Figure 7.
ADM potentiates ILC2-mediated lung inflammation. (A) Flow cytometric analysis of ILC2s stimulated in vitro with ADM, IL-33, or both in the presence of PMA and ionomycin for 2.5 h. (B) Percentage of IL-5+ and IL-13+ ILC2s as determined by intracellular cytokine staining. n = 3 sets/group. Data are representative of two independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by one-way ANOVA followed by Tukey’s test. (C) Experimental protocol for intranasal treatment with ADM. (D) Flow cytometric analysis (left) and the number (right) of eosinophils. (E) Total number of lung ILC2s and KLRG1+ ILC2s were assessed by flow cytometry. (F) Flow cytometric analysis of expression (left) and quantification of mean fluorescence intensity (MFI; right) of Sca-1 and KLRG1 in ILC2s. (G) Experimental protocol for N.b. infection treated with recombinant ADM. (H) Total numbers of ILC2s and KLRG1+ ILC2s were assessed on day 2 after N.b. infection. (I) Worm burden in the small intestine 7 d after infection; n = 3–5 mice/group. (D–I) Data are representative of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (Student’s t test). (J) The percentage of cytokine-positive ILC2s was quantified after in vitro culture with conditioned medium containing BAL fluid from WT and cKO mice in the absence or presence of ADM22-52. n = 2–3 mice/group pooled from two independent experiments. *, P < 0.05; **, P < 0.01 by one-way ANOVA followed by Tukey’s test. (K) Experimental protocol for intranasal administration of ADM22-52 and/or papain. (L and M) Total number of lung ILC2s, KLRG1+ ILC2s (L), and eosinophils (M) were assessed by flow cytometry. n = 3–4 mice/group pooled from two independent experiments; bar graphs show mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001 (Student’s t test).

ADM potentiates ILC2-mediated lung inflammation. (A) Flow cytometric analysis of ILC2s stimulated in vitro with ADM, IL-33, or both in the presence of PMA and ionomycin for 2.5 h. (B) Percentage of IL-5+ and IL-13+ ILC2s as determined by intracellular cytokine staining. n = 3 sets/group. Data are representative of two independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by one-way ANOVA followed by Tukey’s test. (C) Experimental protocol for intranasal treatment with ADM. (D) Flow cytometric analysis (left) and the number (right) of eosinophils. (E) Total number of lung ILC2s and KLRG1+ ILC2s were assessed by flow cytometry. (F) Flow cytometric analysis of expression (left) and quantification of mean fluorescence intensity (MFI; right) of Sca-1 and KLRG1 in ILC2s. (G) Experimental protocol for N.b. infection treated with recombinant ADM. (H) Total numbers of ILC2s and KLRG1+ ILC2s were assessed on day 2 after N.b. infection. (I) Worm burden in the small intestine 7 d after infection; n = 3–5 mice/group. (D–I) Data are representative of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (Student’s t test). (J) The percentage of cytokine-positive ILC2s was quantified after in vitro culture with conditioned medium containing BAL fluid from WT and cKO mice in the absence or presence of ADM22-52. n = 2–3 mice/group pooled from two independent experiments. *, P < 0.05; **, P < 0.01 by one-way ANOVA followed by Tukey’s test. (K) Experimental protocol for intranasal administration of ADM22-52 and/or papain. (L and M) Total number of lung ILC2s, KLRG1+ ILC2s (L), and eosinophils (M) were assessed by flow cytometry. n = 3–4 mice/group pooled from two independent experiments; bar graphs show mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001 (Student’s t test).

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