Polyinosinic:polycytidylic acid induces protein kinase D–dependent disassembly of apical junctions and barrier dysfunction in airway epithelial cells - 30/11/11
, Steve N. Georas, MD b, ⁎ 
Corresponding author: Steve N. Georas, MD, Division of Pulmonary and Critical Care Medicine, University of Rochester Medical Center, 601 Elmwood Ave, Rochester, NY, 14610.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.
Objectives |
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.
Le texte complet de cet article est disponible en PDF.Key words : Asthma, polyI:C, Toll-like receptor 3, epithelial permeability, protein kinase C, tight junctions, adherens junctions
Abbreviations used : AJ, AJC, dsRNA, HBSS, LDH, MDA5, MLCK, NHBE, NM II, pAb, PKC, PKD, polyI:C, PRR, RIG-I, ROCK, siRNA, TEER, TJ, TLR, ZO-1
Plan
| Supported by National Institutes of Health (NIH) grant R01 HL071933 (to S.N.G.), Pilot Project Funding supported by NIH grant P30 ES 01247 (to S.N.G.), NIH grants R01 DK083968 and DK084953 (to A.I.I.), NIH grant T32 HD057821 (to F.R.), NIH grant T32 HL066988 (to T.J.C.), the National Eczema Association (to A.D.), and a Bradford Fellowship from the University of Rochester Department of Pediatrics (to F.R.). |
|
| Disclosure of potential conflict of interest: L. A. Beck receives research support from Centocor, Regeneron, and Genentech. S. N. Georas receives research support from the National Institutes of Health. The rest of the authors declare that they have no relevant conflicts of interest. |
Vol 128 - N° 6
P. 1216 - décembre 2011 Retour au numéro
Article précédent
- Inflammation-mediated upregulation of centrosomal protein 110, a negative modulator of ciliogenesis, in patients with chronic rhinosinusitis
- Yinyan Lai, Bei Chen, Jianbo Shi, James N. Palmer, David W. Kennedy, Noam A. Cohen
- Plasticity of airway epithelial cells
- Pedro C. Avila
Bienvenue sur EM-consulte, la référence des professionnels de santé.
L’accès au texte intégral de cet article nécessite un abonnement.
Déjà abonné à cette revue ?