초록 ▼

Modified antigen presenting cells provided herein have improved lifespan and immunogenicity compared to unmodified antigen presenting cells, and are useful for immunotherapy. The modified antigen presenting cells express an altered protein kinase, referred to herein as “Akt.” The altered Akt associa

Modified antigen presenting cells provided herein have improved lifespan and immunogenicity compared to unmodified antigen presenting cells, and are useful for immunotherapy. The modified antigen presenting cells express an altered protein kinase, referred to herein as “Akt.” The altered Akt associates with the cell membrane with greater frequency than unaltered Akt, and is referred to herein as “membrane-targeted Akt.”

대표청구항 ▼

1. A method for preparing a modified dendritic cell, which comprises: transferring a polynucleotide into a dendritic cell, wherein the polynucleotide encodes a membrane-targeted Akt protein comprising a dual acylation region and a mammalian Akt region, wherein the mammalian Akt region comprises the

1. A method for preparing a modified dendritic cell, which comprises: transferring a polynucleotide into a dendritic cell, wherein the polynucleotide encodes a membrane-targeted Akt protein comprising a dual acylation region and a mammalian Akt region, wherein the mammalian Akt region comprises the amino acid sequence of SEQ ID NO: 6, wherein the membrane-targeted Akt protein is expressed in the dendritic cell and the modified dendritic cell survives longer than dendritic cells that do not express the protein. 2. A method for preparing a modified dendritic cell, which comprises: transferring a polynucleotide into a dendritic cell, wherein the polynucleotide encodes a membrane-targeted Akt protein comprising a dual acylation region and a mammalian Akt region, wherein the mammalian Akt region consists of the amino acid sequence of SEQ ID NO: 6, wherein the membrane-targeted Akt protein is expressed in the dendritic cell and the modified dendritic cell survives longer than dendritic cells that do not express the protein. 3. The method of claim 1, wherein the acylation region comprises the amino acid sequence of SEQ ID NO: 8. 4. The method of claim 1, wherein the acylation region consists of the amino acid sequence of SEQ ID NO: 8. 5. The method of claim 1, wherein the polynucleotide is from a virus. 6. The method of claim 5, wherein the dendritic cell is contacted with a virus that contains the polynucleotide. 7. The method of claim 1, wherein the polynucleotide comprises a constitutively active promoter operably linked to the polynucleotide that encodes the membrane-targeted Akt protein. 8. The method of claim 1, wherein the polynucleotide is transferred into the dendritic cell by a method selected from the group consisting of calcium phosphate precipitation, electroporation, direct microinjection, liposome transfer, gene bombardment, receptor-mediated transfection, receptor-mediated endocytosis, and viral transfer. 9. The method of claim 1, wherein the polynucleotide is transferred into the dendritic cell using an adenoviral vector. 10. The method of claim 1, wherein the dual acylation region is a Fyn myristoylation sequence. 11. The method of claim 2, wherein the acylation region comprises the amino acid sequence of SEQ ID NO: 8. 12. The method of claim 2, wherein the acylation region consists of the amino acid sequence of SEQ ID NO: 8. 13. The method of claim 12, wherein the modified dendritic cell is loaded with antigen and wherein the modified dendritic cell presents a greater amount of the antigen than dendritic cells that do not include the polynucleotide. 14. The method of claim 12, wherein the modified dendritic cell is more immunogenic than dendritic cells that do not include the polynucleotide. 15. The method of claim 12, which comprises contacting the modified dendritic cell with an antigen. 16. The method of claim 15, wherein the antigen is prostate specific membrane antigen. 17. The method of claim 16, wherein the antigen has a sequence of SEQ ID NO: 10, or an immunogenic fragment thereof. 18. The method of claim 12, wherein the dendritic cell is a human cell. 19. The method of claim 1, wherein the modified dendritic cell is loaded with antigen and wherein the modified dendritic cell presents a greater amount of the antigen than dendritic cells that do not include the polynucleotide. 20. The method of claim 1, wherein the modified dendritic cell is more immunogenic than dendritic cells that do not include the polynucleotide. 21. The method of claim 1, which comprises contacting the modified dendritic cell with an antigen. 22. The method of claim 19, wherein the antigen is prostate specific membrane antigen. 23. The method of claim 20, wherein the antigen comprises a sequence of SEQ ID NO: 10, or an immunogenic fragment thereof. 24. The method of claim 1, wherein the dendritic cell is a human cell.