Figure 6. The enhanced cytokine production by the Cas9-mediated hPD-1 KO in primary T cells.

sgRNA hPD-1:Cas9 modified primary T cells and control T cells from healthy donors or patients were cultured in IL-2 after electroporation for 7 d and stimulated by peptides pulsed autologous DCs for 20 h. The IFN- γ secreting cells was evaluated by Elispot. (a,b) T cells from two EBV serum positive healthy donors were stimulated by LMP2a peptides pulsed autologous DCs. (c) T cells from a melanoma patient was stimulated by melanoma associated peptides pulsed autologous DCs. (d) T cells from a gastric cancer patients were stimulated by gastric cancer associated peptides pulsed autologous DCs. All values shown are mean ± SD of triplicate or duplicate measurements and have been repeated 3 times with similar results. *p < 0.05; **p < 0.005.