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. 2021 Mar 22;12:661357. doi: 10.3389/fimmu.2021.661357

Figure 2.

Figure 2

IDO1 accumulation in ESCC may restrict the effectiveness of CAR-T cell therapy. (A) Western blot analysis of IDO1 and β-actin levels in ESCC cells treated with or without IFN-γ. (B) Western blot analysis of IDO1 and β-actin levels in EC1 cells stably expressing shControl or shIDO1. (C) Apoptosis was determined by Annexin-V/PI staining of shControl or shIDO1 cells after 6 h of CAR-T cell culture. (D) Schematic of the experiments using SCID/Beige mice, comparing the antitumor effects of MSLN-CAR-T cells in the groups treated with EC1-shControl or EC1-shIDO1 cells. (E) Tumor volume was evaluated for 15 d. Expression of (F) PD-1,TIM-3, (G) CD28, and CD69 in tumor-infiltrated CAR-T cells. (H) Proportions of IFN-γ, IL-2, and perforin in tumor-infiltrated CAR-T cells. *P < 0.05, **P < 0.01, ***P < 0.001, ns, Not statistically significant (repeated-measures one-way ANOVA or Student’s t-test).