Tissue factor–bearing exosome secretion from human mechanically stimulated bronchial epithelial cells in vitro and in vivo

• Published in: Journal of allergy and clinical immunology Vol. 130; no. 6; pp. 1375 - 1383
• Main Authors: Park, Jin-Ah, Sharif, Asma S., Tschumperlin, Daniel J., Lau, Laurie, Limbrey, Rachel, Howarth, Peter, Drazen, Jeffrey M.
• Format: Journal Article
• Language: English
• Published: New York, NY Mosby, Inc 01.12.2012; Elsevier; Elsevier Limited
• Subjects: Adult; Age; Aged; Airway management; Airway Remodeling; Allergy and Immunology; Angiogenesis; Asthma; Asthma - immunology; Asthma - pathology; Atmospheric pressure; Biological and medical sciences; Biopsy; blood coagulation; Bronchi - immunology; Bronchi - pathology; bronchial epithelium; Bronchoalveolar Lavage Fluid - immunology; bronchoconstriction; Bronchoconstriction - immunology; Cells, Cultured; Cellular Microenvironment - immunology; Chronic obstructive pulmonary disease, asthma; epithelial cells; Epithelial Cells - immunology; epithelium; exosomes; Exosomes - immunology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; gene expression regulation; Humans; immunohistochemistry; Immunopathology; in vivo studies; Kinases; mechanotransduction; Mechanotransduction, Cellular; Medical sciences; messenger RNA; Microscopy; Middle Aged; patients; Pneumology; Proteins; Respiratory distress syndrome; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; secretion; Thromboplastin - genetics; Thromboplastin - metabolism; tissue factor; ultracentrifugation; water; Young Adult; NHBECs; tissue factor; mechanotransduction; BALF; PKC; CM; bronchoconstriction; Bis 1; EBBs; Asthma; TF; exosomes; bronchial epithelium; GAPDH; TEM; ALI; Protein kinase C; Bisindoylmaleimide 1; Normal human bronchial epithelial cells; Conditioned media; Air-liquid interface; Glyceraldehyde-3-phosphate; Transmission electron microscopy; Bronchoalveolar lavage fluid; Endobronchial biopsies; Human; Lung disease; Immunopathology; Respiratory disease; Secretion; Tissue factor; Bronchus; Epithelium; Respiratory system; In vitro; Respiratory tract; In vivo; Bronchoconstriction; Immunology; Exosome; Bronchus disease; Epithelial cell; Obstructive pulmonary disease
• ISSN: 0091-6749; 1097-6825; 1097-6825
• DOI: 10.1016/j.jaci.2012.05.031

Tissue factor (TF), a primary initiator of blood coagulation, also plays a pivotal role in angiogenesis. TF expression in the airways is associated with asthma, a disease characterized in part by subepithelial angiogenesis. To determine potential sources of TF and the mechanisms of its availability in the lung microenvironment. Normal human bronchial epithelial cells grown in air-liquid interface culture were subjected to a compressive stress of 30 cm H2O; this is comparable to that generated in the airway epithelium during bronchoconstriction in asthma. Conditioned media and cells were harvested to measure TF mRNA and TF protein. We also tested bronchoalveolar lavage fluid and airway biopsies from asthmatic patients and healthy controls for TF. TF mRNA was upregulated 2.2-fold after 3 hours of stress compared with unstressed cells. Intracellular and secreted TF proteins were enhanced 1.6-fold and more than 50-fold, respectively, compared with those of control cells after the onset of compression. The amount of TF in the bronchoalveolar lavage fluid from patients with asthma was found at mean concentrations that were 5 times greater than those of healthy controls. Immunohistochemical staining of endobronchial biopsies identified epithelial localization of TF with increased expression in asthma. Exosomes isolated from the conditioned media of normal human bronchial epithelial cells and the bronchoalveolar lavage fluid of asthmatic subjects by ultracentrifugation contained TF. Our in vitro and in vivo studies show that mechanically stressed bronchial epithelial cells are a source of secreted TF and that exosomes are potentially a key carrier of the TF signal.