Extended Data Fig. 9: Tumor hypoxia enforces CD39 expression on tTex cells.
From: Hypoxia drives CD39-dependent suppressor function in exhausted T cells to limit antitumor immunity
(a) Histogram overlay displaying hypoxia exposure in CD8+ dLN and TIL from B16-F10 tumors. (b) CD39 staining in exhausted T cells from B16-F10 or MC38, and (c) Pimonidazole staining of bulk TIL. (d) Continuous Activation under Hypoxia (CS + H) assay. In brief, naïve T cells are activated for 24 hours, then split into treatment groups of removal (acute stimulation) or continued presence (continuous stimulation) of anti-CD3/anti-CD28-bound microbeads and cultured under atmospheric oxygen tensions (~20% O2) or tumor hypoxic conditions (1.5% O2) for 5 days. (e) Inhibitory receptor staining in murine day 6 CS + H cells. (f–h) Validation of humanized CS + H assay with healthy donor PBMC-derived CD8+ T cells via staining of (F) inhibitory receptors and (g) enzymes of adenosine metabolism. (h) 24-hour PMA/Ionomycin re-stimulation of human CS + H cells. Statistics are Mann-Whitney (B,C), one-way ANOVA (H) with ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 and ∗∗∗∗p < 0.0001.
