IFN-γ or IL-4 polarization impacts the response of gingival fibroblasts to oral bacteria

Abstract

Background and objective: We previously reported that gingival fibroblasts (GFs) can be polarized into functionally distinct subtypes, immune-activating but tissue-destructive or tissue-reparative, in response to T helper (Th1) and Th2 stimuli, respectively. The purpose of this study was to evaluate the effect of polarization on GFs responses to oral bacteria. Methods: Unprimed (GF(−)) and IFN-γ (GF(IFN-γ)) or IL-4 primed (GF(IL-4)) GFs were stimulated with live Fusobacterium nucleatum or Porphyromonas gingivalis. The mRNA expression of IL-1β, IL-4, LPS-recognizing components (Toll-like receptor (TLR) 4, CD14), molecules involved in antigen presentation (human leukocyte antigen (HLA)-ABC, HLA-DP, CD74, CD40), chemokines (C-X-C motif chemokine (CXCL)10, CXCL11, chemokine (C-C motif) ligand 20 (CCL20)), collagen type 1 alpha 1 (COL1A1), and matrix metalloproteinase (MMP)-1, and the protein levels of IL-1β, CD14, CXCL11, CCL20, and COL1A1 accumulated in supernatants were analyzed using real-time PCR and ELISA. Results: In response to oral bacteria, the GF(IFN-γ) significantly upregulated the expression of LPS-recognizing components, molecules involved in antigen presentation, CXCL10, and CXCL11, whereas the levels of IL-4 and COL1A1 were downregulated, compared with GF(−). The levels of IL-1β, CCL20, and MMP-1 from GF(IFN-γ) were differently regulated between both bacteria; F. nucleatum was synergistically upregulated, but P. gingivalis was downregulated. The GF(IL-4) stimulated with both bacteria upregulated the levels of IL-4, whereas the levels of TLR4 and chemokines were downregulated, compared with GF(−). The regulation of IL-1β, CD14, CXCL11, CCL20, and COL1A1 proteins showed a similar tendency with mRNA regulation. Conclusion: Polarization of GFs with IFN-γ or IL-4 affected the way that GFs responded to oral bacteria through up or downregulation of inflammatory responses and extracellular matrix control.