title: Pseudomonas aeruginosa Inhibition of Flagellin-activated NF-kappaB and interleukin-8 by human airway epithelial cells.
abstract: Pseudomonas aeruginosa-induced activation of NF-kappaB and secretion of proinflammatory cytokines by airway epithelial cells require that the bacteria express flagellin. We tested whether P. aeruginosa and human airway epithelial cells secrete factors that modulated this response. Experiments were performed with both the Calu-3 cell line and primary cultures of tracheal epithelial cells. P. aeruginosa strain PAK DeltafliC (flagellin knockout) did not activate NF-kappaB or interleukin-8 (IL-8) but inhibited flagellin-activated NF-kappaB by 40 to 50% and IL-8 secretion by 20 to 25%. PAK DeltafliC also inhibited NF-kappaB induced by IL-1beta and Toll-like receptor 2 agonist Pam3CSK4. Similar inhibitions were observed with strains PAK, PAO1, and PA14. The inhibitory factor was present in conditioned medium isolated from PAK DeltafliC or Calu-3 plus PAK DeltafliC, but it was not present in conditioned medium isolated from Calu-3 cells alone or from PAK DeltafliC that had been heat treated. Inhibition by PAK DeltafliC-conditioned medium was exerted from either the apical or the basolateral side of the epithelium, was enhanced in simple Ringer's solution over that in tissue culture medium, and did not result from altered pH or depletion of glucose. The inhibitory effect of conditioned medium was abolished by boiling and appeared from filtration studies to result from effects of a factor with a molecular mass of <3 kDa. These and further studies with isogenic mutants led to the conclusion that the NF-kappaB and IL-8 response of airway epithelial cells to P. aeruginosa results from a balance of proinflammatory effects of flagellin and antiinflammatory effects of a small (<3-kDa), heat-sensitive factor(s) that is not lipopolysaccharide, C12 homoserine lactone, alginate, CIF, or exotoxin A, S, T, U, or Y.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3200, USA.
authors.forename: Jose
authors.initials: J
authors.lastname: Pena
authors.forename: Zhu
authors.initials: Z
authors.lastname: Fu
authors.forename: Christian
authors.initials: C
authors.lastname: Schwarzer
authors.forename: Terry E
authors.initials: TE
authors.lastname: Machen
mesh_terms: D002460:Cell Line; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry; D004847:Epithelial Cells / Q000276:immunology* / Q000382:microbiology*; D005408:Flagellin / Q000235:genetics / Q000276:immunology*; D017353:Gene Deletion; D006801:Humans; D053583:Interleukin-1beta / Q000037:antagonists & inhibitors; D016209:Interleukin-8 / Q000037:antagonists & inhibitors*; D055666:Lipopeptides / Q000037:antagonists & inhibitors; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D020545:Respiratory Mucosa
chemical_list: C508606:CXCL8 protein, human; D017077:Culture Media, Conditioned; D053583:Interleukin-1beta; D016209:Interleukin-8; D055666:Lipopeptides; D016328:NF-kappa B; C508075:Pam(3)CSK(4) peptide; D005408:Flagellin