中国药理学通报2017，Vol.33Issue(1)：45-51,7.DOI:10.3969/j.issn.1001-1978.2017.01.009
干扰素γ刺激hUC-MS Cs分泌外泌体促进调节性T细胞生成
Exosomes secreted from IFN-γprestimulated hUC-MSCs induce regulatory T cells
摘要
Abstract
Aim To investigate whether human umbili-cal cord mesenchymal stem cells(hUC-MSCs)exposed to inflammatory conditions could release large amounts of exosomes to induce regulatory T cells(Treg).Meth-ods hUC-MSCs were isolated by enzyme digestion method.(In vitro)interferonγ(IFN-γ)was added in-to hUC-MSCs to mimic inflammatory microenviron-ments,then exosomes were extracted from the superna-tant of normal conditional medium or IFN-γpretreated hUC-MSCs.Both sources of exosomes,Nor-hUC-exo and IFN-γ-stimulated hUC-exo, were identified by Nanoparticle Trafficking Analysis (NTA )and Western blot for the exosome-enriched protein CD63 .Next,hu-man peripheral blood mononuclear cells (PBMCs ) stimulated with PHA were respectively co-cultured with hUC-MSCs,IFN-γ-pretreated-hUC-MSCs,hUC-MSCs exosomes or IFN-γ-stimulated-hUC-MSCs exosomes for 5 days to assess the exosomes-T cells communication. The proliferation rate of PBMCs and frequency of CD4 +/CD25 +/Foxp3 + Treg were measured by flow cytometry.Results The isolated cells from human um-bilical cord tissue,which were positive for CD73, CD44,CD29,CD90 and HLA-ABC,but were nega-tive for CD31 and CD34,were mesenchymal stem cells indeed.After IFN-γtreatment,hUC-MSCs secreted nu-merous exosomes(P<0.05 ).Morerover,there was a significantly higher level of CD63 ,but no difference in diameter between Nor-hUC-exo and IFN-γ-stimulated hUC-exo.IFN-γ-stimulated hUC-exo had a superior a-bility compared with Nor-hUC-exo to suppress the pro-liferation of PHA stimulated PBMCs due to their upreg-ulation of the percentage of Treg (1 1.53 ±0.88% vs 6.60 ±0.56%,P <0.01 ).Conclusion hUC-MSCs could promote the expression of Treg to modulate im-munosuppression through exosomes,especially for IFN-γ-licenced exosomes,which might carry much immu-notherapeutic potential.关键词
脐带/间充质干细胞/外泌体/炎性微环境/免疫调节/调节性T细胞Key words
umbilical cord/mesenchymal stem cells/exosomes/inflammatory microenvironment/immunodu-lation/regulatory T cells引用本文复制引用
杨翔宇,李晓红,肖静,胡洁媚,冯娟,霍然,何国东,吴岳恒,余细勇..干扰素γ刺激hUC-MS Cs分泌外泌体促进调节性T细胞生成[J].中国药理学通报,2017,33(1):45-51,7.基金项目
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（No 2015B020225006）；广东省科技计划国际合作项目 （）
