αIL-1β treatment reduces the accumulation of IL-17A–producing ILCs in the colon. (A) IL-1R1 expression by Sca1⁺Thy1.2^Hi ILCs and rest of large intestine lamina propria cells. 129SvEv Rag2^−/− mice were infected with H. hepaticus and sacrificed after >8 wk of infection. LPLs were isolated from the large intestine, Sca1⁺Thy1.2^Hi ILCs were FACS sorted, and Il1r1 expression was evaluated by qRT-PCR (mean ± SEM, n = 2 from 2 independent experiments; 8–10 mice were pooled in each experiment). (B–E) 129SvEv Rag2^−/− mice were infected with H. hepaticus and treated weekly with 1 mg of αIL-1β antibody or isotype control (i.p.). After 8 wk, mice were sacrificed and cLPLs were isolated. (B) Total numbers of Sca1⁺ Thy1.2^Hi ILCs in the colon lamina propria as evaluated by FACS analysis. (C) Total numbers of IL-17A– or IFN-γ–producing ILCs from the colon of indicated mice groups. (D) Cytokine production by cLPLs after overnight culture in complete medium alone (n/a) or in the presence of 10 ng/ml IL-23. Data are represented as mean ± SEM from 2 pooled independent experiments (n = 6–11). (E) Il23r expression by cLPLs as evaluated by qRT-PCR. Data are normalized on Hprt expression. (F and G) 129SvEv Rag2^−/− mice were infected with H. hepaticus for 8 wk and CD45⁺lin⁻Sca1⁺Thy1.2^Hi ILCs were FACS sorted from the colon. (F) ILCs were cultured overnight in complete medium alone (n/a) or in the presence of 10 ng/ml IL-1β. Il23r and Rorc expression levels were evaluated by qRT-PCR and normalized on Hprt expression. Data are shown as mean ± SEM from 2 pooled independent experiments. In each experiment, 10–18 mice were pooled. (G) ILCs were cultured overnight in complete medium alone (n/a) or in the presence of 10 ng/ml IL-23. Il1r1 expression was evaluated by qRT-PCR and normalized on Hprt expression. Data are shown as mean ± SEM from two pooled independent experiments. In each experiment, 10–18 mice were pooled. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.