IL-33-Induced IL-13 Production by Th2 Cells Is Dependent on a Signaling Complex between T1/ST2 and EGFR WT and Egfr fl/fl xCd4-cre (EGFR ΔCD4 ) mice were infected with H. polygyrus larvae, and on day 14 post infection, mLN were harvested. (A–C) Cells were stimulated with rIL-33, anti-CD3, or media in the presence of gefitinib, marimastat, or vehicle, and p-ERK (A and C) or IL-13 (B) expression was determined by intra-cellular staining and flow cytometry analysis. (D and E) Cells were stimulated with rHB-EGF, anti-CD3, or media, and IL-13 (D) and p-ERK (E) expression was determined by intra-cellular staining and flow cytometry analysis. (F) EGFR expression on stimulated mLN WT CD4 + T cells in the presence of monensin was analyzed by flow cytometry. (G) HEK293T cells were transfected as indicated with T1/ST2, the IL-1RacP, or the EGFR alone or in combination. Subsequently, the cells lysates were analyzed for the expression of the transfected proteins (input, left panel). The same lysates were also subjected to an EGFR-specific immunoprecipitation (EGFR-IP, right panel) or were treated with the isotype control (iso, right panel). Precipitates were analyzed by immunoblot. (H) mLN (upper) or HaCaT cells (lower) were stimulated with rIL-33, rHB-EGF, or media, and the EGFR phosphorylation at position Y1068 was determined by intra-cellular staining and flow cytometry analysis (upper) or immunoblot (lower). All data are representative of at least two independent experiments (mean ± SEM); results for individual mice are shown as dots. See also Figure S6 .
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