Polarization of human gingival fibroblasts by Th1‐, Th2‐, Th17‐, and Treg‐derived cytokines

Abstract

Background and objective The purpose of this study was to evaluate whether gingival fibroblasts (GFs) can be differently activated and polarized into distinct functional subtypes by T‐helper (Th) cytokines. Methods Gingival fibroblasts were stimulated with interferon (IFN)‐γ, interleukin (IL)‐4, IL‐17, and transforming growth factor (TGF)‐β, representative cytokines of Th1, Th2, Th17, and regulatory T cells, respectively, and the gene expression profiles were analyzed by microarray. Differentially expressed genes (DEGs) in GFs stimulated by 4 cytokines were screened, and a gene ontology (GO) analysis of the DEGs was conducted. To confirm the reliability of the microarray results, the DEGs that showed the largest differences compared with non‐stimulated GFs were further analyzed by RT‐PCR. To evaluate the effect of polarization on GFs responses to lipopolysaccharide (LPS), GFs stimulated by 4 cytokines were further stimulated with Escherichia coli LPS and mRNA levels of several genes were analyzed using RT‐PCR. Results Differentially expressed genes by 4 Th cytokines were enriched in different GO terms, and the patterns of gene expression on GFs were shown functionally different. GFs stimulated with IFN‐γ (GF(IFN‐γ)) up‐regulated the expression of chemokines (chemokine (C‐X‐C motif) ligand (CXCL)9, −10, −11, chemokine (C‐C motif) ligand (CCL)8), molecules involved in antigen presentation, complement component 3 (C3), and other immune response–related molecules, whereas they down‐regulated the expression of several types of collagen, extracellular matrix (ECM) components, and DNA replication and nuclear protein–related molecules. By contrast, GF(IL‐4) up‐regulated the expression of ECM components, cell adhesion molecules, and tissue development–related molecules and down‐regulated the expression of chemokines (CXCL10 and CXCL8) and adaptive immune response–related molecules. GF(IL‐17) up‐regulated the expression of chemokines and other molecules for neutrophil infiltration and activation, the pro‐inflammatory cytokine IL‐6, and C3. GF(TGF‐β) up‐regulated the expression of cell growth–related molecules, ECM components, several types of collagen, and cell adhesion molecules and down‐regulated the expression of molecules related to complement activation and bacterial recognition. GFs stimulated by 4 cytokines responded differently to LPS. Conclusion These results show that Th cytokines can polarize GFs into cells with functionally distinct features: immune‐activating but tissue‐destructive GF(IFN‐γ), tissue‐reparative, and immune‐inhibiting GF(IL‐4), highly pro‐inflammatory GF(IL‐17), and potent tissue‐reparative GF(TGF‐β).

The Role of Gingival Fibroblasts in the Pathogenesis of Periodontitis

“…Furthermore, the involvement of nuclear factor–κB (NF-κB) and mitogen-activated protein (MAP) kinases in this process has been thoroughly characterized (Båge et al 2010). Interestingly, the cytokines interferon-γ (IFN-γ) and IL-4 skew polarized GFs into functionally distinct subtypes that differ in their potential to activate immune cells and effect tissue repair (Ha et al 2022), as well as their responses to oral pathogens (Jang et al 2021). Although these in vitro phenotypes represent the extremes of the polarization spectrum and are unlikely to exist in vivo, single-cell analysis of the healthy oral mucosa identified transcriptionally distinct subtypes of oral fibroblasts that are characterized by gene signatures associated either with inflammatory responses or with collagen synthesis and matrix remodeling (Williams et al 2021).…”

An allergenic plant calmodulin from Artemisia pollen primes human DCs leads to Th2 polarization

“…DCs play a crucial role in the priming and differentiation of CD4+ T cells into several distinct subsets, including Th1, Th2, and Th17, as well as regulatory T cells (Treg) ( 33 ). Categorization of Th cells is usually based on a dominant cytokine or even a family of cytokines, including Th1 (IL-6, INF-γ, TNF-α), Th2 (IL-4, IL-5, IL-6, IL-10, IL-13), Th17 (IL-17, IL-21, IL-22), and Treg (TGF-β, IL-10, IL-35) ( 34 ) ( Figure 4A ). Here, we monitored the dominant cytokine levels to determine the Th polarization.…”

CD81+fibroblasts, a unique subpopulation with accelerated cellular senescence, exaggerate inflammation and activate neutrophils via C3/C3aR1 axis in periodontitis

“…Genetic analysis and preclinical studies have confirmed C3 as a potential pharmacological target for periodontitis treatment (Alayash et al, 2023; Hajishengallis et al, 2021). gingival fibroblasts stimulated with IFN-γ (GF(IFN-γ)) up-regulated the expression of chemokines (CXCL9, -10, -11, CCL8), molecules involved in antigen presentation, complement component 3 (C3), and other immune response-related molecules (Ha et al, 2022). Our experimental results have demonstrated that CD81+ gingival fibroblasts are an important source of C3.…”