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. 2023 Mar 10;62(4):53. doi: 10.3892/ijo.2023.5501

Figure 3.

Figure 3

Double-target CAR-T cells show cytolytic potency to target HCC cells. (A) Cytotoxicity of GPC3/PD-1-CAR-T cells to the four indicated HCC cell lines at various E:T ratios evaluated by an LDH cytotoxicity assay. (B) Comparison of the toxicity of GPC3/PD-1-CAR-T cells to HuH7 cells with different GPC3 levels at E:T ratios of 4:1, 8:1, 16:1 and 32:1. Un-transduced T cells served as control. (C) Comparison of the toxicity of various CAR-T cells against HuH7 at different E:T ratios. Un-transduced T cells served as control. (D) Levels of GrB and perforin measured using ELISA after co-culturing different CAR-T cells with HuH7 at an E:T ratio of 1:1 for 24 h. (E) Comparison between the cytotoxicity of GPC3/PD-1-CAR-T cells and that of HuH7, PD-1+ T and T cells at various E:T ratios, as assessed by an LDH assay. (F) The death rate of HuH7 cells under a series of concentrations of GrB and perforin for 12 h. (G) The death rates of HuH7, PD-1+ T and T cells after co-culturing in the indicated culture medium for 12 h. Data are presented as the mean ± SD. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; CAR, chimeric antigen receptor; HCC, hepatocellular carcinoma; GPC3, glypican-3; PD-1, programmed death 1; E:T, effector-to-target; LDH, lactate dehydrogenase; GrB, granzyme B.