人脐带间充质干细胞外泌体免疫调节功能的研究  

Study of immunomodulatory function of exosomes derived from human umbilical cord mesenchymal stem cells

作　　者：刘明[1] 汪劲松 刘沐芸 胡祥 徐军[1] 

机构地区：[1]呼吸疾病国家重点实验室,广州呼吸疾病研究所,广州医科大学附属第一医院,广州510120 [2]深圳市北科生物科技有限公司

出　　处：《中华医学杂志》2015年第32期2630-2633,共4页National Medical Journal of China

基　　金：国家自然科学基金（81200050）;深圳市科技研发资金（CXY201107010176A）;广州市属高校科研计划（2012C157）

摘　　要：目的 探讨人脐带间充质干细胞（hUC-MSCs）分泌的外泌体（exosomes）的免疫调节能力.方法 分离培养hUC-MSCs,收集第3代hUC-MSCs上清,提取外泌体,Western印迹检测其表面特异标志CD81、CD9的表达,BCA法测定浓度;不同浓度外泌体与CD3/CD28单抗刺激的正常人外周血单个核细胞（PBMC）作用72 h,流式检测作用前后Th17和Treg细胞的比例变化,检测对CD4^＋T、CD8^＋T细胞增殖能力的影响,ELISA检测干扰素（IFN）-γ、白细胞介素（IL）-6、肿瘤坏死因子α（TNF-α）、转化生长因子β1（TGF-β1）的水平变化.结果 hUC-MSCs外泌体高表达CD9、CD81,可明显抑制CD4＋T和CD8＋T细胞的增殖,升高CD4^＋ CD25^＋ FoxP3^＋ Treg细胞比例,经CD3/CD28单抗刺激后CD45＋ CD4^＋ Ki67^＋和CD45^＋ CD8^＋ Ki67^＋细胞比例为85.3％±5.6％和72.6％±6.3％,同时升高PBMC分泌的TGF-β1水平,降低IFN-γ、IL-6、TNF-α的水平（均P＜0.05）.结论 hUC-MSCs分泌的外泌体具有免疫调节功能,有望成为一种免疫治疗的新选择.Objective To investigate the immumodulation ability of exosomes secreted from human umbilical cord-derived mesenchymal stem cells （hUC-MSCs）.Methods hUC-MSCs were isolated and cultured.Exosomes were isolated from the culture media of the third-generation hUC-MSCs.The expression of specific surface marker CD9 and CD81 were detected by Western blot,and the concentration of hUC-MSCs exosomes（hUC-MSCs-ex） was evaluated by BCA assay.CD3/CD28-stimulated peripheral blood mononuclear cells（PBMCs） from healthy donor were co-cultured with different concentration of hUC-MSCs-ex for 72 h.The percentage of Th17 and Treg cells and the proliferation of CD4^＋ and CD8^＋ T cells were detected by flow cytometry.ELISA was used to test the level of IFN-γ,IL-6,TNF-α and TGF-β1.Results hUC-MSCs-ex inhibited the proliferation of CD4 ^＋ and CD8 ^＋ cells obviously,and increased the proportion of CD4 ^＋ CD25^ ＋ FoxP3 ^＋ Treg cells,with high expression of CD81 and CD9.After CD3/CD28 monoclonal antibody stimulated,the percentage of CD45 ^＋ CD4^ ＋ Ki67 ^＋ and CD45 ^＋ CD8 ^＋ Ki67 ^＋ cells were 85.3% ± 5.6% and 72.6% ± 6.3%,respectively.Meanwhile,the level of TGF-β1 were elevated and the level of IFN-γ,IL-6 and TNF-α were decreased （P 〈 0.05）.Conclusion hUC-MSCs-ex has the immunomodulatory functionin vitro,which could be a new therapeutic agent for the treatment of immune disorders.

关 键 词：间充质干细胞 脐带 外泌体 免疫 

分 类 号：R392[医药卫生—免疫学]