Fig. 4

Functional benefit provided by CD11a^high, ICOS and CD137 depends on the PD1/CD28 phenotype and localization. A Expression levels of co-stimulatory receptors and lytic molecules in T cells from PBMC, adjacent non-tumor tissue (NT), and tumor site (T), in a representative NSCLC patient, illustrated by t-SNE plots. B-E–H Expression of CD11a^high (B, n = 23), ICOS (E, n = 21) and CD137 (H, n = 38), as evaluated by multiparametric flow cytometry, within ex vivo unstimulated T cells gated on CD8⁺ PD1⁺CD28⁻ and PD1⁺CD28⁺ subsets, from the periphery to the tumor site of NSCLC patients (Mann–Whitney test, with Bonferroni correction for multiple comparisons). C-F-I Expression of CD11a^high (C), ICOS (F), and CD137 (I), in the comparison between CD8⁺ PD1⁺CD28⁻ and PD1⁺CD28⁺ T cells within the same district (Wilcoxon rank test). D-G-L Polyfunctional ability in CD8⁺ PD1⁺CD28⁻ or PD1⁺CD28⁺ T-cell subsets following activation with anti-CD3 mAb (5-6h), between gated CD11a^high vs CD11a^low/− (D, n = 10), ICOS⁺ vs ICOS⁻ (G, n = 6) and CD137⁺ vs CD137⁻ (L, n = 10) cells, in the different districts of matched patients (Wilcoxon rank test). * P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. NS, not significant. Plots show median values with interquartile range