US 12,606,831 B2 · Method for producing t cells modified by chimeric antigen receptor

Abstract

A method for producing γδ T cells modified by a chimeric antigen receptor, comprising: transfecting K562 cells with shFPPS targeted to FPP synthase by means of a lentiviral vector, such that the expression of FPPS in the K562 cells is lowered and a K562-shFPPS cell line with reduced FPPS expression is constructed; adding the K562-shFPPS cell line into a γδ T cell culture system for co-culturing with the γδ T cells, wherein it is found that the K562-shFPPS cell line can facilitate in vitro differentiation and proliferation of the γδ T cells; and adding a CAR-expressing lentiviral vector to the γδ T cell culture system comprising the cell line for co-culturing, wherein it is found that the K562-shFPPS cell line can further effectively improve the transfection rate of CAR genes. The provided solution effectively overcomes the technical challenge of the large-scale production of CAR-γδ T cells, and has a good application prospect.

Claims9

Dependent claims

A method for expanding γδ T cells, comprising:

introducing shRNA that inhibits the expression of FPPS (Farnesyl pyrophosphate Synthase)-encoding gene into tumor cells to obtain tumor cells with reduced FPPS expression and activity;
adding the tumor cells with reduced FPPS expression and activity to a γδ T cell culture system,
wherein the shRNA that inhibits the expression of FPPS-encoding gene is single-stranded RNA comprising in 5′ to 3′ order:
a stem I, a loop and a stem II;
wherein the stem I consists of the sequence of positions 1-21 of SEQ ID NO:
2;
the loop consists of the sequence of positions 22-27 of SEQ ID NO:
2;
and the stem II consists of the sequence of positions 28-48 of SEQ ID NO:
2.

7 more elements

Dependencies are inferred from the claim text.

Method for producing t cells modified by chimeric antigen receptor

Life of this patent

Method for producing t cells modified by chimeric antigen receptor

Life of this patent

Abstract

Claims9