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. 2017 Mar 6;214(3):669–680. doi: 10.1084/jem.20161435

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Figure 5. — Increased functional IL-4Rα expression in STAT3^mut and ERBB2IP^mut naive lymphocytes promotes TGF-β–dependent GATA3 expression in CD4 T cells. (A) Contour plots demonstrating expression of IL-4Rα ex vivo on naive (CD27⁻) B cells (top) and contour plots showing CD27 and IL-4Rα coexpression and gating strategy of naive (CD45RO⁻) T cells (bottom) from control, STAT3^mut, and ERBB2IP^mut patients. (B) Combined ex vivo IL-4Rα expression on naive lymphocytes from control (n = 19), STAT3^mut (n = 17), and ERBB2IP^mut (n = 3) individuals. MFI on naive (CD27⁻) B cells (left) and percentage of naive (CD27⁻) B cells (middle) and of CD27⁺ naive (CD45RO⁻) CD4 cells (right) expressing IL-4Rα are shown. (C) Percentage of naive (CD27⁻) B cells from controls (n = 8), STAT3^mut (n = 8), and ERBB2IP^mut (n = 3) individuals expressing the STAT6-target CD23 ex vivo or after short-term stimulation with IL-4 as indicated. (D) IL-4Rα and resulting GATA3 expression in control, STAT3^mut, and ERBB2IP^mut naive CD4 T cells after activation (CD69⁺) with weak TCR stimulation under nonskewing conditions in the presence or absence of a TGF-β1–neutralizing antibody (αTGFβ1), a TGFβR1 inhibitor (TGFβR1i), or a selective SMAD3 inhibitor (SMAD3i). Contour plots (left) and combined GATA3 MFI (right) are shown. (E) Comparison of GATA3 expression in activated (CD69⁺) control naive CD4 cells, as normalized MFI, cultured with 10 µg/ml neutralizing IL-4 antibody (αIL-4) or SMAD3i under the same conditions as in D. Data are representative or combined from at least three independent experiments and are represented as the mean ± SEM. Unpaired two-tailed Student’s t tests and Mann-Whitney tests were used where appropriate. *, P < 0.05; **, P < 0.005; ***, P < 0.001.