title: Hypothiocyanite produced by human and rat respiratory epithelial cells inactivates extracellular H1N2 influenza A virus.
abstract: OBJECTIVE
  Our aim was to study whether an extracellular, oxidative antimicrobial mechanism inherent to tracheal epithelial cells is capable of inactivating influenza H1N2 virus.
  
  
  METHODS
  Epithelial cells were isolated from tracheas of male Sprague-Dawley rats. Both primary human and rat tracheobronchial epithelial cells were differentiated in air-liquid interface cultures.
  
  
  METHODS
  A/swine/Illinois/02860/09 (swH1N2) influenza A virions were added to the apical side of airway cells for 1 h in the presence or absence of lactoperoxidase or thiocyanate.
  
  
  METHODS
  Characterization of rat epithelial cells (morphology, Duox expression) occurred via western blotting, PCR, hydrogen peroxide production measurement and histology. The number of viable virions was determined by plaque assays. Statistical difference of the results was analyzed by ANOVA and Tukey's test.
  
  
  RESULTS
  Our data show that rat tracheobronchial epithelial cells develop a differentiated, polarized monolayer with high transepithelial electrical resistance, mucin production and expression of dual oxidases. Influenza A virions are inactivated by human and rat epithelial cells via a dual oxidase-, lactoperoxidase- and thiocyanate-dependent mechanism.
  
  
  CONCLUSIONS
  Differentiated air-liquid interface cultures of rat tracheal epithelial cells provide a novel model to study airway epithelium-influenza interactions. The dual oxidase/lactoperoxidase/thiocyanate extracellular oxidative system producing hypothiocyanite is a fast and potent anti-influenza mechanism inactivating H1N2 viruses prior to infection of the epithelium.
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Gingerich
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Lan
authors.initials: L
authors.lastname: Pang
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Jarod
authors.initials: J
authors.lastname: Hanson
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Dlugolenski
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Streich
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Eric R
authors.initials: ER
authors.lastname: Lafontaine
authors.affiliation: Department of Pathology, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Tamás
authors.initials: T
authors.lastname: Nagy
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA.
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Tripp
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, 501 D.W. Brooks Drive, Athens, GA, 30602, USA. radab@uga.edu.
authors.forename: Balázs
authors.initials: B
authors.lastname: Rada
mesh_terms: D000818:Animals; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D057221:Influenza A Virus, H1N2 Subtype / Q000187:drug effects*; D007784:Lactoperoxidase / Q000378:metabolism; D061985:Madin Darby Canine Kidney Cells; D008297:Male; D009077:Mucins / Q000096:biosynthesis; D061251:Primary Cell Culture; D051381:Rats; D017207:Rats, Sprague-Dawley; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism*; D013861:Thiocyanates / Q000378:metabolism*
chemical_list: D009077:Mucins; D013861:Thiocyanates; C014607:hypothiocyanite ion; D006861:Hydrogen Peroxide; D007784:Lactoperoxidase
keywords: Airway epithelium; Dual oxidase; Hydrogen peroxide; Hypothiocyanite; Influenza