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Epithelial permeability alterations in an in vitro air–liquid interface model of allergic fungal rhinosinusitis

Authors

  • Kyle A. Den Beste BS,

    1. Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA
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  • Elizabeth K. Hoddeson MD,

    1. Department of Otolaryngology–Head and Neck Surgery, Emory University, Atlanta, GA
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  • Charles A. Parkos MD, PhD,

    1. Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA
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  • Asma Nusrat MD,

    1. Epithelial Pathobiology and Mucosal Inflammation Research Unit, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA
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  • Sarah K. Wise MD

    Corresponding author
    1. Department of Otolaryngology–Head and Neck Surgery, Emory University, Atlanta, GA
    • Department of Otolaryngology–Head and Neck Surgery, Emory University, 550 Peachtree Street, MOT 9th Floor, Atlanta, GA 30308
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  • Funding sources for the study: Short Term Training in Health Professional Schools, National Institutes of Health (T35-HL007473 to K.A.D., Principal Investigator - Robinna G. Lorenz, MD, PhD); American Rhinologic Society New Investigator Award (S.K.W.); Clinical and Translational Science Award Program, National Institutes of Health, National Center for Research Resources (KL2 RR025009 and UL1 RR025008, both to S.K.W., Principal Investigator - David Stephens, MD); Structure function studies in intestinal epithelial JAM, National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases (DK061379 to C.A.P.); Neutrophil interactions with intestinal epithelial cells, National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases (DK072564 to C.A.P.); Intestinal epithelial tight junction structure-function, National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases (DK059888 to A.N.). Potential conflict of interest: None provided.

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 T helper 2 (Th2) cytokines and antigen-specific immunoglobulin E (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 among cultured primary sinonasal cells from AFRS patients vs noninflammatory controls.

Methods:

Epithelial cells isolated from paranasal sinus mucosa of AFRS and noninflammatory control patients were grown to confluence on permeable supports and transitioned to air–liquid interface (ALI). Transepithelial 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.

Conclusion:

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. © 2013 ARS–AAOA, LLC.

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