METHODS FOR ENGINEERING ALLOGENEIC AND HIGHLY ACTIVE T CELL FOR IMMUNOTHERAPHY
The present invention relates to methods for developing engineered T-cells for immunotherapy that are non-alloreactive. The present invention relates to methods for modifying T-cells by inactivating both genes encoding T-cell receptor and an immune checkpoint gene to unleash the potential of the immune response. This method involves the use of specific rare cutting endonucleases, in particular TALE-nucleases (TAL effector endonuclease) and polynucleotides encoding such polypeptides, to precisely target a selection of key genes in T-cells, which are available from donors or from culture of primary cells. The invention opens the way to standard and affordable adoptive immunotherapy strategies for treating cancer and viral infections.
1 - 28 . (canceled)
29 . A method of treatment comprising administering a population of isolated primary human T cells comprising at least 10 5 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
30 . The method of claim 29 , wherein the population of isolated primary human T cells has the CTLA4 alleles in the cells inactivated.
31 . The method of claim 29 , wherein the population of isolated primary human T cells has the TCRα alleles and the CD52 alleles in the cells inactivated.
32 . The method of claim 29 , wherein the population of isolated primary human T cells express a fragment of preTalpha sufficient to support CD3 surface expression.
33 . The method of claim 29 , wherein the population of isolated primary human T cells has the CD52 alleles in the cells inactivated.
34 . The method of claim 29 , wherein the population of isolated primary human T cells has the TCRα alleles in the cells inactivated.
35 . The method of claim 29 , wherein the population of isolated primary human T cells has both PD1 alleles in the cells inactivated.
36 . The method of claim 29 , wherein the population of isolated primary human T cells comprises at least 10 6 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
37 . The method of claim 30 , wherein the population of isolated primary human T cells comprises at least 10 6 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
38 . The method of claim 31 , wherein the population of isolated primary human T cells comprises at least 10 6 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
39 . The method of claim 33 , wherein the population of isolated primary human T cells comprises at least 10 6 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
40 . The method of claim 34 , wherein the population of isolated primary human T cells comprises at least 10 6 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
41 . The method of claim 35 , wherein the population of isolated primary human T cells comprises at least 10 6 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
42 . The method of claim 29 , wherein the population of isolated primary human T cells comprises at least 10 7 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
43 . The method of claim 30 , wherein the population of isolated primary human T cells comprises at least 10 7 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
44 . The method of claim 31 , wherein the population of isolated primary human T cells comprises at least 10 7 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
45 . The method of claim 33 , wherein the population of isolated primary human T cells comprises at least 10 7 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
46 . The method of claim 34 , wherein the population of isolated primary human T cells comprises at least 10 7 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.
47 . The method of claim 35 , wherein the population of isolated primary human T cells comprises at least 10 7 isolated primary human T cells that express a chimeric antigen receptor (CAR) comprising the amino acid sequence of SEQ ID NO:73 and have at least one PD1 allele inactivated.