Fig 1: Hypoxia induces dysfunction and terminal exhaustion of human T cells.a Schematic graph demonstrating the coculture model. b Representative flow cytograms (upper panel) gated from human pan-T cell culture and quantification (lower panel, n = 3) of differentiated CD8+ T cell subtypes: Tn (naïve T cells), Tcm (central memory T cells), Tem (effector memory T cells), Teff (effector T cells). c Schematic graph demonstrating the normoxia (20% O2) and hypoxia (1% O2) culture condition of T cells coculturing with human TNBC cell line. d Heatmap of the differentially expressed genes (DEGs) in hypoxic cultured human T cells compared to normoxia group. DEGs were identified in edgeR (|logFC| > 1, adjusted P < 0.01). P values were adjusted using Benjamini–Hochberg method in edgeR. DEGs identified in the indicated GO gene clusters are marked in the heatmap. e GSEA analysis of human T cells in hypoxic versus normoxic conditions. Analysis was based on ranked logFC from edgeR. FDR and adjusted p value are shown in the graph. P values were adjusted using Benjamini–Hochberg method in GSEA analysis. f Flow cytometry quantifications of immune effector molecules and exhaustion markers in CD8+ T cells gated from human pan-T cells cultured under the indicated conditions (n = 4). g Representative flow cytograms of PD-1 and TIM-3 expression in CD8+ T cells gated from human pan-T cells culture. h Flow cytometric quantification of terminally exhausted T cells (PD-1+ TIM-3+) in CD8+ T cells gated from human pan-T cells culture (n = 3). i Flow cytometric quantification of proliferating cells (Ki76+) in CD8+ and CD4+ T cells gated from human T cells cocultured with TNBC (n = 3). All flow cytometry data (b, f, h, and i) are presented as the mean ± SD of samples from three to four donors. For all flow cytometry data, P values were determined by one-way ANOVA (f, h) or two-way ANOVA (b) with Turkey’s test, or paired two-tailed t-test (i). Raw RNA-seq data is available in the GEO database with accession number GSE179885. For the remaining data, source data are provided in Source Data file.