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. 2022 May 16;10(5):e003819. doi: 10.1136/jitc-2021-003819

Figure 1.

Figure 1

ABZ enhances CTL activity in association with decrease of tumor PD-L1 expression. (A) Representative western blotting of PD-L1 protein level in A375, SK-MEL-28 and A549 cells treated with increasing concentrations of ABZ (0.625–1.25 µM) for 24 hours under IFN-γ exposure. (B) Representative profiles and quantitative analysis of membrane PD-L1 expression by flow-cytometric analysis after increasing concentrations of ABZ (0.625–1.25 µM) treated A375, SK-MEL-28 and A549 cells for 24 hours under IFN-γ exposure. (C) A375 and SK-MEL-28 cells co-cultured with activated T cells for 24 hours with or without ABZ (0.625–1.25 µM) were subjected to crystal violet staining. The tumor cell to T cell ratio, 1:3. The quantitative analysis of A375 and SK-MEL-28 cell survive rate from three independent experiments and showed as means±SD, *p<0.05, **p<0.01, ***p<0.001. (D–G). C57BL/6 mice were implanted with 1×106 B16F10 and received 50 mg/kg of ABZ treatment. (D) A schematic view of the treatment plan. (E) Tumor volume was measured on the indicated different time points. (F, G) Photographs of representative tumors and tumor weight were measured after ABZ treatment on day 12 in the B16F10 tumor burden mouse model. Data represent mean±SD, ***p<0.001. (H-K) C57BL/6 mice were implanted with 1×106 LLC and received 50 mg/kg of ABZ treatment. (H) A schematic view of the treatment plan. (I) Tumor volume was measured on the indicated different time points. (J, K) Photographs of representative tumors and tumor weight were measured after ABZ treatment on day 15 in the LLC tumor burden mouse model. Data represent mean±SD, **p<0.01. (L, M) Representative profiles and quantification of flow cytometry-based detection of the GZMB+ CD8+ in B16F10 and LLC tumor mass from the different treatment groups (n=5 mice per group). Data represent mean±SD, ***p<0.001. (N, O) Representative profiles and quantification of flow cytometry-based detection of the PD-L1 in B16F10 and LLC tumor mass from the different treatment groups (n=5 mice per group). Data represent mean±SD, ***p<0.001. ABZ, albendazole; CTL, cytotoxic T lymphocyte; IFN-γ, interferon gamma; LLC, Lewis lung cancer.