Extended Data Fig. 7: IL-1α expression is neither regulated by active caspase-1 in human Th17 cells nor coregulated with IL-1β, in contrast to the situation in monocytes.
From: Human TH17 cells engage gasdermin E pores to release IL-1α on NLRP3 inflammasome activation
a, Th17 cells were isolated ex vivo by FACS-sorting and stimulated for 5 days with anti-CD3 and anti-CD28 mAbs (48 h plate-bound) in the presence or absence of the indicated cytokines before flow cytometric analysis of intracellular IL-1α and active caspase-1 after PMA and ionomycin restimulation. The results of one representative experiment are shown. b, Cumulative data for active caspase-1 as shown in (a). One-way ANOVA. n = 3 biologically independent samples examined over 3 independent experiments. Data are presented as mean ± SEM. c, Intracellular staining and flow cytometry of monocytes following 24 h stimulation with LPS and 30 min with nigericin (Nig.). The results shown are from one representative experiment. d, e, cumulative data of (d), Two-sided paired t-test. Each circle indicates an individual healthy blood donor (n = 3-4 biologically independent samples examined over 3 independent experiment). Data are presented as mean ± SEM. f, ELISA of Th17 cells, which were stimulated as in (a) and of monocytes were stimulated with LPS (24 h) and ATP (30 min). n = 3 biologically independent samples examined over 2 independent experiments. Data are presented as mean ± SEM. g, Flow cytometric analysis of cells as in (g).h, Intracellular cytokine staining and flow cytometric analysis of T-cell clones generated from the Th17-cell subset. Each circle indicates an individual T cell clone (n = 64). Data are presented as mean ± SEM. i, scRNA-seq and UMAP showing IL-1β expression in human Th17 cells stimulated for 5 days with anti-CD3 and anti-CD28 mAbs.
