Epithelial Permeability Alterations in an In Vitro Air-Liquid Interface Model of Allergic Fungal Rhinosinusitis

• Type of Material: Article
• Authors: Kyle A. Den Beste, Emory University; Elizabeth K. Hoddeson, Emory University; Charles Parkos, Emory University; Asma Nusrat, Emory University; Sarah K. Wise, Emory University
• Language: English
• Date: 2013-01
• Publisher: Wiley: 12 months
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2013 American Rhinologic Society-American Academy of Otolaryngic Allergy, LLC
• Final Published Version (URL): http://onlinelibrary.wiley.com/doi/10.1002/alr.21077/abstract
• Title of Journal or Parent Work: International Forum of Allergy and Rhinology
• ISSN: 2042-6976
• Volume: 3
• Issue: 1
• Start Page: 19
• End Page: 25
• Grant/Funding Information: American Rhinologic Society New Investigator Award (S.K.W.); Short Term Training in Health Professional Schools, National Institutes of Health T35-HL007473 (K.A.D.); This work was supported in part by the following funding sources:; Structure function studies in intestinal epithelial JAM, National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases DK061379 (C.A.P.); Clinical and Translational Science Award Program, National Institutes of Health, National Center for Research Resources KL2 RR0025009 and UL1 RR025008 (S.K.W.); Neutrophil interactions with intestinal epithelial cells, National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases, DK072564 (C.A.P.); Intestinal epithelial tight junction structure-function, National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases, DK059888 (A.N.)
• Keywords: occludin; Adherens junction; epithelial permeability; allergic rhinitis; tight junction; E-cadherin; claudin-2; junction adhesion molecule-A; sinonasal epithelium; allergic fungal rhinosinusitis
• Research Categories: Health Sciences, Pathology

Abstract

Background Chronic rhinosinusitis (CRS) is an inflammatory upper-airway disease with numerous etiologies. Patients with a characteristic subtype of CRS, allergic fungal rhinosinusitis (AFRS), display increased expression of Th2 cytokines and antigen-specific IgE. Various sinonasal inflammatory conditions are associated with alterations in epithelial barrier function. The aim of this study was to compare epithelial permeability and intercellular junctional protein expression amongst cultured primary sinonasal cells from AFRS patients versus non-inflammatory controls. Methods Epithelial cells isolated from paranasal sinus mucosa of AFRS and non-inflammatory control patients were grown to confluence on permeable supports and transitioned to air-liquid interface (ALI). Trans-epithelial resistance (TER) was measured with a horizontal Ussing chamber to characterize the functional permeability of each cell type. After TER recordings were complete, a panel of intercellular junctional proteins was assessed by Western blot and immunofluorescence labeling followed by confocal microscopy. Results After 12 samples were measured from each group, we observed a 41% mean decrease in TER in AFRS cells (296±89 ohms × cm2) compared to control (503±134 ohms × cm2, P=0.006). TER deficits observed in AFRS were associated with decreased expression of the tight junction proteins occludin and Junctional Adhesion Molecule-A (JAM-A), and increased expression of a leaky tight junction protein claudin-2. Conclusions Cultured sinonasal epithelium from AFRS patients displayed increased epithelial permeability and altered expression of intercellular junctional proteins. Given that these cells were not incubated with inflammatory cytokines in vitro, the cultured AFRS epithelial alterations may represent a retained modification in protein expression from the in vivo phenotype.