Expression of IL-2 by B cells drives a distinct ILC2-eosinophil–oriented cellular circuit. (A) ELISA of IL-2 cytokine in serum. n = 8–10. (B) ELISA of IL-2 expression in splenic tissue. n = 6–12. (C) Spleen weight and cellularity of 4–6-wk-old Cd19creRosaIL-2 and littermate controls. n = 9–15. (D) Representative tSNE of high-parameter flow cytometry data from splenic viable cells and average frequency of each cluster per mouse. (E) Representative gating and frequency of eosinophils in spleen from WT, CD4creRosaIL-2, Cd19creRosaIL-2, or WT mice transferred with CD4creRosaIL-2 CD4+ or CD8+ T cells. n = 6–25. SSC, side scatter. (F) Luminex analysis of indicated cytokine in serum. n = 5–15. (G) IL-5 expression among cell lineages in spleen. n = 6–9. pDC, plasmacytoid DC. (H) Frequency of GATA3+ ILC2 among total ILCs in spleen. n = 4–6. (I) Frequency of ILC2 in spleen. n = 4–6. (J) pSTAT5 in freshly isolated ILC2 cells. n = 4. MFI, mean fluorescence intensity. (K) CD25 MFI of ILC2 in spleen. n = 4. (L) Frequency of splenic eosinophils in mice treated with anti−IL-5 neutralizing antibody. n = 3–8. (M) Survival analysis. n = 7–9. (N) Representative immunofluorescence staining of pSTAT5, CD3, and B220 in spleen (scale bars: 500 μm; 15 μm for inset). n = 5–6. (O) Frequency of pSTAT5+ cells found in splenic B cell zones by immunofluorescence. n = 5–6. Data pooled from (A–E and G–O) or representative of (F) at least two independent experiments. Significance was tested by one-way ANOVA (A, B, E, F, and L), unpaired t test (C and H–K), two-sample Kolmogorov–Smirnov test (D), Mantel–Cox log-rank test (M), or two-way ANOVA (O).
Figure 8.

Expression of IL-2 by B cells drives a distinct ILC2-eosinophil–oriented cellular circuit. (A) ELISA of IL-2 cytokine in serum. n = 8–10. (B) ELISA of IL-2 expression in splenic tissue. n = 6–12. (C) Spleen weight and cellularity of 4–6-wk-old Cd19creRosaIL-2 and littermate controls. n = 9–15. (D) Representative tSNE of high-parameter flow cytometry data from splenic viable cells and average frequency of each cluster per mouse. (E) Representative gating and frequency of eosinophils in spleen from WT, CD4creRosaIL-2, Cd19creRosaIL-2, or WT mice transferred with CD4creRosaIL-2 CD4+ or CD8+ T cells. n = 6–25. SSC, side scatter. (F) Luminex analysis of indicated cytokine in serum. n = 5–15. (G) IL-5 expression among cell lineages in spleen. n = 6–9. pDC, plasmacytoid DC. (H) Frequency of GATA3+ ILC2 among total ILCs in spleen. n = 4–6. (I) Frequency of ILC2 in spleen. n = 4–6. (J) pSTAT5 in freshly isolated ILC2 cells. n = 4. MFI, mean fluorescence intensity. (K) CD25 MFI of ILC2 in spleen. n = 4. (L) Frequency of splenic eosinophils in mice treated with anti−IL-5 neutralizing antibody. n = 3–8. (M) Survival analysis. n = 7–9. (N) Representative immunofluorescence staining of pSTAT5, CD3, and B220 in spleen (scale bars: 500 μm; 15 μm for inset). n = 5–6. (O) Frequency of pSTAT5+ cells found in splenic B cell zones by immunofluorescence. n = 5–6. Data pooled from (A–E and G–O) or representative of (F) at least two independent experiments. Significance was tested by one-way ANOVA (A, B, E, F, and L), unpaired t test (C and H–K), two-sample Kolmogorov–Smirnov test (D), Mantel–Cox log-rank test (M), or two-way ANOVA (O).

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