Fig. 8: The Mouse Orthotopic Transplant Model showed that the combination of PD-1 and TIM-3 monoclonal antibodies enhances the efficacy of immunotherapy in HNSCC.

C3H mice were injected with 2 × 10⁵ SCC-7 cells overexpressing circE7 or vector control in the right cheek to form xenograft tumors. After 3 days, 20 μg/each of anti-PD-1 or anti-PD-1/TIM-3 were injected for immunotherapy. A Representative tumor images. (n = 7 for each group). B Tumor volume measured 3 days, 7 days and 10 days after SCC-7 cell injection (n = 7 for each group). C Tumor weight measured 10 days after SCC-7 cell injection (n = 7 for each group). D Prepare single-cell suspension from mouse tumor tissue, RT-qPCR detection of IFNG, GZMB, and TNFA mRNA expression (n = 3 per group). E ELISA was used to detect the content of IFN-γ, GZMB, and TNF-α (n = 3 per group). Flow cytometry was performed to detect the proportion of CD8⁺ T cells (F) and the percentage of IFN-γ⁺ TNF-α⁺ cells (G) within the total CD8⁺ T cell population (n = 3 for each group). H Immunohistochemistry was performed to detect the expression of Cd8a in SCC-7 xenograft tumors, and the results were statistically analyzed (n = 5 for each group). I Immunohistochemistry was performed to detect the expression of galectin-9 in paraffin-embedded SCC-7 xenograft tumors, and the results were statistically analyzed (n = 5 for each group). Correlations were calculated using unpaired two-tailed Student’s t test for (B, C, D, E, F, G, H, I), *p < 0.05, **p < 0.01, and ***p < 0.001. Data are presented as mean ± SD. Source data are provided as a Source Data File.