Figure 6.
Blimp-1–deficient ILC2s are poor type 2 cytokine producers in vitro and provoke less eosinophilia during allergic lung inflammation. (A) Concentrations of the indicated cytokine as measured in the supernatant 3 days after stimulation of sort-purified small intestinal ILC2s of Ctrl (Prdm1fl/fl) and cKO (Nmur1CrePrdm1fl/fl) mice with IL-7, IL-25, and IL-33 (n = 5 mice per group). (B) Representative flow cytometry plots showing lung ILC2s in papain-treated Ctrl and cKO mice. (C) Quantification of lung ILC2s (from B) in papain-treated Ctrl and cKO mice. (D) Representative flow cytometry plots showing lung and bronchoalveolar lavage (BAL) eosinophils in papain-treated Ctrl and cKO mice. (E) Quantification and counts of eosinophils (from D) in papain-treated Ctrl and cKO mice. (F and G) Flow cytometric analysis and quantification of IL-5– and IL-13–expressing lung ILC2s in papain-treated Ctrl and cKO mice. (B–G) Data are representative of two independent experiments; n = 4–5 mice per group. Student’s t test; *P < 0.05; **P < 0.01; ***P < 0.001. Refer to the image caption for details. Panel A shows cytokine measurements from stimulated small intestinal ILC2s show reduced IL-5, IL-13, and IL-9 production in Blimp-1–deficient cells compared with controls, indicating impaired type 2 effector function. Panel B shows flow cytometry plots of lung ILC2s after papain challenge show a reduced ILC2 population in cKO mice compared with control mice. Panel C shows quantification confirms decreased frequency of lung ILC2s in papain-treated cKO mice relative to controls. Panel D shows representative flow cytometry plots of lung and BAL eosinophils show reduced eosinophil accumulation in Blimp-1–deficient mice following papain treatment. Panel E shows that quantitative analysis demonstrates significantly lower percentages and total numbers of eosinophils in lung and BAL of cKO mice. Panel F shows flow cytometric analysis and intracellular staining shows fewer IL-5 plus and IL-13 plus lung ILC2s in papain-treated cKO mice compared with controls. Panel G shows quantification confirms reduced IL-5 plus IL-13 plus ILC2 frequencies in Blimp-1–deficient mice, indicating weakened type 2 inflammatory responses.

Blimp-1–deficient ILC2s are poor type 2 cytokine producers in vitro and provoke less eosinophilia during allergic lung inflammation. (A) Concentrations of the indicated cytokine as measured in the supernatant 3 days after stimulation of sort-purified small intestinal ILC2s of Ctrl (Prdm1fl/fl) and cKO (Nmur1CrePrdm1fl/fl) mice with IL-7, IL-25, and IL-33 (n = 5 mice per group). (B) Representative flow cytometry plots showing lung ILC2s in papain-treated Ctrl and cKO mice. (C) Quantification of lung ILC2s (from B) in papain-treated Ctrl and cKO mice. (D) Representative flow cytometry plots showing lung and bronchoalveolar lavage (BAL) eosinophils in papain-treated Ctrl and cKO mice. (E) Quantification and counts of eosinophils (from D) in papain-treated Ctrl and cKO mice. (F and G) Flow cytometric analysis and quantification of IL-5– and IL-13–expressing lung ILC2s in papain-treated Ctrl and cKO mice. (B–G) Data are representative of two independent experiments; n = 4–5 mice per group. Student’s t test; *P < 0.05; **P < 0.01; ***P < 0.001.

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