Polypeptides and polynucleotides for enhancing immune reactivity to HER-2 protein
Compositions for stimulating the immune system and for treating malignancies associated with overexpression of the HER-2 protein are provided. Such compositions include immunogenic epitopes of the HER-2 proteins and chimeric and multivalent peptides which comprise such epitopes. The present invention also relates to polynucleotides which encode the chimeric peptides. Also provided are pharmaceutical compositions comprising such immunogenic compositions. Methods for stimulating an immune response to HER-2 protein are provided. Methods for treating breast cancer, ovarian cancer, prostate cancer, colon cancer and lung cancer are provided.
1. A chimeric peptide composition for stimulating an immune response to HER-2 protein, wherein said chimeric peptide comprises: a HER-2 B cell epitope, a T helper (Th) epitope, and a linker joining said HER-2 B cell epitope to said Th epitope, wherein:
the HER-2 B cell epitope consists of KPDLSYMPIWKFPDEEGA, SEQ ID NO: 11;
the Th epitope comprises a sequence selected from the group consisting of:
NSVDDALINSTIYSYFPSV, SEQ ID NO: 13;
PGINGKAIHLVNNQSSE, SEQ ID NO: 14;
QYIKANSKFIGITEL, SEQ ID NO: 15;
FNNFTVSFWLRVPKVSASHLE, SEQ ID NO: 16;
LSEIKGVIVHRLEGV, SEQ ID NO: 17;
FFLLTRILTIPQSLN, SEQ ID NO: 18; and
TCGVGVRVRSRVNAANKKPE, SEQ ID NO: 19; and
the linker is from 1 to 15 amino acids in length.
2. The chimeric peptide of claim 1 wherein said linker is from 2 to 15 amino acids in length.
3. The chimeric peptide of claim 1 wherein the Th epitope comprises LSEIKGVIVHRLEGV, SEQ ID NO: 17.
4. The chimeric peptide of claim 1 wherein the linker comprises the sequence GPSL, SEQ ID NO: 20.
5. The chimeric peptide of claim 1 further comprising a second HER-2 B cell epitope comprising an amino acid sequence selected from the group consisting of:
TGTDMKLRLPASPETHLDM, SEQ ID NO: 1:
AVLDNGDPLNNTTPVTGASPGG, SEQ ID NO: 2:
LWKDIFHKNNQLALTLIDTNRS, SEQ ID NO: 3:
TLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLT, SEQ ID NO: 4:
ALVTYNTDTFESMPNPEGRYT, SEQ ID NO: 5:
PLHNQEVTAEDGTQRAEKCSKPCA, SEQ ID NO: 6:
PESFDGDPASNTAPLQPE, SEQ ID NO: 7:
LYISAWPDSLPDLSVFQNLQ, SEQ ID NO: 8:
LFRNPHQALLHTANRPEDE, SEQ ID NO: 9:
CLPCHPECQPQNGSVTCFGPEADQCVACAHYKDP, SEQ ID NO: 10:
KPDLSYMPIWKFPDEEGA, SEQ ID NO: 11: and
INGTHSCVDLDDKGCPAEQRAS, SEQ ID NO: 12.
6. A method of stimulating an immune response in a subject comprising administering a pharmaceutical composition to said subject, said pharmaceutical composition comprising:
the chimeric peptide of claim 1 , and
a pharmaceutically acceptable vehicle.
7. The method of claim 6 wherein the subject is a human and has one of the following cancers or a predisposition to one of the following cancers: breast cancer, ovarian cancer, lung cancer, prostate cancer, and colon cancer.
8. The method of claim 6 wherein the vehicle is biodegradable and is selected from the group consisting of an emulsion comprising a pharmaceutically acceptable oil/water emulsion and a biodegradable microsphere or nanosphere comprising a polylactide-polyglycolic acid polymer.
9. The method of claim 8 wherein the oil is squalene or squalane.
10. The method of claim 8 wherein the microsphere is from 0.1 to 50 nanometers in diameter and comprises poly(D,L lactide-co-glycide).
11. The chimeric peptide of claim 5 , further comprising a second linker joining the first HER-2 B cell epitope to the second HER-2 B cell epitope.
12. The chimeric peptide of claim 11 , wherein the second linker is from 1 to 15 amino acids in length.
13. The chimeric peptide of claim 12 , wherein the second linker comprises the sequence GPSL, SEQ ID NO: 20.
14. The chimeric peptide of claim 1 wherein the Th epitope comprises NSVDDALINSTIYSYFPSV, SEQ ID NO: 13.
15. The chimeric peptide of claim 1 wherein the Th epitope comprises PGINGKAIHLVNNQSSE, SEQ ID NO: 14.
16. The chimeric peptide of claim 1 wherein the Th epitope comprises QYIKANSKFIGITEL, SEQ ID NO: 15.
17. The chimeric peptide of claim 1 wherein the Th epitope comprises FNNFTVSFWLRVPKVSASHLE, SEQ ID NO: 16.
18. The chimeric peptide of claim 1 wherein the Th epitope comprises FFLLTRILTIPQSLN, SEQ ID NO: 18.
19. The chimeric peptide of claim 1 wherein the Th epitope comprises TCGVGVRVRSRVNAANKKPE, SEQ ID NO: 19.