title: Interferon-γ stimulates CD14, TLR2 and TLR4 mRNA expression in gingival fibroblasts increasing responsiveness to bacterial challenge.
abstract: OBJECTIVE
  To investigate the potential effects of IFN-03A5 on the responsiveness of human gingival fibroblasts to bacterial challenge.
  
  
  METHODS
  mRNA and protein expression of CD14, TLR2 and TLR4 in human gingival fibroblasts was detected by quantitative polymerase chain reaction (Q-PCR) and flow cytometry. The effect of preincubation with IFN-03A5 on subsequent bacterial LPS-induced expression of IL-6 and IL-8 by gingival fibroblasts was determined by ELISA. Bacterial LPS-induced IκBα degradation in human gingival fibroblasts was investigated by western blot.
  
  
  RESULTS
  Human gingival fibroblasts express CD14, TLR2 and TLR4 mRNAs. IFN-03A5, but not IL-103B2, induced mRNA expression of all three receptors and the expression of membrane bound CD14 protein. Pre-incubation of fibroblasts with IFN-03A5 and subsequent stimulation with Escherichia coli LPS or Porphyromonas gingivalis LPS led to increased production of IL-6 and IL-8. LPS-induced pro-inflammatory cytokine production was abrogated by a blocking antibody to CD14. Both E. coli LPS and P. gingivalis LPS induced IκBα degradation in human gingival fibroblasts.
  
  
  CONCLUSIONS
  Our data indicate that IFN-03A5 primes human gingival fibroblasts, through the upregulation of CD14 expression, which results in increased responsiveness to bacterial LPS challenge, as determined by pro-inflammatory cytokine production.
authors.affiliation: Centre for Dentistry, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, UK. Electronic address: m.lappin@qub.ac.uk.
authors.forename: M J
authors.initials: MJ
authors.lastname: Lappin
authors.affiliation: Centre for Infection and Immunity, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, UK.
authors.forename: V
authors.initials: V
authors.lastname: Brown
authors.affiliation: Centre for Infection and Immunity, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, UK.
authors.forename: S S
authors.initials: SS
authors.lastname: Zaric
authors.affiliation: Centre for Infection and Immunity, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, UK.
authors.forename: F T
authors.initials: FT
authors.lastname: Lundy
authors.affiliation: Centre for Dentistry, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, UK.
authors.forename: W A
authors.initials: WA
authors.lastname: Coulter
authors.affiliation: Centre for Dentistry, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, UK.
authors.forename: C R
authors.initials: CR
authors.lastname: Irwin
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000276:immunology; D005347:Fibroblasts / Q000187:drug effects*; D005434:Flow Cytometry; D005881:Gingiva / Q000166:cytology*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D018950:Lipopolysaccharide Receptors / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D051195:Toll-Like Receptor 2 / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D007371:Interferon-gamma
keywords: CD14; Fibroblast; IFN-γ; Lipopolysaccharide