Generation of antigen specific T cells
The present invention is directed to a method of generating antigen specific T cells. Furthermore, the invention is directed to antigen specific T cells, isolated transgenic TCR's, pharmaceutical compositions containing same and their use in adoptive cell therapy. This invention in particular pertains to the use of cells co-expressing allogeneic MHC molecules and antigens to induce peptide-specific T cells from non-selected allogeneic T cell repertoires.
1. A method of generating antigen specific T cells, comprising:
a) providing a nucleic acid encoding a patient-derived MHC molecule; and an antigen or a second nucleic acid encoding said antigen;
b) co-transfecting or introducing both compounds as defined in a) into antigen presenting cells (APCs) derived from a healthy donor who is allogeneic to the patient;
c) priming peripheral blood lymphocytes (PBLs) derived from the healthy donor with said APCs; and
d) selecting antigen-specific T cells which are specific for an MHC-antigen ligand that comprises the patient-derived MHC molecule bound to the antigen.
2. The method of claim 1 , wherein the APCs are selected from the group consisting of dendritic cells, activated B cells, monocytes, macrophages, and activated T cells.
3. The method of claim 1 , wherein the MHC molecule and the antigen are present as a mixture of antigen and nucleic acid encoding the MHC molecule.
4. The method of claim 1 , wherein the antigen is selected from the group consisting of pathogenic agents derived from viruses, bacteria, protozoa, parasites, tumor cells, tumor cell associated antigens, autoantigens and functional parts thereof.
5. The method of claim 4 , wherein the viruses are selected from the group consisting of influenza viruses, measles and respiratory syncytial viruses, dengue viruses, human immunodeficiency viruses, human hepatitis viruses, herpes viruses, papilloma viruses, and wherein the protozoa is Plasmodium falciparum , and the bacteria is tuberculosis-causing Mycobacteria.
6. The method of claim 4 , wherein the tumor associated antigen is selected from hematological malignancies and solid tumors.
7. The method of claim 6 , wherein said solid tumors are selected from the group consisting of colon carcinoma, breast carcinoma, prostate carcinoma, renal cell carcinoma (RCC), lung carcinoma, sarcoma and melanoma.
8. The method of claim 1 , wherein the selecting in step d) comprises measuring the cytokine secretion of the T cells or other measures of T cell activation.
9. The method of claim 1 , further comprising expanding the antigen-specific T cells selected in step d) ex vivo.
10. The method of claim 1 , further comprising cloning the T cell receptor (TCR) of the antigen-specific T cells selected in step d).
11. The method of claim 10 , further comprising expressing the TCR as a transgene in a PBMC.
12. The method of claim 1 , wherein the nucleic acid encoding the patient-derived MHC molecule and the antigen is provided as a bicistronic RNA encoding both the patient-derived MHC molecule and the antigen.
13. The method claim 1 , wherein the MHC molecule is an MHC class I molecule or is an MHC class II molecule.
14. The method of claim 13 , wherein the MHC class I molecule is selected from the group consisting of HLA-A, HLA-B, HLA-C and HLA-E.
15. The method of claim 13 , wherein the MHC class II molecule is selected from the group of HLA-DP, HLA-DQ and HLA-DR.
16. The method of claim 1 , wherein the APCs derived from the healthy donor are dendritic cells.
17. A method of generating antigen T cells specific for a particular HLA-A, HLA-B, HLA-C or HLA-E class I MHC molecule bound to a solid tumor cell associated antigen, comprising:
a) providing a nucleic acid encoding an HLA-A, HLA-B, HLA-C or HLA-E class I MHC molecule from a patient having a solid tumor, and further providing a solid tumor cell associated antigen or nucleic acid encoding a solid tumor cell associated antigen;
b) co-transfecting or introducing both components as defined in (a) into dendritic cells derived from a healthy donor who is allogeneic for the HLA-A, HLA-B, HLA-C or HLA-E class I MHC molecule;
c) priming peripheral blood lymphocytes (PBLs) derived from the healthy donor with the dendritic cells; and
d) using a tetramer comprising the particular HLA-A, HLA-B, HLA-C or HLA-E class I MHC molecule bound to a peptide of the solid tumor cell associated antigen to select for those T cells which are specific for the particular HLA-A, HLA-B, HLA-C or HLA-E class I MHC molecule bound to the solid tumor cell associated antigen.