人脐带间充质干细胞来源外泌体对Treg和TH17细胞的调节作用  

Regulatory Effect of Exosomes Derived from Human Umbiilcal Cord Mesenchymal Stem Cells on Treg and TH17 Cells

作　　者：郭礼妍 赖沛龙 耿素霞 陈晓梅 王玉连 黄恬 杜欣 翁建宇 GUO Li-Yan;LAI Pei-Long;GENG Su-Xia;CHEN Xiao-Mei;WANG Yu-Lian;HUANG Tian;DU Xin;WENG Jian-Yu(School of Medicine,South China University of Technology,Guangzhou 510006,Guangdong Province,China;Department of Hematology,Guangdong Provincial People Hospital,Guangdong Provincial Academy of Medical Sciences,Guangdong Elderly Research Institute,Guangzhou 510080,Guangdong Province,China)

机构地区：[1]华南理工大学医学院,广东广州510006 [2]广东省人民医院广东省医学科学院广东省老年医学研究所,广东广州510000

出　　处：《中国实验血液学杂志》2019年第1期221-226,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(81671585);广东省科技计划项目(2017B020230004);(2015B020227003);(201614)

摘　　要：目的:探讨人脐带间充质干细胞来源外泌体对Treg和TH17细胞的影响。方法:应用超高速离心法获取脐带间充质干细胞培养上清来源的外泌体,通过电子显微镜检、Nanosight和Western blot方法对外泌体形态、粒径和所含蛋白进行鉴定。在外周血单个核细胞(PBMC)的培养体系中,以外泌体共培养为实验组,PBS组为对照组,应用流式细胞术检测实验组与对照组Treg及TH17细胞比例。结果:人脐带间充质干细胞来源的外泌体在电子显微镜下呈杯状圆盘形,平均粒径142 nm,表达外泌体标志性蛋白CD9及CD63。流式细胞术结果显示,外泌体组Treg(CD4^+CD25^+Foxp3^+)细胞比例明显高于无外泌体组(5.30±0.13%vs 3.38±0.09%)(P<0.001),而外泌体组TH17(CD4^+IL17A^+)细胞比例明显低于无外泌体组(7.78±0.31%vs 11.33±0.24%)(P<0.001)。结论:人脐带间充质干细胞来源的外泌体能通过上调Treg和下调TH17参与免疫调节,提示有望成为新的"非细胞"免疫调节制剂。Objective: To investigate the effects of exosomes from human umbilical cord mesenchymal stem cells on the development of Treg and TH17 cells. Methods: Exosomes from the serum-free-culture supernatants of hUCMSC were harvested by ultracentrifugation. The electron microscopy, nanoparticle tracking analysis and western blot were used to identify the hUC-MSC-exosomes, such as the morphology, the paticle chameter, and the protein content. The PBMC stimulated with anti-CD3/CD28 were incubated with the exosomes for five days, and then the percentage changes of Treg and TH17 cells were analyzed by using flow cytometry. Results: The hUC MSC-derived exosomes were saucer-like in morphology the averge diameter was approximately 142 nm. They were identified as positive for CD9 and CD63. Flow cytometry showed that the proportion of CD4^+CD25^+Foxp3^+ Treg cells in the PBMC were significantly higher, but the proportion of CD4^+IL17A^+ T cells in the hUC-MSC-exosome group was obviously lower than that in the group without the hUC-MSC-exosom(control group)(P <0.05). Conclusion: The hUC-MSC-exosomes have an immunomodulatory effect on T cells in vitro by increasing the ratio of Treg and reducing the ratio of TH17 cells, expecting the hUC-MSC-exosom as a novel cell-free target for immunotherapy.

关 键 词：人脐带间充质干细胞 外泌体 免疫调节 调节性T细胞 TH17细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]