T-helper 2 cytokines, transforming growth factor β1, and eosinophil products induce fibrogenesis and alter muscle motility in patients with eosinophilic esophagitis.
OBJECTIVE
Patients with eosinophilic esophagitis (EoE) often become dysphagic from the combination of organ fibrosis and motor abnormalities. We investigated mechanisms of dysphagia, assessing the response of human esophageal fibroblasts (HEFs), human esophageal muscle cells (HEMCs), and esophageal muscle strips to eosinophil-derived products.


METHODS
Biopsy specimens were collected via endoscopy from the upper, middle, and lower thirds of the esophagus of 18 patients with EoE and 21 individuals undergoing endoscopy for other reasons (controls). Primary cultures of esophageal fibroblasts and muscle cells were derived from 12 freshly resected human esophagectomy specimens. Eosinophil distribution was investigated by histologic analyses of full-thickness esophageal tissue. Active secretion of EoE-related mediators was assessed from medium underlying mucosal biopsy cultures. We quantified production of fibronectin and collagen I by HEF and HEMC in response to eosinophil products. We also measured the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 by, and adhesion of human eosinophils to, HEFs and HEMCs. Eosinophil products were tested in an esophageal muscle contraction assay.


RESULTS
Activated eosinophils were present in all esophageal layers. Significantly higher concentrations of eosinophil-related mediators were secreted spontaneously in mucosal biopsy specimens from patients with EoE than controls. Exposure of HEFs and HEMCs to increasing concentrations of eosinophil products or co-culture with eosinophils caused HEFs and HEMCs to increase secretion of fibronectin and collagen I; this was inhibited by blocking transforming growth factor β1 and p38 mitogen-activated protein kinase signaling. Eosinophil binding to HEFs and HEMCs increased after incubation of mesenchymal cells with eosinophil-derived products, and decreased after blockade of transforming growth factor β1 and p38 mitogen-activated protein kinase blockade. Eosinophil products reduced electrical field-induced contraction of esophageal muscle strips, but not acetylcholine-induced contraction.


CONCLUSIONS
In an analysis of tissues samples from patients with EoE, we linked the presence and activation state of eosinophils in EoE with altered fibrogenesis and motility of esophageal fibroblasts and muscle cells. This process might contribute to the development of dysphagia.
Department of Gastroenterology and Hepatology, Digestive Disease Institute, Cleveland, Ohio; Department of Pathobiology, Lerner Research Institute, Cleveland, Ohio.
Florian
F
Rieder
Department of Gastroenterology and Hepatology, Digestive Disease Institute, Cleveland, Ohio.
Ilche
I
Nonevski
Rhode Island Hospital and Brown University, Providence, Rhode Island.
Jie
J
Ma
Department of Gastroenterology and Hepatology, Digestive Disease Institute, Cleveland, Ohio.
Zhufeng
Z
Ouyang
Department of Pathobiology, Lerner Research Institute, Cleveland, Ohio.
Gail
G
West
Department of Biochemistry and Molecular Biology, Mayo Clinic Arizona, Scottsdale, Arizona.
Cheryl
C
Protheroe
Department of Laboratory Medicine/Pathology, Mayo Clinic Arizona, Scottsdale, Arizona.
Giovanni
G
DePetris
Department of Pathobiology, Lerner Research Institute, Cleveland, Ohio.
Anja
A
Schirbel
Department of Pathology, Cleveland Clinic Foundation, Cleveland, Ohio.
James
J
Lapinski
Department of Pathology, Cleveland Clinic Foundation, Cleveland, Ohio.
John
J
Goldblum
Division of Pediatric Pulmonology, Case Western Reserve University, Cleveland, Ohio.
Tracey
T
Bonfield
Department of Quantitative Health Sciences, Cleveland Clinic Foundation, Cleveland, Ohio.
Rocio
R
Lopez
Rhode Island Hospital and Brown University, Providence, Rhode Island.
Karen
K
Harnett
Department of Biochemistry and Molecular Biology, Mayo Clinic Arizona, Scottsdale, Arizona.
James
J
Lee
Division of Gastroenterology, Northwestern University Feinberg School of Medicine, Chicago, Illinois.
Ikuo
I
Hirano
Department of Gastroenterology and Hepatology, Digestive Disease Institute, Cleveland, Ohio.
Gary
G
Falk
Rhode Island Hospital and Brown University, Providence, Rhode Island.
Piero
P
Biancani
Department of Gastroenterology and Hepatology, Digestive Disease Institute, Cleveland, Ohio; Department of Pathobiology, Lerner Research Institute, Cleveland, Ohio. Electronic address: fiocchc@ccf.org.
Claudio
C
Fiocchi
D000368:Aged; D001706:Biopsy; D016022:Case-Control Studies; D002448:Cell Adhesion; D002450:Cell Communication; D002478:Cells, Cultured; D018920:Coculture Techniques; D024042:Collagen Type I / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism*; D003679:Deglutition; D003680:Deglutition Disorders / Q000209:etiology* / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D057765:Eosinophilic Esophagitis / Q000150:complications* / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D004804:Eosinophils / Q000276:immunology* / Q000378:metabolism; D004945:Esophagoscopy; D005260:Female; D005347:Fibroblasts / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D005353:Fibronectins / Q000378:metabolism; D005355:Fibrosis; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009092:Mucous Membrane / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D009119:Muscle Contraction; D032389:Myocytes, Smooth Muscle / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D018418:Th2 Cells / Q000276:immunology* / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D024042:Collagen Type I; D016207:Cytokines; D005353:Fibronectins; C573420:ICAM1 protein, human; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D048051:p38 Mitogen-Activated Protein Kinases
Immune Regulation; Primary Human Cells; Swallowing; Transmural Disease