title: Rhinovirus disrupts the barrier function of polarized airway epithelial cells.
abstract: BACKGROUND
  Secondary bacterial infection following rhinovirus (RV) infection has been recognized in chronic obstructive pulmonary disease.
  
  
  OBJECTIVE
  We sought to understand mechanisms by which RV infection facilitates secondary bacterial infection.
  
  
  METHODS
  Primary human airway epithelial cells grown at air-liquid interface and human bronchial epithelial (16HBE14o-) cells grown as polarized monolayers were infected apically with RV. Transmigration of bacteria (nontypeable Haemophilus influenzae and others) was assessed by colony counting and transmission electron microscopy. Transepithelial resistance (R(T)) was measured by using a voltmeter. The distribution of zona occludins (ZO)-1 was determined by immunohistochemistry and immunoblotting.
  
  
  RESULTS
  Epithelial cells infected with RV showed 2-log more bound bacteria than sham-infected cultures, and bacteria were recovered from the basolateral media of RV- but not sham-infected cells. Infection of polarized airway epithelial cell cultures with RV for 24 hours caused a significant decrease in R(T) without causing cell death or apoptosis. Ultraviolet-treated RV did not decrease R(T), suggesting a requirement for viral replication. Reduced R(T) was associated with increased paracellular permeability, as determined by flux of fluorescein isothiocyanate (FITC)-inulin. Neutralizing antibodies to tumor necrosis factor (TNF)-alpha, IFN-gamma and IL-1beta reversed corresponding cytokine-induced reductions in R(T) but not that induced by RV, indicating that the RV effect is independent of these proinflammatory cytokines. Confocal microscopy and immunoblotting revealed the loss of ZO-1 from tight junction complexes in RV-infected cells. Intranasal inoculation of mice with RV1B also caused the loss of ZO-1 from the bronchial epithelium tight junctions in vivo.
  
  
  CONCLUSIONS
  RV facilitates binding, translocation, and persistence of bacteria by disrupting airway epithelial barrier function.
authors.affiliation: Department of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, MI 48109-0688, USA.
authors.forename: Umadevi
authors.initials: U
authors.lastname: Sajjan
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhao
authors.forename: Dieter C
authors.initials: DC
authors.lastname: Gruenert
authors.forename: Marc B
authors.initials: MB
authors.lastname: Hershenson
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001980:Bronchi / Q000378:metabolism / Q000473:pathology / Q000821:virology; D002463:Cell Membrane Permeability / Q000502:physiology*; D002478:Cells, Cultured; D015169:Colony Count, Microbial; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure / Q000821:virology; D019084:Fluorescent Antibody Technique, Indirect; D006192:Haemophilus Infections / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D006193:Haemophilus influenzae / Q000472:pathogenicity; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D010750:Phosphoproteins / Q000378:metabolism; D010850:Picornaviridae Infections / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D020545:Respiratory Mucosa / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D012229:Rhinovirus / Q000472:pathogenicity*; D029867:Xenopus Proteins; D062826:Zonula Occludens-1 Protein
chemical_list: D008565:Membrane Proteins; D010750:Phosphoproteins; C569045:TJP1 protein, Xenopus; C569042:TJP1 protein, human; C569043:Tjp1 protein, mouse; D029867:Xenopus Proteins; D062826:Zonula Occludens-1 Protein
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