Methods for engineering allogeneic and highly active t cell for immunotherapy
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. 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) with a 4-1BB intracellular activation domain and have at least one PD1 allele inactivated;
wherein the CAR comprises the amino acid sequence of SEQ ID NO:73, and
wherein the inactivation of the at least one PD1 allele enhances the anti-tumor activity of the population of isolated primary human T cells expressing the CAR.
2. The population of isolated primary human T cells of claim 1 , further having the CTLA4 alleles in the cells inactivated.
3. The population of isolated primary human T cells of claim 1 , having the TCRα alleles and the CD52 alleles in the cells inactivated.
4. The population of isolated primary human T cells of claim 3 , further expressing a fragment of preTalpha sufficient to support CD3 surface expression.
5. The population of isolated primary human T cells of claim 1 , having the CD52 alleles in the cells inactivated.
6. The population of isolated primary human T cells of claim 1 , having the TCRα alleles in the cells inactivated.