An ex vivo model of severe asthma using reconstituted human bronchial epithelium Delphine Gras, PhD, Arnaud Bourdin, MD, PhD, Isabelle Vachier, PhD, Laure.

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An ex vivo model of severe asthma using reconstituted human bronchial epithelium Delphine Gras, PhD, Arnaud Bourdin, MD, PhD, Isabelle Vachier, PhD, Laure de Senneville, Caroline Bonnans, PhD, Pascal Chanez, MD, PhD Journal of Allergy and Clinical Immunology Volume 129, Issue 5, Pages 1259-1266.e1 (May 2012) DOI: 10.1016/j.jaci.2012.01.073 Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 RBM and epithelial thickness and goblet cell numbers were increased in patients with severe asthma. A and B, RBM (Fig 1, A) and epithelial thickness (Fig 1, B), expressed in micrometers, were measured in endobronchial biopsy specimens from control subjects and patients with mild or severe asthma. C, The number of goblet cells, expressed as a percentage of positive cells, was obtained by using periodic acid–Schiff (PAS) staining. Results are expressed as medians and interquartile ranges with 10th and 90th percentiles. *P < .05, Kruskall-Wallis followed by post hoc test. Journal of Allergy and Clinical Immunology 2012 129, 1259-1266.e1DOI: (10.1016/j.jaci.2012.01.073) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Morphologic analysis of bronchial epithelia in air-liquid interface cultures. A and B, Representative transmission electron microscopy of fully differentiated epithelium from patients with severe (Fig 2, A) and mild (Fig 2, B) asthma (original magnification ×4000). BC, Basal cell; CC, ciliated cell; GC, goblet cell. C, Goblet cell containing electron-lucent secretory granules (original magnification ×12,000). D, Apical surface of the epithelium showing cilia (C) and tight junction (TJ; original magnification ×12,000). E and F, Longitudinal sections (Fig 2, E) and cross-sections (Fig 2, F) of cilia showing microtubular doublets (MT; original magnification ×50,000). Journal of Allergy and Clinical Immunology 2012 129, 1259-1266.e1DOI: (10.1016/j.jaci.2012.01.073) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Mucin secretion and gene expression in air-liquid interface cultures of bronchial epithelium from patients with asthma. A, Analysis of total mucin secretion by using ELLA. B and C, Semiquantitative analysis of MUC5AC (Fig 3, B) and MUC5B (Fig 3, C) mRNA, with β-actin as a control. Results are expressed as medians and interquartile ranges with 10th and 90th percentiles. *P < .05, Mann-Whitney U test. Journal of Allergy and Clinical Immunology 2012 129, 1259-1266.e1DOI: (10.1016/j.jaci.2012.01.073) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 IL-8 and LXA4 levels in apical wash samples from air-liquid interface cultures. A, Quantitative analysis of IL-8 production in epithelia from control subjects, patients with mild asthma, and patients with severe asthma (in nanograms per milliliter). B, Quantitative analysis of LXA4 production in epithelia from control subjects, patients with mild asthma, and patients with severe asthma (in picograms per milliliter). Results are expressed as medians and interquartile ranges with 10th and 90th percentiles. *P < .05, Kruskall-Wallis test followed by post hoc test. Journal of Allergy and Clinical Immunology 2012 129, 1259-1266.e1DOI: (10.1016/j.jaci.2012.01.073) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Expression of genes that regulate lipoxin biosynthesis in air-liquid interface epithelial cell cultures. Semiquantitative analysis of 15LO1 (A), 15LO2 (B), and 5LO (C) mRNA is shown, with β-actin as a control. Results are expressed as medians and interquartile ranges with 10th and 90th percentiles. *P < .05, Mann-Whitney U test. Journal of Allergy and Clinical Immunology 2012 129, 1259-1266.e1DOI: (10.1016/j.jaci.2012.01.073) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions