Fig. 5: Notch/STAT3-mediated acquisition of IL-10 expression is defective in Crohn’s disease patients.

a Effector memory CD4⁺ T cells were isolated from blood of healthy donors (HD), Crohn’s disease (CD) or ulcerative colitis (UC) patients. Afterwards, cells were cultured and co-stimulated w/ or w/o DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) for 120 h and restimulated with PMA/Ionomycin for optimal cytokine detection. a Quantification of IL-10⁺ cells (left) and representative flow cytometric profiles of CD4 vs. IL-10 expression after DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation (right). b Quantification of IL-10⁺ cells among Th1 (IFN-γ⁺), Th17 (IL-17A⁺) and Th1/17 (IFN-γ⁺IL-17A⁺) cells from HD, CD and UC patients after DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) costimulation (left). Representative flow cytometric profiles of CD4 vs. IL-10 expression after DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation (right). c Quantification of IFN-γ⁺, IL-17A⁺ and IFN-γ⁺IL-17A⁺ cells among effector memory CD4⁺ T cells from HD, CD and UC patients co-stimulated w/ or w/o DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) (left). Representative flow cytometric profiles of IFN-γ vs. IL-17A expression after DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21) co-stimulation (right). d Ratio of IFN-γ⁺, IL-17A⁺ and IFN-γ⁺IL-17A⁺ cells to IL-10⁺ cells (frequency) among effector memory CD4⁺ T cells from HD, CD and UC patients co-stimulated w/ or w/o DLL4/Cyt (Cyt = IFN-α/IL-6/IL-21). Each dot represents one donor (n_HD=19, n_CD = 11, n_UC = 9). Two-tailed Mann-Whitney and unpaired t tests were performed (* P < 0.05; ** P < 0.01; *** P < 0.001) to assess significance. Data are cumulative from nine independent experiments.