD8 + T cell response directed against an epitope of said Her-2 protein that is not any of said dominant CD8 + T cell epitopes.
33. A method of inducing a CD8 + T cell-mediated immune response in a subject against a subdominant CD8 + T cell epitope of an antigen, comprising administering to said subject the recombinant form of Listeria of claim 1 , thereby inducing a CD8 + T cell-mediated immune response against a subdominant CD8 + T cell epitope of an antigen.
34. A method of inducing an anti-Her-2 immune response in a subject, comprising administering to said subject the recombinant form of Listeria of claim 4 , thereby inducing an anti-Her-2 immune response in said subject.
35. The method of claim 34 , wherein said immune response comprises a CD8+ T cell-mediated response.
36. The method of claim 34 , whereby said immune response comprises an immune response to a subdominant epitope of said Her-2 protein.
37. A method of impeding the growth of a Her-2-expressing tumor in a subject, comprising administering to said subject the recombinant form of Listeria of claim 4 , whereby said subject mounts an immune response against said Her-2-expressing tumor, thereby impeding the growth of said Her-2-expressing tumor in said subject.
38. The method of claim 37 , wherein said immune response comprises a CD8+ T cell-mediated response.
39. The method of claim 37 , whereby said immune response against said Her-2-expressing tumor comprises an immune response to a subdominant epitope of said Her-2 protein.
40. A method of shrinking a Her-2-expressing tumor in a subject, comprising administering to said subject the recombinant form of Listeria of claim 4 , whereby said subject mounts an immune response against said Her-2-expressing tumor, thereby shrinking said Her-2-expressing tumor in said subject.
41. The method of claim 40 , wherein said immune response comprises a CD8 + T cell-mediated response.
42. The method of claim 40 , whereby said immune response against said Her-2-expressing tumor comprises an immune response to a subdominant epitope of said Her-2 protein.
43. A method of breaking an immune tolerance of a subject to an antigen-expressing tumor, comprising administering to said subject the recombinant form of Listeria of claim 4 , wherein said Her-2 protein has one or more dominant CD8 + T cell epitopes and wherein said fragment does not contain said any of said dominant CD8 + T cell epitopes, whereby said subject mounts an immune response against said antigen-expressing tumor, thereby breaking an immune tolerance of a subject to an antigen-expressing tumor.
44. The method of claim 43 , wherein said Her-2 protein is expressed at a detectable level on a non-tumor cell of said subject.
45. The method of claim 43 , wherein at least part of said immune response is a CD8 + T cell response directed against an epitope of said Her-2 protein that is not any of said dominant CD8 + T cell epitopes.
46. A method of inducing a CD8 + T cell-mediated immune response in a subject against a subdominant CD8 + T cell epitope of an antigen, comprising administering to said subject the recombinant form of Listeria of claim 4 , thereby inducing a CD8 + T cell-mediated immune response against a subdominant CD8 + T cell epitope of an antigen.
47. A method of inducing an anti-Her-2 immune response in a subject, comprising administering to said subject the recombinant form of Listeria of claim 7 , thereby inducing an anti-Her-2 immune response in said subject.
48. The method of claim 47 , wherein said immune response comprises a CD8+ T cell-mediated response.
49. The method of claim 47 , whereby said immune response comprises an immune response to a subdominant epitope of said Her-2 protein.
50. A method of impeding the growth of a Her-2-expressing tumor in a subject, comprising administering to said subject the recombinant form of Listeria of claim 7 , whereby said subject mounts an immune response against said Her-2-expressing tumor, thereby impeding the growth of said Her-2-expressing tumor in said subject.
51. The method of claim 50 , wherein said immune response comprises a CD8+ T cell-mediated response.
52. The method of claim 50 , whereby said immune response against said Her-2-expressing tumor comprises an immune response to a subdominant epitope of said Her-2 protein.
53. A method of shrinking a Her-2-expressing tumor in a subject, comprising administering to said subject the recombinant form of Listeria of claim 7 , whereby said subject mounts an immune response against said Her-2-expressing tumor, thereby shrinking said Her-2-expressing tumor in said subject.
54. The method of claim 53 , wherein said immune response comprises a CD8 + T cell-mediated response.
55. The method of claim 53 , whereby said immune response against said Her-2-expressing tumor comprises an immune response to a subdominant epitope of said Her-2 protein.
56. A method of breaking an immune tolerance of a subject to an antigen-expressing tumor, comprising administering to said subject the recombinant form of Listeria of claim 7 , wherein said Her-2 protein has one or more dominant CD8 + T cell epitopes and wherein said fragment does not contain said any of said dominant CD8 + T cell epitopes, whereby said subject mounts an immune response against said antigen-expressing tumor, thereby breaking an immune tolerance of a subject to an antigen-expressing tumor.
57. The method of claim 56 , wherein said Her-2 protein is expressed at a detectable level on a non-tumor cell of said subject.
58. The method of claim 56 , wherein at least part of said immune response is a CD8 + T cell response directed against an epitope of said Her-2 protein that is not any of said dominant CD8 + T cell epitopes.
59. A method of inducing a CD8 + T cell-mediated immune response in a subject against a subdominant CD8 + T cell epitope of an antigen, comprising administering to said subject the recombinant form of Listeria of claim 7 , thereby inducing a CD8 + T cell-mediated immune response against a subdominant CD8 + T cell epitope of an antigen.
60. A recombinant amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 36.
61. A recombinant amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 37.
62. A recombinant amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 39.