Autophagy enhances mesenchymal stem cell-mediated CD4+ T cell migration and differentiation through CXCL8 and TGF-β1
Autophagy enhances mesenchymal stem cell-mediated CD4+ T cell migration and differentiation through CXCL8 and TGF-β1
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DOI：
10.1186/s13287-019-1380-0
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发表时间：
2019-08-23
影响因子：
7.5
通讯作者：
Wu, Yanfeng
中科院分区：
文献类型：
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作者：
Cen, Shuizhong;Wang, Peng;Wu, Yanfeng
背景：间充质干细胞(MSCs)已被认为是治疗各种炎症性疾病和自身免疫性疾病的一种有前途的工具。应激状态影响免疫介导的治疗，并激活MSCs的自噬。然而，自噬是否影响MSC介导的CD4(+)T细胞的募集和分化仍不清楚。方法用3-甲基腺嘌呤(3-MA)和雷帕霉素处理骨髓间充质干细胞以调节自噬，然后与CD4(+)T细胞共培养。流式细胞仪检测CD4(+)T细胞的迁移和分化。此外，利用实时定量聚合酶链式反应分析了已知趋化因子的基因表达水平。采用双抗体夹心法和免疫印迹法检测C-X-C基序、趋化因子配体8(CXCL8)和转化生长因子-β1蛋白水平。外源抗体和短发夹状RNA被用来调节CXCL8和转化生长因子-β1的水平，这使我们能够评估自噬如何影响MSC介导的CD4(+)T细胞的迁移和分化。结果3-MA抑制骨髓间充质干细胞的自噬，雷帕霉素可激活自噬。雷帕霉素促进CD4(+)T细胞的迁移，而3-MA则抑制其迁移。从机制上讲，我们发现自噬增强了CXCL8的分泌，外源性CXCL8和抗CXCL8抗体的加入消除了组间CD4(+)T细胞迁移的差异。此外，与对照组相比，经雷帕霉素处理的MSCs的调节性T细胞(Treg)比例增加，而辅助性T细胞(Th1)比例降低，而经3-MA处理的MSCs的作用与对照组相反。转化生长因子-β1的过度表达和慢病毒敲除纠正了不同组之间Treg和Th1细胞比率的差异。结论间充质干细胞自噬分别通过CXCL8和转化生长因子-β1介导了CD4(+)T细胞的迁移和分化。这些结果为改进MSC介导的治疗提供了一种潜在的新策略。
Background Mesenchymal stem cells (MSCs) have been recognized as a promising tool for the treatment of various inflammatory disorders and autoimmune diseases. Stress conditions affect immune-mediated treatment and activate autophagy in MSCs. However, whether autophagy affects the MSC-mediated recruitment and differentiation of CD4(+) T cells remains elusive. Methods MSCs were pretreated with 3-methyladenine (3-MA) and rapamycin to regulate autophagy, and then co-cultured with CD4(+) T cells. CD4(+) T cell migration and differentiation were detected by flow cytometry. Further, gene expression levels of well-known chemokines were analyzed by quantitative real-time PCR. Enzyme-linked immunosorbent assays and western blot analysis were performed to detect C-X-C motif chemokine ligand 8 (CXCL8) and transforming growth factor (TGF)-beta 1 protein levels. An exogenous antibody and short hairpin RNA were used to regulate CXCL8 and TGF-beta 1 levels, which enabled us to evaluate how autophagy affected MSC-mediated CD4(+) T cell migration and differentiation. Results 3-MA inhibited autophagy in MSCs, which was activated by rapamycin. Rapamycin increased the migration of CD4(+) T cells, whereas 3-MA decreased their migration. Mechanistically, we found that autophagy strengthened CXCL8 secretion, and the addition of exogenous CXCL8 and an anti-CXCL8 antibody eliminated the difference of CD4(+) T cell migration among groups. Further, the ratio of regulatory T (Treg) cells was increased in rapamycin-pretreated MSCs, but the ratio of T helper 1 (Th1) cells was decreased, while pretreatment of MSCs with 3-MA induced the opposite effect compared with the control group. TGF-beta 1 overexpression and knockdown using lentiviruses rectified the differences in the ratios of Treg and Th1 cells among the groups. Conclusion This study demonstrates that autophagy of mesenchymal stem cells mediates CD4(+) T cell migration and differentiation through CXCL8 and TGF-beta 1, respectively. These results provide a potential new strategy for improving MSC-mediated therapy.