Institute of Medical Immunology, Affiliated Hospital of Jiangsu University, Zhenjiang, 212000, China.
Department of Laboratory Medicine, Jiangsu Key Laboratory of Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang, China.
Inflamm Res. 2022 Dec;71(12):1577-1588. doi: 10.1007/s00011-022-01660-5. Epub 2022 Oct 27.
Recent studies have revealed a role of the ligand for glucocorticoid-induced TNFR family-related protein (GITRL) in mediating functional dysregulations of myeloid-derived suppressor cells (MDSCs) in the pathogenesis of primary Sjögren syndrome (pSS), but the underlying molecular mechanism is largely unclear. In this study, we aimed to elucidate GITRL-mediated signaling pathways in MDSCs during the development of experimental SS (ESS).
MDSCs were stimulated with recombinant GITRL, the activation of PTEN, AKT and STAT3 in MDSCs was analyzed by Western blot. MDSCs with different treatment were adoptively transferred to ESS mice. ELISA was used to detect the level of autoantibodies. Proportions of Th1 and Th17 cells were examined by flow cytometry. Histological evaluation of glandular destruction was analyzed by hematoxylin and eosin (HE) staining. The interaction of GITR, TRAF3 and PP2A was detected by CoIP.
Upon the engagement of GITR on MDSCs, PTEN was activated and led to the inhibition of downstream AKT/STAT3 signaling pathway, therefore, resulting in the impaired immunosuppressive function of MDSCs. In ESS mice, blocking the activity of PTEN could efficiently restore the immunomodulatory effect of MDSCs and alleviate the progression of ESS. Furthermore, TRAF3 was found to bind to GITR, and then recruited PP2A to dephosphorylate PTEN, thus enhancing the activity of PTEN.
This study elucidated the molecular mechanism underlying the effect of GITRL in regulating the function of MDSCs, which may provide a new therapeutic target for the treatment of pSS.
最近的研究表明,糖皮质激素诱导的肿瘤坏死因子受体家族相关蛋白(GITRL)配体在介导原发性干燥综合征(pSS)发病机制中的髓系来源抑制细胞(MDSCs)功能失调中起作用,但潜在的分子机制尚不清楚。在这项研究中,我们旨在阐明实验性干燥综合征(ESS)发展过程中 GITRL 介导的 MDSC 信号通路。
用重组 GITRL 刺激 MDSCs,通过 Western blot 分析 MDSCs 中 PTEN、AKT 和 STAT3 的激活情况。用不同处理方法的 MDSCs 过继转移到 ESS 小鼠中。用 ELISA 检测自身抗体水平。用流式细胞术检测 Th1 和 Th17 细胞的比例。用苏木精和伊红(HE)染色分析腺体破坏的组织学评价。通过 CoIP 检测 GITR、TRAF3 和 PP2A 的相互作用。
GITR 在 MDSCs 上结合后,PTEN 被激活,导致下游 AKT/STAT3 信号通路受到抑制,从而导致 MDSCs 的免疫抑制功能受损。在 ESS 小鼠中,阻断 PTEN 的活性可有效恢复 MDSCs 的免疫调节作用并减轻 ESS 的进展。此外,发现 TRAF3 与 GITR 结合,然后募集 PP2A 使 PTEN 去磷酸化,从而增强 PTEN 的活性。
本研究阐明了 GITRL 调节 MDSCs 功能的作用的分子机制,这可能为治疗 pSS 提供新的治疗靶点。