Fig. 6

TGFβ-producing T_RM precursors are found mainly within circulating CD8⁺ PD1⁺CD28⁺ T cells. A Left, representative flow cytometry dot-plots showing CD103 versus CD69 staining, in ex vivo CD8⁺ T cells isolated from PBMC, adjacent non-tumor tissue (NT) and tumor site (T), in one NSCLC patient. Right, quantification of ex vivo CD8⁺ T_RM (CD103⁺CD69⁺) cells in 25 NSCLC patients (PBMC, n = 19; NT, n = 14; T, n = 25) (Mann–Whitney test with Bonferroni correction for multiple comparisons). B Representative flow cytometry dot-plots showing the proportion of T_RM cells in gated CD8⁺ PD1⁺CD28⁻ or PD1⁺CD28⁺ T-cell subsets, in tumor sites. C Quantification of T_RM cells in total CD8⁺ or in gated PD1⁺CD28^- or PD1⁺CD28⁺ cells, in NT (n = 14) and tumor sites (n = 25) (Wilcoxon rank test with Bonferroni correction). D Percentage of CD103 expression in the different T-cell subsets from ex vivo PBMC (n = 15), NT (n = 15), and tumor site (n = 26) of NSCLC patients (Wilcoxon rank test, with Bonferroni correction for multiple comparisons). E Representative flow cytometry dot-plots showing CD103 versus CD69 staining, in circulating PD1/CD28 T cells from one HD. F Percentage of CD103 expression in PD1/CD28 T-cell subsets from HDs (n = 9) (Wilcoxon rank test). G Representative flow cytometry dot-plots showing PD1 vs CD28 staining, before and after FACS-sorting, in one HD. H Expression of CD103 in sorted PD1⁺CD28⁻ or PD1⁺CD28⁺ T cells, following 6-days of in vitro expansion by stimulation with anti-CD3 mAb, in the presence or absence of TGFβ (Wilcoxon rank test). I Intracellular TGFβ production, evaluated by flow cytometry, in the different T-cell subsets as indicated, with or without anti-CD3 mAb activation (5h), in 6 HDs (Wilcoxon rank test). * P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. NS, not significant. Plots show median values with interquartile range