title: P. aeruginosa drives CXCL8 synthesis via redundant toll-like receptors and NADPH oxidase in CFTR∆F508 airway epithelial cells.
abstract: BACKGROUND
  Understanding the mechanisms underlying bacterial-driven inflammation and neutrophil recruitment is important to design better therapies for CF. CXCL8 is an important chemokine found elevated in the airways of CF patients that recruits neutrophil to sites of the inflammation.
  
  
  METHODS
  Airway epithelial cells (AECs) expressing wild-type CFTR or CFTR∆F508 were challenged with Pseudomonas aeruginosa diffusible material (PsaDM) and the synthesis of CXCL8 was measured by quantitative real-time PCR and ELISA in absence or presence of MAPK inhibitors, TLR antagonists, glutathione and a NADPH oxidase inhibitor.
  
  
  RESULTS
  CFTR∆F508 AECs secrete more CXCL8 in response to PsaDM than their wild type counterpart, which can be reversed by addition of extracellular glutathione or incubating AECs at 27°C to favour folding and expression of CFTR at the cell membrane. Moreover, in CFTR∆F508 AECs, TLR2, TLR4 and TLR5 act redundantly to drive CXCL8 synthesis via the activation of NADPH oxidase.
  
  
  CONCLUSIONS
  These results demonstrate that NADPH oxidase is necessary for CXCL8 synthesis in response to TLRs activation by P. aeruginosa.
authors.affiliation: Meakins-Christie Laboratories, McGill University Heath Centre Research Institute, 3626 St-Urbain, Montréal, H2X 2P2, Canada.
authors.forename: Lucie
authors.initials: L
authors.lastname: Roussel
authors.forename: Guy
authors.initials: G
authors.lastname: Martel
authors.forename: Julie
authors.initials: J
authors.lastname: Bérubé
authors.forename: Simon
authors.initials: S
authors.lastname: Rousseau
mesh_terms: D002460:Cell Line; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000276:immunology; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism*; D020935:MAP Kinase Signaling System / Q000276:immunology; D019255:NADPH Oxidases / Q000378:metabolism*; D011552:Pseudomonas Infections / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology / Q000201:enzymology / Q000382:microbiology; D051195:Toll-Like Receptor 2 / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000378:metabolism; D051198:Toll-Like Receptor 5 / Q000378:metabolism; D051193:Toll-Like Receptors / Q000378:metabolism
chemical_list: C505032:CFTR protein, human; C508606:CXCL8 protein, human; D016209:Interleukin-8; D017382:Reactive Oxygen Species; C495342:TLR2 protein, human; C495345:TLR4 protein, human; C495346:TLR5 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051198:Toll-Like Receptor 5; D051193:Toll-Like Receptors; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019255:NADPH Oxidases