Wu Shiyao, Xie Yanli, Jiang Ying, Zhang Xiaoli, Zhou Yaou, Zuo Xiaoxia, Li Tong
Department of Rheumatology and Immunology, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Department of Dermatology and Immunology, National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Clin Rheumatol. 2025 Mar;44(3):989-998. doi: 10.1007/s10067-025-07320-3. Epub 2025 Jan 15.
Previous research has demonstrated ɑ7nAch receptor (ɑ7nAchR) agonists to provide benefit for rheumatoid arthritis (RA) patients. However, the immunological mechanism of action for these ɑ7nAchR agonists has not been elucidated. Herein, the effect of GTS-21, a selective ɑ7nAchR agonist, on the differentiation of Th17 and Th2 cells was assessed. CD4 + T cells were obtained from the peripheral blood mononuclear cells (PBMCs) of RA patients and healthy donors. CD4 + T cells were separately differentiated into Th2 or Th17 cells with or without GTS-21 and with or without alpha-bungarotoxin (ɑBgt) (a ɑ7nAchR antagonist). The proportions of Th17 and Th2 cells were assessed by flow cytometry. Levels of the T cell cytokines, IL-17A and IL-4, were assessed by ELISA. Specific transcription factors, retinoic orphan receptor c (RORc), and GATA Binding Protein 3 (GATA-3) were detected by western blot. GTS-21 reduced IL-17A and increased IL-4 production by RA PBMCs. GTS-21 reduced the percentage of Th17 cells and increased the percentage of Th2 cells during Th17 and Th2 differentiation, respectively. GTS-21 downregulated RA CD4 + T cells RORc levels and reduced the secretion of IL-17A during Th17 differentiation. GTS-21 upregulated RA CD4 + T cells GATA3 and promoted IL-4 production during Th2 differentiation. ɑ-Bgt blocked the effects of GTS-21 during Th17 and Th2 differentiation. These results demonstrated that GTS-21 suppressed RA Th17 differentiation and promoted Th2 differentiation. As such, the use of GTS-21 may be a new therapeutic approach by which to treat RA patients. Key Points • GTS-21 suppressed RA Th17 differentiation and promoted Th2 differentiation via acting on ɑ7nAchR. • The protective effect of GTS-21 on RA may be related to its regulation of Th cell subsets.
先前的研究已证明α7烟碱型乙酰胆碱受体（α7nAchR）激动剂对类风湿关节炎（RA）患者有益。然而，这些α7nAchR激动剂的免疫作用机制尚未阐明。在此，评估了选择性α7nAchR激动剂GTS-21对Th17和Th2细胞分化的影响。从RA患者和健康供体的外周血单个核细胞（PBMC）中获取CD4 + T细胞。在有或没有GTS-21以及有或没有α-银环蛇毒素（αBgt，一种α7nAchR拮抗剂）的情况下，将CD4 + T细胞分别分化为Th2或Th17细胞。通过流式细胞术评估Th17和Th2细胞的比例。通过酶联免疫吸附测定（ELISA）评估T细胞细胞因子IL-17A和IL-4的水平。通过蛋白质免疫印迹法检测特异性转录因子维甲酸孤儿受体c（RORc）和GATA结合蛋白3（GATA-3）。GTS-21降低了RA PBMC产生的IL-17A并增加了IL-4的产生。在Th17和Th2分化过程中，GTS-21分别降低了Th17细胞的百分比并增加了Th2细胞的百分比。GTS-21下调了RA CD4 + T细胞的RORc水平，并在Th17分化过程中减少了IL-17A的分泌。GTS-21上调了RA CD4 + T细胞的GATA3，并在Th2分化过程中促进了IL-4的产生。α-Bgt在Th17和Th2分化过程中阻断了GTS-21的作用。这些结果表明，GTS-21抑制RA Th17分化并促进Th2分化。因此，使用GTS-21可能是治疗RA患者的一种新的治疗方法。要点•GTS-21通过作用于α7nAchR抑制RA Th17分化并促进Th2分化。•GTS-21对RA的保护作用可能与其对Th细胞亚群的调节有关。
