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Impaired barrier function in patients with house dust mite–induced allergic rhinitis is accompanied by decreased occludin and zonula occludens-1 expression 

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Presentation on theme: "Impaired barrier function in patients with house dust mite–induced allergic rhinitis is accompanied by decreased occludin and zonula occludens-1 expression "— Presentation transcript:

1 Impaired barrier function in patients with house dust mite–induced allergic rhinitis is accompanied by decreased occludin and zonula occludens-1 expression  Brecht Steelant, MSc, Ricard Farré, PhD, Paulina Wawrzyniak, MSc, Jochen Belmans, MSc, Emily Dekimpe, MSc, Hanne Vanheel, PhD, Laura Van Gerven, MD, PhD, Inge Kortekaas Krohn, MSc, Dominique M.A. Bullens, MD, PhD, Jan L. Ceuppens, MD, PhD, Cezmi A. Akdis, MD, Guy Boeckxstaens, MD, PhD, Sven F. Seys, PhD, Peter W. Hellings, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 137, Issue 4, Pages e5 (April 2016) DOI: /j.jaci Copyright © 2016 The Authors Terms and Conditions

2 Fig 1 Impaired mucosal barrier function ex vivo in patients with HDM-induced AR. Nasal biopsy specimens were isolated from control subjects and patients with HDM-induced AR for evaluation of mucosal integrity in Ussing chambers. A, Transtissue resistance measured with Ussing chambers. B, FD4 epithelial permeability. C, Transtissue resistance negatively correlated with FD4 passage in control subjects and patients with HDM-induced AR. D-E, Lower mRNA expression of occludin and ZO-1 in patients with HDM-induced AR compared with that in control subjects. Relative mRNA expression versus the housekeeping genes encoding β-actin and β2-microglobulin is shown. F, Transtissue resistance negatively correlated with visual analog scale scores for major symptoms in patients with HDM-induced AR. G, Representative immunofluorescence staining for occludin and ZO-1 in 8 control subjects and 9 patients with HDM-induced AR. *P < .05. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

3 Fig 2 Impaired NEC function in vitro in patients with HDM-induced AR. NECs were isolated from the inferior turbinate and cultured in vitro. A, TER in ALI cultures after 21 days in patients with HDM-induced AR and control subjects. B, FD4 passage in patients with HDM-induced AR and control subjects. C, TER negatively correlated with FD4 passage in control subjects and patients with HDM-induced AR. D-E, Lower mRNA expression of occludin and ZO-1 in patients with HDM-induced AR compared with control subjects. Relative mRNA expression versus the housekeeping genes encoding β-actin and β2-microglobulin is shown. F, Representative immunofluorescence staining showed less clear arrangement for occludin and ZO-1 in patients with HDM-induced AR compared with that in control subjects. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

4 Fig 3 INSs restored barrier function ex vivo. Nasal biopsy specimens were isolated from patients with HDM-induced AR using INSs (n = 6) and not using INSs (n = 9) for evaluation of mucosal integrity in Ussing chambers. A, Transtissue resistance measured with Ussing chambers. B, FD4 epithelial permeability. C-D, mRNA expression of occludin and ZO-1 was increased in patients with HDM-induced AR using INSs. Relative mRNA expression versus the housekeeping genes encoding β-actin and β2-microglobulin is shown. E, Representative immunofluorescence staining showed a restored organization for occludin and ZO-1 in 3 patients with HDM-induced AR using INSs compared with 6 patients with HDM-induced AR not using INSs. The data obtained for control subjects and patients with HDM-induced AR using INSs are duplicated from Fig 1. *P < .05. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

5 Fig 4 Regulation of epithelial barrier function in vitro by IL-4, IFN-γ, and FP in healthy control subjects. NECs were isolated from the inferior turbinates of control donors (n = 5) and cultured in vitro to evaluate the effect of proinflammatory cytokines and FP on NEC integrity. A, Effect of IL-4, FP, and IL-4 plus FP on epithelial integrity, as measured with TER in duplicate. B, Effect of IL-4, FP, and IL-4 plus FP on FD4 epithelial permeability. C, IFN-γ did not influence TER measurements in control subjects. D, No effect of IFN-γ on FD4 epithelial permeability in control subjects. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

6 Fig 5 RU-486 antagonized the effect of FP on epithelial integrity. NECs were isolated from the inferior turbinates of control subjects (n = 5) and cultured in vitro to study the effect of FP and its antagonist, RU-486. Relative TER changes in NEC cultures after stimulation. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

7 Fig 6 HDM-induced murine model of allergic airway disease to study barrier function in vivo. A, Schematic representation of the murine model. e.n., Endonasal. B, FD4 passage in the serum at different time points. Data are presented as means ± SEMs. *P < .05 and ***P < .001. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

8 Fig E1 Regulation of epithelial barrier function in vitro by IL-4, IFN-γ, and FP in patients with HDM-induced AR. NECs were isolated from the inferior turbinates of patients with HDM-induced AR (n = 5) and cultured in vitro to evaluate the effect of proinflammatory cytokines and FP on NEC integrity. A, Effect of IL-4, FP, and IL-4 plus FP on epithelial integrity, as measured with TER in duplicate. B, Effect of IL-4, FP, and IL-4 plus FP on FD4 epithelial permeability. C-D, IFN-γ did not affect TER and FD4 measurements. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

9 Fig E2 FP upregulates mRNA expression of occludin and ZO-1 in primary NEC cultures from healthy control subjects (n = 5) when stimulated with FP for 72 hours. FP increased mRNA expression of occludin (A) and ZO-1 (B) and had no effect on claudin-1 (C) and claudin-4 (D) expression. Relative mRNA expression versus that of the housekeeping genes encoding β-actin and β2-microglobulin. Data are presented as medians and interquartile ranges. *P < .05. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

10 Fig E3 Differential cell counts in bronchoalveolar lavage fluid in HDM-challenged mice. Eosinophilic bronchoalveolar lavage counts were significantly lower 1 and 24 hours after the last challenge between sham/HDM-challenged and FP/HDM-challenged mice. After 72 hours, no difference was found in eosinophilic cell counts. *P < .05 and **P < .01, Kruskal-Wallis test with post hoc analysis. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

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