Fig. 6: Combination treatment with exogenous TIM-3 and PD-1 monoclonal antibodies enhances CD8⁺ T cell cytotoxic function.

CAL27 and HN30 cells were overexpressed with circE7, or SCC090 and SCC154 cells were knocked down with circE7, and then co-cultured with primary human T cells with the addition of 5 μg/mL anti-PD-1 or anti-PD-1/TIM-3 for 6 h. A. RT-qPCR was used to measure the expression levels of IFNG, GZMB, and TNFA mRNA in CD8⁺ T cells. B. Flow cytometry was used to detect the proportion of IFN-γ⁺ GZMB⁺ and IFN-γ⁺ TNF-α⁺ T cells among all CD8⁺ T cells. Left panel shows representative images; right panel shows the statistical results, as shown in Supplementary Fig. S9B. C, D Annexin V/PI staining was used to assess the apoptosis levels of CD8⁺ T cells (C) and tumor cells (D), as shown in Supplementary Fig. S9C, D. Correlations were calculated using unpaired two-tailed Student’s t test for (A, B, C, D), *p <  0.05, **p < 0.01, and ***p < 0.001. These experiments were derived from three independent repetitions. Data are presented as mean ± SD. Source data are provided as a Source Data File.