Abstract：
BACKGROUND: Iguratimod (IGU) is widely used in clinical practice due to its stable anti-inflammatory effects. Our previous studies have confirmed that the proportion of Th17/Treg balance in patients taking IGU altered significantly. This study aims to explore the role of IGU in antibody-mediated rejection (ABMR) and its potential mechanisms.
METHODS: We conducted bioinformatics analysis of sequencing data from the GEO database to analyze the abundance of immune cell infiltration in transplanted kidney tissues. In vivo, IGU was intervened in a mice secondary skin transplantation model and a mice kidney transplantation ABMR model, and histological morphology of the grafts were examined by pathological staining, while relevant indicators were determined through qRT-PCR, immunohistochemistry, and enzyme-linked immunosorbent assay, observed T cell differentiation by flow cytometry, and preliminarily assessed the immunosuppressive effect of IGU. In vitro, we established Th17 and Treg cell induction and stimulation differentiation culture systems and added IGU for intervention to explore its effects on their differentiation.
RESULTS: Through bioinformatics analysis, we found that Th17 and Treg may play important roles in the occurrence and development of ABMR. In vivo, we found that IGU could effectively reduce the damage caused by ABMR to the grafts, alleviate the infiltration of inflammatory cells in the graft tissues, and reduce the deposition of C4d in the grafts. Moreover, it is also found that IGU regulated the differentiation of Th17 and Treg cells in the spleen and peripheral blood and reduced the expression of IL-17A in the grafts and serum. In addition, same changes were observed in the induction and differentiation culture system of Th17 and Treg cells in vitro after the addition of IGU.
CONCLUSIONS: IGU can inhibit the progression of ABMR by regulating the differentiation of Th17 and Treg cells, providing novel insights for optimizing clinical immunosuppressive treatment regimens.
METHODS: We conducted bioinformatics analysis of sequencing data from the GEO database to analyze the abundance of immune cell infiltration in transplanted kidney tissues. In vivo, IGU was intervened in a mice secondary skin transplantation model and a mice kidney transplantation ABMR model, and histological morphology of the grafts were examined by pathological staining, while relevant indicators were determined through qRT-PCR, immunohistochemistry, and enzyme-linked immunosorbent assay, observed T cell differentiation by flow cytometry, and preliminarily assessed the immunosuppressive effect of IGU. In vitro, we established Th17 and Treg cell induction and stimulation differentiation culture systems and added IGU for intervention to explore its effects on their differentiation.
RESULTS: Through bioinformatics analysis, we found that Th17 and Treg may play important roles in the occurrence and development of ABMR. In vivo, we found that IGU could effectively reduce the damage caused by ABMR to the grafts, alleviate the infiltration of inflammatory cells in the graft tissues, and reduce the deposition of C4d in the grafts. Moreover, it is also found that IGU regulated the differentiation of Th17 and Treg cells in the spleen and peripheral blood and reduced the expression of IL-17A in the grafts and serum. In addition, same changes were observed in the induction and differentiation culture system of Th17 and Treg cells in vitro after the addition of IGU.
CONCLUSIONS: IGU can inhibit the progression of ABMR by regulating the differentiation of Th17 and Treg cells, providing novel insights for optimizing clinical immunosuppressive treatment regimens.
摘要：
背景：Iguratimod（IGU）由于其稳定的抗炎作用而被广泛用于临床实践。我们先前的研究证实，服用IGU的患者中Th17/Treg平衡的比例发生了显着变化。本研究旨在探讨IGU在抗体介导的排斥反应(ABMR)中的作用及其潜在机制。
方法：我们对来自GEO数据库的测序数据进行了生物信息学分析，以分析移植肾组织中免疫细胞浸润的丰度。在体内,IGU干预小鼠二次皮肤移植模型和小鼠肾移植ABMR模型，通过病理染色检查移植物的组织学形态，同时通过qRT-PCR确定相关指标，免疫组织化学，和酶联免疫吸附测定，通过流式细胞术观察T细胞分化，并初步评价了IGU的免疫抑制作用。体外，我们建立了Th17和Treg细胞诱导和刺激分化培养系统，并加入IGU进行干预，以探讨其对其分化的影响。
结果：通过生物信息学分析，我们发现Th17和Treg可能在ABMR的发生发展中起重要作用。在体内,我们发现IGU可以有效地减少ABMR对移植物的损伤，减轻移植组织中炎性细胞的浸润，减少C4d在移植物中的沉积。此外，还发现IGU调节脾脏和外周血中Th17和Treg细胞的分化，并降低移植物和血清中IL-17A的表达。此外,添加IGU后，Th17和Treg细胞的体外诱导和分化培养系统观察到相同的变化。
结论：IGU可以通过调节Th17和Treg细胞的分化来抑制ABMR的进展，为优化临床免疫抑制治疗方案提供新的见解。
方法：我们对来自GEO数据库的测序数据进行了生物信息学分析，以分析移植肾组织中免疫细胞浸润的丰度。在体内,IGU干预小鼠二次皮肤移植模型和小鼠肾移植ABMR模型，通过病理染色检查移植物的组织学形态，同时通过qRT-PCR确定相关指标，免疫组织化学，和酶联免疫吸附测定，通过流式细胞术观察T细胞分化，并初步评价了IGU的免疫抑制作用。体外，我们建立了Th17和Treg细胞诱导和刺激分化培养系统，并加入IGU进行干预，以探讨其对其分化的影响。
结果：通过生物信息学分析，我们发现Th17和Treg可能在ABMR的发生发展中起重要作用。在体内,我们发现IGU可以有效地减少ABMR对移植物的损伤，减轻移植组织中炎性细胞的浸润，减少C4d在移植物中的沉积。此外，还发现IGU调节脾脏和外周血中Th17和Treg细胞的分化，并降低移植物和血清中IL-17A的表达。此外,添加IGU后，Th17和Treg细胞的体外诱导和分化培养系统观察到相同的变化。
结论：IGU可以通过调节Th17和Treg细胞的分化来抑制ABMR的进展，为优化临床免疫抑制治疗方案提供新的见解。
