title: Polyinosinic:polycytidylic acid induces protein kinase D-dependent disassembly of apical junctions and barrier dysfunction in airway epithelial cells.
abstract: BACKGROUND
  Disruption of the epithelial barrier might be a risk factor for allergen sensitization and asthma. Viral respiratory tract infections are strongly associated with asthma exacerbation, but the effects of respiratory viruses on airway epithelial barrier function are not well understood. Many viruses generate double-stranded RNA, which can lead to airway inflammation and initiate an antiviral immune response.
  
  
  OBJECTIVE
  We investigated the effects of the synthetic double-stranded RNA polyinosinic:polycytidylic acid (polyI:C) on the structure and function of the airway epithelial barrier in vitro.
  
  
  METHODS
  16HBE14o- human bronchial epithelial cells and primary airway epithelial cells at an air-liquid interface were grown to confluence on Transwell inserts and exposed to polyI:C. We studied epithelial barrier function by measuring transepithelial electrical resistance and paracellular flux of fluorescent markers and structure of epithelial apical junctions by means of immunofluorescence microscopy.
  
  
  RESULTS
  PolyI:C induced a profound decrease in transepithelial electrical resistance and increase in paracellular permeability. Immunofluorescence microscopy revealed markedly reduced junctional localization of zonula occludens-1, occludin, E-cadherin, β-catenin, and disorganization of junction-associated actin filaments. PolyI:C induced protein kinase D (PKD) phosphorylation, and a PKD antagonist attenuated polyI:C-induced disassembly of apical junctions and barrier dysfunction.
  
  
  CONCLUSIONS
  PolyI:C has a powerful and previously unsuspected disruptive effect on the airway epithelial barrier. PolyI:C-dependent barrier disruption is mediated by disassembly of epithelial apical junctions, which is dependent on PKD signaling. These findings suggest a new mechanism potentially underlying the associations between viral respiratory tract infections, airway inflammation, and allergen sensitization.
authors: Division of Pediatric Pulmonary, Department of Pediatrics, University of Rochester Medical Center, Rochester, NY 14610, USA.
authors: Fariba
authors: F
authors: Rezaee
authors: Nida
authors: N
authors: Meednu
authors: Jason A
authors: JA
authors: Emo
authors: Bahman
authors: B
authors: Saatian
authors: Timothy J
authors: TJ
authors: Chapman
authors: Nayden G
authors: NG
authors: Naydenov
authors: Anna
authors: A
authors: De Benedetto
authors: Lisa A
authors: LA
authors: Beck
authors: Andrei I
authors: AI
authors: Ivanov
authors: Steve N
authors: SN
authors: Georas
mesh_terms: D001249:Asthma / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D015151:Immunoblotting; D007369:Interferon Inducers / Q000276:immunology / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010539:Permeability; D011070:Poly I-C / Q000276:immunology* / Q000378:metabolism; D011493:Protein Kinase C / Q000276:immunology / Q000378:metabolism*; D034622:RNA Interference; D020545:Respiratory Mucosa / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D012141:Respiratory Tract Infections / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction / Q000276:immunology*; D019108:Tight Junctions / Q000276:immunology / Q000378:metabolism / Q000473:pathology
chemical_list: D007369:Interferon Inducers; C089212:protein kinase D; D011493:Protein Kinase C; D011070:Poly I-C