Fig. 3.

Differences in cytotoxicity activation molecules and cell surface marker expression between CD8⁺PD-1⁺TCF1⁺ progenitor exhausted T cells (TCF1⁺Texprog) and CD8⁺PD-1⁺TCF1⁻ terminally exhausted T cells (TCF1⁻Tex^term) in head and neck squamous cell carcinoma (HNSCC) tumor tissues were detected using flow cytometry. a Representative contour plots of selected cytokine markers (TNFα, GZMB, and IFNγ) gated on TCF1⁺Tex^prog or TCF1⁻Tex^term. b TCF1⁻Tex^term showed higher GZMB production (n = 20). c There was no significant difference in IFNγ expression between these two subsets of exhausted T cells (n = 20). d TCF1⁺Tex^prog showed a higher TNFα production (n = 20). e TCF1⁻Tex^term displayed a higher proportion of cells with the polyfunctional phenotype of TNFα⁺GZMB⁺IFNγ⁺ (n = 20). f Representative contour plots of CD103 and comparison of CD103 expression among TCF1⁺Tex^prog and TCF1⁻Tex^term (n = 24). g Mean fluorescence intensity (MFI) of TIM-3, CTLA-4, TIGIT, and LAG-3 gated on TCF1⁺Tex^prog or TCF1⁻Tex^term. h Quantitative analysis of TIM-3 (n = 24), CTLA-4 (n = 10), TIGIT (n = 10), and LAG-3 (n = 10) MFI between TCF1⁺Tex^prog and TCF1⁻Tex^term. Paired t tests were performed for (b–f and h). *P < 0.05, **P < 0.01, ****P < 0.000 1, ^nsP > 0.05