Fig. 1: Repetitive antigen-specific stimulation ex vivo of T cells results in exhaustion.
From: Deletion of SNX9 alleviates CD8 T cell exhaustion for effective cellular cancer immunotherapy
a Scheme representing the principle of the exhaustion model. b Representative co-expression of PD-1 and TIM-3 measured by antibody staining and flow cytometry. c PD-1 expression measured by flow cytometry after antibody staining on day 12 after the first stimulation. n = 10 donors from n = 5 experiments except for Teff n = 4 of n = 2 experiments. d Degranulation measured by CD107a surface exposure during re-stimulation of all cells after 13 days of culture. n = 12 donors of n = 6 experiments except for Teff n = 6 of n = 3 experiments. e–f Representative flow cytometry plot and quantification of IFNγ and TNFα production measured by intracellular cytokine staining. n = 14 donors from n = 7 experiments except for Teff n = 8 of n = 4 experiments. g Specific lysis of T2-Luc+ tumor cells pulsed with peptide after 5 hours of co-incubation with T cells measured by luminescence intensity. n = 6 donors for Trest, n = 13 for Tex and n = 8 for Teff. h IFNγ secretion measured by ELISA after 4 days co-culture of Teff or Tex with MDA-MB-231 cells loaded with 100 nM NY-ESO-1-9V peptide in presence of either 10 μg/ml human IgG4 isotype control or anti-PD-1 antibody (Nivolumab). Statistics is a paired 2-way-ANOVA with Holm-Sidak correction with n = 5 donors. i Heatmap of all differentially regulated genes between the four conditions showing row-scaled log-cpms and clustered using k-means (numbers indicated on the left). Selected genes associated with exhaustion or functionality are labeled on the right. n = 4 donors. j Gene set enrichment analysis comparing mRNA expression of the different conditions for the Zheng. et al PanCancer T cell states. Color represents the mean log fold change and the size the -log10 false discovery rate (FDR). c-d; f-g 1-way ANOVA statistics with Holm-Sidak correction, Mean and SD are shown. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Source data and exact p-values are provided as a Source Data file.
