Increased density of intraepithelial mast cells in patients with exercise-induced bronchoconstriction regulated through epithelially derived thymic stromal.

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Increased density of intraepithelial mast cells in patients with exercise-induced bronchoconstriction regulated through epithelially derived thymic stromal lymphopoietin and IL-33 Ying Lai, PhD, William A. Altemeier, MD, John Vandree, MD, Adrian M. Piliponsky, PhD, Brian Johnson, BS, Cara L. Appel, BS, Charles W. Frevert, PhD, Dallas M. Hyde, PhD, Steven F. Ziegler, PhD, Dirk E. Smith, PhD, William R. Henderson, MD, Michael H. Gelb, PhD, Teal S. Hallstrand, MD, MPH Journal of Allergy and Clinical Immunology Volume 133, Issue 5, Pages 1448-1455 (May 2014) DOI: 10.1016/j.jaci.2013.08.052 Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Airway gene expressions of specific mast cell proteases are increased in a precise phenotype of asthma. Quantitative PCR of airway epithelial brushings (A-C) and induced sputum cells (D-F) for TPSAB1, CPA3, and CMA1 expression is shown. The overall P value for the Kruskal-Wallis test is shown in the upper left of each pane, and the P values for post hoc tests are shown above the horizontal bars. NS, Not significant. Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Quantification of the density of intraepithelial mast cells (MCs) by using design-based stereology. The density of intraepithelial mast cells was quantified based on the numeric density of intraepithelial mast cells relative to the volume of the airway epithelium (Nv MC, epi; A) and the numeric density of mast cells relative to the surface area of the basal lamina (Ns MC, bala; B). NS, Not significant. Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 A mechanical scratch wound initiates TSLP release from primary airway epithelial cells in organotypic culture. Airway epithelial cells at passage 2 from asthmatic and control subjects differentiated in organotypic culture release TSLP protein into the basolateral media after a series of mechanical scratch wounds. The release of TSLP was greater from epithelial cells derived from asthmatic patients. BL, Baseline. Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Osmotic stress in murine lung tissue initiates the release of biologically active IL-33. Conditioned medium from explanted lung tissue exposed to osmotic stress in the form of sorbitol (A) or mannitol (B) increased ST2 reporter activity in Ba/F3 cells that was related to the amount of osmotic stress. †Test for linear trend. Release of ST2 activity was similar for equal molar concentrations of sorbitol or mannitol (C). Allergen challenge increased the basal release of ST2-activating activity and increased the release of this activity in response to osmotic stress (D). Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Influence of IL-33 and TSLP on IgE-dependent mast cell production of CysLTs and PGD2. Treatment of CBMCs for 7 days with IL-33 in combination with TSLP increased CysLT formation in response to the FcεRI-activating antibody CRA1 both without passive sensitization (A) and after passive sensitization with polyclonal IgE for 7 days (B). There was no increase in PGD2 formation in response to IL-33, TSLP, or both without passive sensitization (C) or after passive sensitization (D). NS, Not significant. Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Effects of IL-25 and IL-13 on IgE-dependent mast cell CysLT and PGD2 formation. Treatment of mature CBMCs with IL-25 did not alter the formation of IgE receptor-mediated (CRA1) formation of PGD2 (A) or CysLTs (B) with or without passive sensitization. Treatment of CBMCs with IL-13 did not alter the formation of PGD2 either with or without passive sensitization (C). The formation of CysLTs was increased by IL-13 after passive sensitization but not without passive sensitization of the CBMCs (D). NS, Not significant. Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E5 Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E6 Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E7 Journal of Allergy and Clinical Immunology 2014 133, 1448-1455DOI: (10.1016/j.jaci.2013.08.052) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions