Mast cells infiltrate the esophageal smooth muscle in patients with eosinophilic esophagitis, express TGF-β1, and increase esophageal smooth muscle contraction

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Mast cells infiltrate the esophageal smooth muscle in patients with eosinophilic esophagitis, express TGF-β1, and increase esophageal smooth muscle contraction Seema S. Aceves, MD, PhD, Diana Chen, BS, Robert O. Newbury, MD, Ranjan Dohil, MD, John F. Bastian, MD, David H. Broide, MB, ChB Journal of Allergy and Clinical Immunology Volume 126, Issue 6, Pages 1198-1204.e4 (December 2010) DOI: 10.1016/j.jaci.2010.08.050 Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Mast cells in the epithelium (EPI) and LP. A, MCT infiltrating the epithelium, LP, and SM in control subjects and patients with EE (magnification ×100). B and C, Tryptase (Fig 1, B)– and chymase (Fig 1, C)–positive cell numbers. Bars represent medians. D-F, Epithelial and LP MCT (Fig 1, D), LP MCC (Fig 1, E; magnification ×100), and LP MCTC (tryptase, green; chymase, red; overlap, yellow; magnification ×400; Fig 1, F) from patients with EE. Journal of Allergy and Clinical Immunology 2010 126, 1198-1204.e4DOI: (10.1016/j.jaci.2010.08.050) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Esophageal SM is infiltrated by MCT. A and B, Representative image of SM MCT in control subjects (Fig 2, A) and patients with EE (Fig 2, B). C and D, Quantitation of MCT and MCC (Fig 2, C) and eosinophil (Fig 2, D) numbers in SM. Bars represent medians. Journal of Allergy and Clinical Immunology 2010 126, 1198-1204.e4DOI: (10.1016/j.jaci.2010.08.050) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Mast cells express TGF-β1, which increases esophageal SM contraction. A-C, TGF-β1–positive cells (green, Fig 3, A) coexpress tryptase (red, Fig 3, C; overlap, yellow, Fig 3, B; magnification ×200). D and E, Representative image (Fig 3, D) and numbers (Fig 3, E) of TGF-β1–positive cells (magnification ×400) in SM from patients with EE and control subjects. Bars represent medians. F and G, TGF-β1 treatment of esophageal SM cells decreases collagen gel size. H, Collagen gel size after histamine (HA), TGF-β1, and actinomycin D (AD). Bars represent means ± SEs. ∗P < .05. Journal of Allergy and Clinical Immunology 2010 126, 1198-1204.e4DOI: (10.1016/j.jaci.2010.08.050) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Effects of topical corticosteroid therapy on MCT and MCC numbers in the epithelium (EPI) and LP. Epithelial (A-C) and LP (D-F) tryptase- and chymase-positive cells and eosinophils before (pre) and after (post) budesonide in responders (7 or fewer epithelial eosinophils per hpf after therapy) and nonresponders (>20 epithelial eosinophils per hpf after therapy). Bars represent medians. Journal of Allergy and Clinical Immunology 2010 126, 1198-1204.e4DOI: (10.1016/j.jaci.2010.08.050) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Immunoblot for phosphorylated Smad3 using nuclear extracts from esophageal SM cells cultured in the absence (−) or presence (+) of TGF-β1 shows pSmad3 in the nucleus only after TGF-β1 treatment. Journal of Allergy and Clinical Immunology 2010 126, 1198-1204.e4DOI: (10.1016/j.jaci.2010.08.050) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions