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Strategies that enhance the function of T cells are critical for immunotherapy.
Here we described for the first time a non-viral mediated approach to reprogram primary human T cells by disruption of PD-1.
We showed that the gene knockout of PD-1 by electroporation of plasmids encoding sgRNA and Cas9 was technically feasible. The disruption of PD-1 resulted in significant reduction of PD-1 expression but didn't affect the viability of primary human T cells. Cellular immune response of the gene modified T cells was characterized by up-regulated IFN-γ production and enhanced cytotoxicity.
These results suggest that we have established an approach for efficient checkpoint inhibitor disruption, providing a new strategy for targeting checkpoint inhibitors to improve the efficacy of T cell based adoptive therapies.
The authors thank the entire Huang Lab and Liu Lab for their support and advice. This work was funded by grants from the National Natural Science Foundation of China (Grant No. 81172281, 81000980, 81220108023, 81172094).