Interferon-γ stimulates CD14, TLR2 and TLR4 mRNA expression in gingival fibroblasts increasing responsiveness to bacterial challenge. (January 2016)

• Record Type: Journal Article
• Authors: Lappin, M.J.; Brown, V.; Zaric, S.S.; Lundy, F.T.; Coulter, W.A.; Irwin, C.R.
• Is Part Of: Archives of oral biology. Volume 61(2016)
• Page Start: 36
• Page End: 43
• Publication Date: 2016-01
• Subjects: IFN-γ -- CD14 -- Fibroblast -- Lipopolysaccharide; Mouth -- Periodicals; Mouth -- Diseases -- Periodicals; Dentistry -- Periodicals; Electronic journals; 617.6005
• DOI: 10.1016/j.archoralbio.2015.10.005
• Languages: English
• ISSNs: 0003-9969

Abstract

Highlights: IFN-γ induces mRNA expression of CD14, TLR2 and TLR4 in human gingival fibroblasts. IFN-γ induces expression of membrane-bound CD14 in human gingival fibroblasts. Pre-incubation with IFN-γ followed by LPS stimulation increases cytokine production. LPS-induced cytokine production is abrogated by a blocking antibody to CD14. IFN-γ primes human gingival fibroblasts, through up-regulated CD14 expression. Abstract: Objective: To investigate the potential effects of IFN-03A5 on the responsiveness of human gingival fibroblasts to bacterial challenge. Design: 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. Conclusion: 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.