Supplementary Figure 12: Quantitation of cytokine and T cell exhaustion markers of Cpf1 CD22BBz KIKO CAR-T cells compared to Cas9-mediated CD22BBz;PDCD1–/– CAR-T cells.
From: One-step generation of modular CAR-T cells with AAV–Cpf1
a, FACS quantitative analysis of antigen-specific IFN-γ and TNF-α production capability in Cpf1 CD22BBz KIKO versus Cas9 RNP CD22BBz CAR-T cells (n = 3 infection replicates each). Two-way ANOVA with Tukey’s multiple-comparisons test was used to assess significance (multiple-testing corrected). IFN-γ group: vector versus Cas9, ***P < 0.001; vector versus Cpf1, ***P < 0.001; Cas9 versus Cpf1, P = 0.4835. TNF-α group: vector versus Cas9, ***P < 0.001; vector versus Cpf1, ***P < 0.001; Cas9 versus Cpf1, P = 0.1318. b, FACS quantitative analysis of Cpf1 CD22BBz KIKO versus Cas9 RNP CD22BBz CAR-T cells in T cell exhaustion, showing that the expression of exhaustion markers (PD-1, TIGIT, and LAG3) in Cpf1 CD22BBz KIKO CAR-T cells was lower than that in Cas9 RNP CD22BBz CAR-T cells (n = 3 infection replicates each). Two-way ANOVA with Tukey’s multiple-comparisons test was used to assess significance (multiple-testing corrected). PD-1 group: vector versus Cas9, ***P < 0.001; vector versus Cpf1, P = 0.9087; Cas9 versus Cpf1, ***P < 0.001. TIGIT group: vector versus Cas9, ***P < 0.001; vector versus Cpf1, ***P < 0.001; Cas9 versus Cpf1, ***P < 0.001. LAG3 group: vector versus Cas9, ***P < 0.001; vector versus Cpf1, ***P < 0.001; Cas9 versus Cpf1, ***P < 0.001. All data are shown as mean ± s.e.m., plus individual data points on the graph.
