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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: J Allergy Clin Immunol. 2019 Jun 5;144(4):1058–1073.e3. doi: 10.1016/j.jaci.2019.04.030

Figure 5. Type-II IL-4Rα signaling pathway is required for IL-13-induced SAP formation, luminal allergen translocation and induction of oral passive IgE- and active-food-induced anaphylaxis.

Figure 5.

(A) Diagram depicting experimental design for inducible deletion of IL-4Rα in intestinal secretory epithelial cells. (B) Immunofluorescence analysis for IL-4Rα (green) and MUC2 (white) in mouse SIs of IL-4RαWT and IL-4RαΔsecretory mice exposed to Rh-Dex. Green arrow depicts the cytoplasm of Rh-Dex+ MUC2+ GCs in IL-4RαWT mice that are IL-4Rα+. White arrow depicts cytoplasm of Rh-Dex+ MUC2+ GCs in IL-4RαΔsecretory that are IL-4Rα−. The dotted line indicates GCs. (C) Alcian blue staining with hematoxylin counterstain of SI from IL-4RαWT and IL-4RαΔsecretory mice following i.p. RU-486, (D) SI of IL9Tg IL-4RαWT and IL-9Tg IL-4RαΔsecretory mice treated with RU-486 and intraluminal SI challenge of Rh-Dex and OVA-TNP. Nucleus was visualized with DAPI and SAPs by Rh-Dex+ staining. (E) Δ Temperature (°C) (at 45 minutes) in IL9Tg IL-4RαWT and IL-9Tg IL-4RαΔsecretory mice treated with RU-486, IL-13 and TNP-IgE (described in A) following oral TNP-OVA challenge. *p < 0.05. (F) Experimental regime for neutralization of IL-13Rα1 using anti-IL-13Rα1 mAb in food allergic mice. Mice that demonstrated evidence of anaphylaxis (shock) following the 5th oral challenge were stratified into isotype control (iso control) or anti-IL-13Ra1 mAb treatment groups. Experimental analyses were performed following 7th oral challenge on Day 28. (G) Immunofluorescence analysis for MUC2 (green) and SAPs (Rh-Dex+), Alcian blue staining with hematoxylin counterstain of SI and Quantitation of antigen passing cells per villus/SI from isotype control (Iso Ctrl) of SI’s of control and anti-IL-13Rα1 treated mice. (H) ΔTemperature (°C), % of mice with diarrhea and serum MCPT-1 levels (at 45 minutes) in isotype control (Iso Ctrl) and anti-IL-13Rα1 treated mice following the 7th oral OVA challenge on day 28 indicated in (F). The fractions indicates the number of mice with diarrhea of the total number of mice. Scale bars are 50 μm (B, D, G, H) and 20 μm (C). i.v. intra-venous; i.p. intraperitoneal, O.g. oral gavage. * p < 0.05.