The regulation of the Treg/Th17 balance by mesenchymal stem cells in human systemic lupus erythematosus
The regulation of the Treg/Th17 balance by mesenchymal stem cells in human systemic lupus erythematosus
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间充质干细胞对人系统性红斑狼疮Treg/Th17平衡的调节
DOI：
10.1038/cmi.2015.89
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发表时间：
2017-05-01
影响因子：
24.1
通讯作者：
Sun, Lingyun
中科院分区：
文献类型：
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作者：
Wang, Dandan;Huang, Saisai;Sun, Lingyun
背景和目的脐带（UC）来源的间充质干细胞（MSC）已显示出多种免疫细胞的免疫调节作用。本研究的目的是探讨UC MSCs调节系统性红斑狼疮(SLE)患者外周调节性T细胞(Treg)和辅助性T细胞17(Th17)细胞的机制。方法对30例常规治疗无效的活动性SLE患者进行UC MSCs输注。 MSCT移植后1周、1个月、3个月、6个月、12个月时测定外周血CD4+CD25+Foxp3+调节性T细胞(Treg)和CD3+CD8-IL17A+Th17细胞的百分比以及Foxp3和IL-17的平均荧光强度(MFI)。使用ELISA检测血清细胞因子，包括转化生长因子β（TGF-β）、肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）和IL-17A。收集患者外周血单个核细胞,分别按1:1、10:1、50:1的比例与UC MSC共培养72 h,检测Treg、Th17细胞比例,流式细胞术测定Foxp3、IL-17的MFI。采用ELISA法检测上清液中的细胞因子。在共培养体系中添加针对TGF-β、IL-6、吲哚胺2、3-双加氧酶(IDO)、前列腺素E2的抑制剂，观察Treg和Th17细胞的百分比。结果UC MSC移植后1周、1个月、3个月外周血Treg和Foxp3 MFI百分比增加，而Th17比例和IL-17 MFI下降治疗后3个月、6个月和12个月，血清TGF-β在治疗1周、3个月和12个月时增加，血清TNF-α从1周开始减少。 MSCT前后血清IL-6和IL-17A没有变化。体外研究表明，UC MSCs 剂量依赖性上调 SLE 患者外周 Treg 比例，且不依赖于细胞间接触。然而，Th17细胞的下调不具有剂量依赖性，也不依赖于细胞与细胞的接触。共培养后上清液TGF-β和IL-6水平显着升高，TNF-α显着降低，但IL-17A无变化。添加抗TGF-β抗体显着消除了Treg细胞的上调，添加PGE2抑制剂显着消除了Th17细胞的下调。抗IL-6抗体和IDO抑制剂均对Treg和Th17细胞没有影响。结论UC MSCs通过调节狼疮患者的TGF-β和PGE2上调Treg细胞并下调Th17细胞。
Background and objectiveUmbilical cord (UC)-derived mesenchymal stem cells (MSCs) have shown immunoregulation of various immune cells. The aim of this study was to investigate the mechanism of UC MSCs in the regulation of peripheral regulatory T cells (Treg) and T helper 17 (Th17) cells in patients with systemic lupus erythematosus (SLE).MethodsThirty patients with active SLE, refractory to conventional therapies, were given UC MSCs infusions. The percentages of peripheral blood CD4+ CD25+ Foxp3+ regulatory T cells (Treg) and CD3+ CD8-IL17A+ Th17 cells and the mean fluorescence intensities (MFI) of Foxp3 and IL-17 were measured at 1 week, 1 month, 3 months, 6 months, and 12 months after MSCs transplantation (MSCT). Serum cytokines, including transforming growth factor beta (TGF-β), tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and IL-17A were detected using ELISA. Peripheral blood mononuclear cells from patients were collected and co-cultured with UC MSCs at ratios of 1: 1, 10: 1, and 50: 1, respectively, for 72 h to detect the proportions of Treg and Th17 cells and the MFIs of Foxp3 and IL-17 were determined by flow cytometry. The cytokines in the supernatant solution were detected using ELISA. Inhibitors targeting TGF-β, IL-6, indoleamine 2, 3-dioxygenase (IDO), and prostaglandin E2 were added to the co-culture system, and the percentages of Treg and Th17 cells were observed.ResultsThe percentage of peripheral Treg and Foxp3 MFI increased 1 week, 1 month, and 3 months after UC MSCs transplantation, while the Th17 proportion and MFI of IL-17 decreased 3 months, 6 months, and 12 months after the treatment, along with an increase in serum TGF-β at 1 week, 3 months, and 12 months and a decrease in serum TNF-α beginning at 1 week. There were no alterations in serums IL-6 and IL-17A before or after MSCT. In vitro studies showed that the UC MSCs dose-dependently up-regulated peripheral Treg proportion in SLE patients, which was not depended on cell–cell contact. However, the down-regulation of Th17 cells was not dose-dependently and also not depended on cell–cell contact. Supernatant TGF-β and IL-6 levels significantly increased, TNF-α significantly decreased, but IL-17A had no change after the co-culture. The addition of anti-TGF-β antibody significantly abrogated the up-regulation of Treg, and the addition of PGE2 inhibitor significantly abrogated the down-regulation of Th17 cells. Both anti-IL-6 antibody and IDO inhibitor had no effects on Treg and Th17 cells.ConclusionsUC MSCs up-regulate Treg and down-regulate Th17 cells through the regulation of TGF-β and PGE2 in lupus patients.