Enhancing the T-cell stimulatory capacity of human antigen presenting cells in vitro and in vivo and its use in vaccination
We provide new methods of in vitro or in vivo enhancing the T-cell stimulatory capacity of human DCs and the use thereof in cancer vaccination. The method includes the introduction of different molecular adjuvants to human DCs by contacting or modifying them with mRNA or DNA molecule(s) encoding CD40L, and CD70 or constitutively active TLR4 (caTLR4).
1. A method for the treatment of cancer, comprising the step of intratumoral administering to a subject in need thereof one or more isolated mRNA molecule(s) encoding for:
a functional immunostimulatory protein CD40L, constitutively active TLR4 (caTLR4), and a functional immunostimulatory protein CD70; wherein said method does not include the administration of dendritic cells to said subject.
2. The method according to claim 1 , additionally comprising the step of administering target-specific antigen(s) before, after, or simultaneously with the one or more isolated mRNA molecule(s).
3. The method of claim 2 , wherein the target-specific antigen is a tumor antigen.
4. The method of claim 2 , wherein said target-specific antigen(s) is selected from the group consisting of: total mRNA isolated from (a) target cell(s), one or more specific target mRNA molecules, protein lysates of (a) target cell(s), specific proteins from (a) target cell(s), a synthetic target-specific peptide or protein, and synthetic mRNA or DNA encoding a target-specific antigen or its derived peptide(s).
5. The method according to claim 1 , wherein prior to said intratumoral administering, the tumor was treated by ablation, thereby freeing antigens.
6. The method of claim 1 , additionally comprising the administration of mRNA or DNA encoding one or more of the following molecules: IL-12p70, EL-selectin, CCR7, and/or 4-1BBL.
7. The method according to claim 1 , wherein said one or more isolated mRNA molecule(s) encoding the immunostimulatory proteins are part of a single mRNA molecule.
8. The method according to claim 7 , wherein the single mRNA molecule is capable of expressing the two or more proteins simultaneously.
9. The method according to claim 1 , wherein said mRNA is protected.
10. The method according to claim 1 , wherein the cancer is chosen from leukemia, non-small cell lung cancer, small cell lung cancer, CNS cancer, melanoma, ovarian cancer, kidney cancer, prostate cancer, breast cancer, glioma, colon cancer, bladder cancer, sarcoma, pancreatic cancer, colorectal cancer, head and neck cancer, liver cancer, bone cancer, bone marrow cancer, stomach cancer, duodenum cancer, oesophageal cancer, thyroid cancer, hematological cancer, and lymphoma.
11. The method according to claim 1 , wherein the method does not include the administration of a target specific antigen to said subject.