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The extra domain A of fibronectin is essential for allergen-induced airway fibrosis and hyperresponsiveness in mice  Martin Kohan, MSc, Andres F. Muro,

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Presentation on theme: "The extra domain A of fibronectin is essential for allergen-induced airway fibrosis and hyperresponsiveness in mice  Martin Kohan, MSc, Andres F. Muro,"— Presentation transcript:

1 The extra domain A of fibronectin is essential for allergen-induced airway fibrosis and hyperresponsiveness in mice  Martin Kohan, MSc, Andres F. Muro, PhD, Reem Bader, MSc, Neville Berkman, MBBCh, FRCP  Journal of Allergy and Clinical Immunology  Volume 127, Issue 2, Pages e5 (February 2011) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 EDA-FN is expressed in lung tissue and fibroblasts from OVA-treated mice. A and B, EDA-FN (Fig 1, A) and total fibronectin (Fig 1, B) mRNA in lungs of OVA-treated and control mice (n = 8-9). ∗P < .05. C, EDA-FN immunostaining of epithelium and subepithelial fibroblasts (arrows) in lung sections of OVA-treated mice. Representative images are shown; inserts indicate higher magnification. D, EDA-FN (red panels, light bars) and total fibronectin (green panels, dark bars) expression, calculated as integrated optical density (IOD), in lung primary fibroblasts (n = 4). ∗P < .05. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Reduced AHR in EDA−/− mice. A-C, Lung resistance (R; Fig 2, A), Newtonian resistance (Rn; Fig 2, B), and tissue damping (G; Fig 2, C) for increasing doses of methacholine. OVA-treated EDA−/− mice (gray circles, solid lines) showed reduced airway responsiveness in comparison with that seen in OVA-treated WT mice (black circles, solid lines; n = 8-9). ∗P < .05, ∗∗P < .01 (OVA-treated EDA−/− mice vs OVA-treated WT mice). Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Airway inflammation is unchanged in OVA-treated EDA−/− mice. A similar number of total cells are present in BAL fluid of WT and EDA−/− mice after 1 (A) and 5 (B) weeks of exposure to OVA. No differences in eosinophils number were observed after 1 (C) and 5 (D) weeks of OVA treatment. Data are presented as means ± SEMs (n = 8-9). Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Reduced airway fibrosis in EDA−/− mice. WT and EDA−/− mice were exposed to OVA and saline for 5 weeks. OVA-treated EDA−/− mice showed reduced lung collagen content (A) and peribronchial collagen deposition (B). No differences were observed in smooth muscle α-SMA–immunostained area (C and D) and airway mucus production (E and F). Data are presented as mean ± SEMs (n = 8-9). ∗∗P < .01. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Reduced expression of TGF-β1 and IL-13 in EDA−/− mice. Reduced lung tissue levels of TGF-β1 (A) and IL-13 (B) are seen in OVA-treated EDA−/− mice (n = 8-9). ∗∗∗P < Reduced release of TGF-β1 (C) and IL-13 (D) is seen in culture medium from OVA-treated EDA−/− lung fibroblasts after 24 hours of incubation (n = 4). ∗P < .05. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Impaired activation and differentiation of fibroblasts from lungs of OVA-treated EDA−/− mice. Lung fibroblasts from saline- and OVA-treated EDA−/− and WT mice were isolated as described in the printed Methods section and in the Methods section of this article's Online Repository. OVA-treated EDA−/− fibroblasts show reduced proliferation (A), migration (B), α-SMA expression (C), collagen deposition (D), and contractility (E; n = 4). ∗P < .05, ∗∗P < .01. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Schematic overview of study protocols for induction of acute (A) and chronic (B) allergen challenge.
Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 A, Lung sections from WT mice challenged with saline (Sal) or OVA were immunostained for EDA-FN (see Fig 1, C) and assigned an expression score based on the percentage of EDA-FN staining, as described in the Methods section of this article's Online Repository (n = 8-9 per group). ∗∗P < .01, Mann-Whitney test. B, Trichrome-stained lung sections from EDA−/− and WT mice exposed to saline or OVA (see Fig 4, B) were assigned a score based on the trichrome staining observed around the airways, as described in the Methods section of this article's Online Repository (n = 8-9 per group). ∗P < .05, WT OVA versus EDA−/− OVA, Kruskal-Wallis/Dunn test. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 No differences are observed in IL-4 (A) and GM-CSF (B) levels in lung homogenates from EDA−/− and WT mice exposed to OVA (n = 8-9 per group). Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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