Fig. 5: ILC3s promote OSMR expression in IECs in H. hepaticus-induced colitis.

a,b, H. hepaticus + anti-IL-10R colitis was induced in wild-type mice for 7 days. Percentage (a) and absolute cell numbers (b), respectively, of IL-22- or Ki67-expressing CD45⁺Lin^– (that is, live CD45⁺CD19^–CD11c^–Ly6G/C^–F4/80^–FcεRIα^–) LPMCs in naive mice and colitic mice were assessed. Flow cytometry-assessed cytokine expression after stimulation with phorbol 12-myristate 13-acetate (PMA)/ionomycin, IL-23 and IL-1β is shown. Data are representative of two experiments; n = 12. c, Representative gating strategy for identifying αβ and γδ T cells and different ILC subsets in colitic mice. ILC subsets were identified by flow cytometry of mouse LPMCs. The ILC population was characterized as CD45⁺Lin^– (that is, CD19^–CD11c^–Ly6G/C^–F4/80^–FcERIα^–TCRα^–TCRγ^–) and live cells. Subsets were further defined based on transcription factor expression: ILC3 (RORγt⁺Lin^–TCRα^–TCRγ^–), ILC2 (GATA3⁺RORγt^–T-bet^–Eomes^–Lin^–TCRα^–TCRγ^–), ILC1 (T-bet⁺Eomes^–RORγt^–Lin^–TCRα^–TCRγ^–) and natural killer cells (T-bet⁺Eomes⁺RORγt^–Lin^–TCRα^–TCRγ^–). d, Pie chart showing the indicated cell populations within total IL-22⁺CD45⁺CD19^–CD11c^–Ly6G/C^–F4/80^–FcεRIα^– cells in steady-state and inflamed mice on day 7 of colitis. e,f, IL-22 production by different ILC populations identified as described in c. Data are representative of two experiments; n = 12. P values are derived from Mann–Whitney U-tests. g, H. hepaticus + anti-IL-10R colitis was induced in wild-type mice. Mice were treated with anti-IL-12p40, anti-IL-23p19 or isotype control on day 0 and analyzed on day 7. Osmr (left) and Reg3g (right) expression was quantified in epithelial cells by qPCR. Data are representative of one to two independent experiments; n = 6–29 per time point. Adjusted P values were obtained using a Kruskal–Wallis test followed by a Dunn’s multiple comparisons test. h–j, H. hepaticus + anti-IL-10R colitis was induced in Rag2^–/–Rorc^–/– and Rag2^–/– mice for 7 days. h,i, qPCR gene expression analysis of Osmr (h) and Il22 and Ifng (i) in colonic IECs from inflamed Rag2^–/–Rorc^–/– and Rag2^–/– mice normalized to steady-state mice. j, Histopathological scoring of colitis in Rag2^–/–Rorc^–/– and Rag2^–/– mice on day 7 of colitis. Data are from one to two independent experiments; n ≥ 6. P values (two-tailed) were calculated using a Mann–Whitney test in a, b, f and h–j.