Department of Periodontology, Institute of Oral Health Science, Ajou University School of Medicine, Suwon, Korea.
J Periodontal Res. 2022 Jun;57(3):487-501. doi: 10.1111/jre.12978. Epub 2022 Feb 25.
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.
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.
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.
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).
本研究旨在评估辅助性 T 细胞（Th）细胞因子是否可以使牙龈成纤维细胞（GFs）分别被激活和极化成为不同的功能亚型。
用干扰素（IFN）-γ、白细胞介素（IL）-4、IL-17 和转化生长因子（TGF）-β分别刺激 GFs，这 4 种细胞因子分别代表 Th1、Th2、Th17 和调节性 T 细胞。通过微阵列分析细胞因子刺激后 GFs 的基因表达谱。筛选出 4 种细胞因子刺激的 GFs 中差异表达的基因（DEGs），并对 DEGs 进行基因本体（GO）分析。为了确认微阵列结果的可靠性，进一步通过 RT-PCR 分析与非刺激 GFs 相比差异最大的 DEGs。为了评估极化对 GFs 对脂多糖（LPS）反应的影响，进一步用 4 种细胞因子刺激 GFs，然后用大肠杆菌 LPS 刺激，并用 RT-PCR 分析几种基因的 mRNA 水平。
4 种 Th 细胞因子诱导的差异表达基因在不同的 GO 术语中富集，GFs 的基因表达模式表现出不同的功能。IFN-γ 刺激的 GFs（GF（IFN-γ））上调趋化因子（趋化因子（C-X-C 基序）配体（CXCL）9、-10、-11、趋化因子（C-C 基序）配体（CCL）8）、抗原呈递分子、补体成分 3（C3）和其他免疫反应相关分子的表达，而下调几种类型的胶原、细胞外基质（ECM）成分和与 DNA 复制和核蛋白相关的分子的表达。相比之下，IL-4 刺激的 GFs（GF（IL-4））上调 ECM 成分、细胞粘附分子和组织发育相关分子的表达，下调趋化因子（CXCL10 和 CXCL8）和适应性免疫反应相关分子的表达。IL-17 刺激的 GFs 上调趋化因子和其他用于中性粒细胞浸润和激活的分子、促炎细胞因子 IL-6 和 C3 的表达。TGF-β 刺激的 GFs 上调与细胞生长相关的分子、ECM 成分、几种类型的胶原和细胞粘附分子的表达，下调补体激活和细菌识别相关分子的表达。4 种细胞因子刺激的 GFs 对 LPS 的反应不同。
这些结果表明，Th 细胞因子可以将 GFs 极化成为具有不同功能特征的细胞：具有免疫激活但组织破坏性的 GF（IFN-γ）、组织修复和免疫抑制的 GF（IL-4）、高度促炎的 GF（IL-17）和有效的组织修复的 GF（TGF-β）。
