干扰素γ刺激hUC-MSCs分泌外泌体促进调节性T细胞生成

发布时间：2018-05-10 14:32

本文选题：脐带 + 间充质干细胞　；参考：《中国药理学通报》2017年01期

【摘要】：目的探讨生理和炎性微环境下人脐带间充质干细胞(human umbilical cord derived mesenchymal stem cells,h UCMSCs)能否通过外泌体诱导调节性T细胞的生成来发挥免疫抑制功能。方法胶原酶消化法分离h UC-MSCs,应用流式细胞术鉴定h UC-MSCs。IFN-γ模拟体内炎性微环境,以未预处理为对照,分别提取外泌体,得到Nor-h UC-exo和IFN-γ-stimulated h UC-exo,分析其浓度及粒径分布等特征、并鉴定表面标记蛋白CD63的表达。采用h UC-MSCs、Nor-h UCexo、IFN-γpretreated-h UC-MSCs、IFN-γstimulated h UC-exo与人外周血单个核细胞共培养5 d后,流式细胞术检测T细胞的增殖、调节性T细胞的比例变化。结果 h UC-MSCs高表达CD73、CD44等间充质干细胞标志物。IFN-γ刺激前后外泌体粒径无明显变化,但IFN-γ刺激后分泌量和CD63增加(P0.05)。CFSE染料示踪结果显示,h UC-MSCs来源的外泌体抑制PBMCs增殖(P0.01),且IFN-γ刺激后明显提高外泌体的免疫抑制能力(P0.01)。在活化T细胞中,IFN-γ-stimulated h UC-exo组Treg比例(11.53±0.88%)与IFN-γpretreated-h UC-MSCs组(7.54±0.50%)(P0.05)、Norh UC-exo组(6.60±0.56%)(P0.01)、对照组(3.87±0.73%)(P0.01)相比升高。结论 h UC-MSCs可通过分泌外泌体来发挥免疫调节作用,在炎症因子刺激下h UCMSCs分泌的外泌体明显促进调节性T细胞的生成,h UC-exo可能是潜在的免疫抑制载体。
[Abstract]:Objective to investigate whether human umbilical cord derived mesenchymal stem cells can induce regulatory T cell formation in human umbilical cord mesenchymal stem cells (UCMSCs) in physiological and inflammatory microenvironment to play an immunosuppressive role. Methods the hUC-MSCswere isolated by collagenase digestion. The inflammatory microenvironment of hUC-MSCs.IFN- 纬 was identified by flow cytometry. The exocrine was extracted from the unpretreated control, and the Nor-h UC-exo and IFN- 纬 -stimulated UC-exo were obtained, and their concentration and particle size distribution were analyzed. The expression of surface marker protein CD63 was identified. Human peripheral blood mononuclear cells (PBMC) were co-cultured for 5 days with hUC-MSCsCoculture of IFN- 纬 pretreated-h stimulated h UC-exo. The proliferation of T cells and the percentage of regulatory T cells were detected by flow cytometry. Results h UC-MSCs overexpression of CD73, CD44 and other mesenchymal stem cell markers, such as. IFN- 纬, had no significant change in the size of exocrine. But after IFN- 纬 stimulation, the secretion and CD63 increased P0.05N 路CFSE dye tracer. The results showed that exosomes derived from UC-MSCs inhibited the proliferation of PBMCs P0.01, and IFN- 纬 increased the immunosuppressive ability of exocrine. The ratio of Treg in IFN- 纬 -stimulated h UC-exo group (11.53 卤0.88) was significantly higher than that in IFN- 纬 pretreated-h UC-MSCs group (7.54 卤0.50) and the control group (3.87 卤0.73). Conclusion h UC-MSCs can play an immunomodulatory role by secreting exosomes, and the secretion of h UCMSCs stimulated by inflammatory cytokines can obviously promote the formation of regulatory T cells. H UC-exo may be a potential immunosuppressive vector.
【作者单位】：南方医科大学药学院;广东省人民医院医学研究中心广东省医学科学院;广东食品药品职业学院国际交流学院;广州医科大学药学院;
【基金】：国家自然科学基金资助项目(No 81330007,81120108003) 广东省科技计划重点项目(No2015B020225006);广东省科技计划国际合作项目(No2014A05053047)
【分类号】：R392

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