Fig. 2: Notch/STAT3-induced IL-10 expression is transient and independent of CD4+ TH cell specification. | Mucosal Immunology

Fig. 2: Notch/STAT3-induced IL-10 expression is transient and independent of CD4+ TH cell specification.

From: A Notch/STAT3-driven Blimp-1/c-Maf-dependent molecular switch induces IL-10 expression in human CD4+ T cells and is defective in Crohn´s disease patients

Fig. 2

a Memory CD4+ T cells were cultivated w/ or w/o DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation for 120 h (1st wk). Afterwards cells were rested and re-stimulated for additional 120 h (2nd wk) without co-stimulation. After each round of stimulation, cells were restimulated with PMA/Ionomycin and subsequently stained for IL-10 and IFN-γ. Quantification of two independent experiments is shown. b FACS-isolated Th1, Th2 and Th17 cells were cultivated in vitro w/ or w/o DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation for 120 h and afterwards restimulated with PMA/Ionomycin for optimal cytokine detection. Representative flow cytometric profiles of IL-10 vs. IFN-γ, IL-4 and IL-17A expression are shown. c Quantification of IL-10+ cells among Th1, Th2, Th17 and Th1/17 cells cultivated in vitro w/ or w/o DLL4, Cyt or DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation. d Quantification of IL-10+ cells among IFN-γ+/− (Th1, Th1/17), IL-4+/− (Th2), IL-17A+/− (Th17, Th1/17) cells cultivated in vitro w/ or w/o DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation. e Quantification of IFN-γ+, IL-4+ and IL-17A+ cells among FACS-isolated Th1, Th2, Th17 and Th1/17 cells, respectively, after in vitro cultivation and w/ or w/o DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation. Each dot represents one healthy donor (n = 7–32). Two-tailed Wilcoxon, Mann–Whitney and unpaired t tests were performed (*P < 0.05; **P < 0.01; ***P < 0.001) to assess significance. Data are cumulative from up to eight independent experiments.

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