Methods and composition related to Th-1 dendritic cells
Certain embodiments of the invention are directed to methods for inducing an immunologic response to a tumor in a patient using mature dendritic cells transfected with a nucleic acid composition encoding one or more tumor antigens and loaded with a corresponding tumor antigen composition.
1. A method for inducing an immunologic response to a tumor in a patient comprising:
(a) obtaining monocytic dendritic cell precursors from the patient, wherein the patient has breast cancer, a glioma, melanoma, pancreatic cancer or prostate cancer;
(b) culturing the monocytic dendritic cell precursors to induce differentiation into immature dendritic cells;
(c) differentiating the immature dendritic cells into mature dendritic cells by providing to said cells a tumor antigen composition comprising at least one Major Histocompatibility Complex (MHC) Class I epitope and at least one MHC Class II epitope, wherein the Class I and the Class II epitopes have a sequence overlap of at least 5 amino acids;
(d) administering the mature dendritic cells to the patient.
2. The method of claim 1 , wherein the mature dendritic cells are selected for CD83 expression, wherein the selected mature dendritic cells are enriched for cells expressing increased levels of CD83 as compared to reference dendritic cells contacted with a tumor antigen composition and not a nucleic acid composition.
3. The method of claim 2 , wherein CD83 expression is at least 10-40% higher than the reference dendritic cells.
4. The method of claim 1 , further comprising screening a patient or patient tumor for expression of one or more tumor antigens.
5. The method of claim 1 , wherein the patient has breast cancer.
6. The method of claim 1 , wherein the patient has a glioma.
7. The method of claim 1 , wherein the patient has melanoma.
8. The method of claim 1 , wherein the patient has pancreatic cancer.
9. The method of claim 1 , wherein the patient has prostate cancer.
10. The method of claim 1 , wherein differentiating the immature dendritic cells into mature dendritic cells in step (c) comprises:
(i) transfecting into the immature dendritic cells a nucleic acid composition encoding one or more tumor antigens; and
(ii) contacting the immature dendritic cells with a tumor antigen composition.
11. The method of claim 10 , wherein the nucleic acid composition comprises total nucleic acid from a tumor source.
12. The method of claim 10 , wherein the nucleic acid composition comprises mRNA isolated from a tumor source.
13. The method of claim 12 , wherein the isolated mRNA is enriched for mRNA encoding tumor specific antigens.
14. The method of claim 13 , wherein the isolated mRNA is subjected to mRNA subtraction using non-tumor cell RNA.
15. The method of claim 10 , wherein the tumor antigen composition is an enriched tumor antigen composition.
16. The method of claim 15 , wherein the enriched tumor antigen composition comprises a cellular fraction from cells of a tumor source.
17. The method of claim 15 , wherein the cells from the tumor source are selected by removing cells expressing proteins that are typically not expressed or expressed at significantly reduced levels in a tumor cell.
18. The method of claim 15 , wherein the cells from the tumor source are selected using cell surface markers preferentially expressed by tumor cells.
19. The method of claim 15 , wherein the enriched tumor antigen composition comprises one or more protein fractions of a tumor cell lysate.
20. The method of claim 19 , wherein the enriched tumor antigen composition is produced by contacting a tumor cell lysate with a protein array that preferentially binds non-tumor specific antigens.
21. The method of claim 10 , wherein the nucleic acid composition comprises an expression construct.
22. The method of claim 21 , wherein the expression construct encodes one or more tumor antigen.
23. The method of claim 10 , wherein the immature dendritic cell is contacted with a tumor antigen composition comprising a recombinant identified tumor antigen.
24. The method of claim 10 , wherein the nucleic acid composition encodes components of the tumor antigen composition.
25. The method of claim 10 , wherein the immature dendritic cells are transfected with the nucleic acid composition prior to contact with the tumor antigen composition.
26. The method of claim 10 , wherein the immature dendritic cells are contacted with the tumor antigen composition prior to transfection with the nucleic acid composition.
27. The method of claim 10 , wherein the immature dendritic cells are simultaneously transfected with the nucleic acid composition and contacted with the tumor antigen composition.
28. The method of claim 1 , wherein the MHC Class I or MHC Class II epitopes are comprised on separate peptides.
29. The method of claim 28 , wherein the peptides are synthetic peptides.