摘要
背景:Th22细胞是一种新型CD4^+辅助T细胞,人脐带间充质干细胞外泌体具有T细胞免疫调节功能。目的:探讨人脐带间充质干细胞分泌的外泌体对Th22细胞免疫调节功能的影响。方法:分离培养人脐带间充质干细胞,STR基因分型鉴定细胞株特性。收集第4代人脐带间充质干细胞上清,提取外泌体,透射电镜观察形态与粒径,Western blot检测表面特异标志CD63、CD81的表达,BCA法测定外泌体浓度;人脐带间充质干细胞外泌体与淋巴细胞刺激剂活化的正常人外周血单个核细胞共培养6 h,流式细胞术检测Th22细胞的比例以及对CD3^+CD4^+T、CD3^+CD8^+T细胞增殖能力的影响;共培养72 h,流式细胞术检测白细胞介素22的含量。结果与结论:成功建立人脐带间充质干细胞原代细胞系且可稳定传代。人脐带间充质干细胞外泌体具有典型的外泌体形态及标识性蛋白,可明显抑制CD3^+CD4^+T细胞(P<0.05)和CD3^+CD8^+T细胞(P<0.01)的增殖,抑制Th22细胞的表达比例并降低其主要细胞因子白细胞介素22的表达含量(P<0.01)。结果表明,人脐带间充质干细胞分泌的外泌体可以抑制Th22细胞的表达及功能,具有免疫调节能力,有望成为一种免疫治疗的新选择。
BACKGROUND:Th22 cells are a novel type of CD4+helper T cells.Exosomes derived from human umbilical cord mesenchymal stem cells(hUCMSCs)have T cell immunomodulatory function.OBJECTIVE:To investigate the Th22 cell immunomodulatory ability of exosomes secreted from hUCMSCs.METHODS:hUCMSCs were isolated and cultured,and cell line was authenticated using short tandem repeat(STR)analysis.Exosomes were extracted from the supernatant of passage 4 hUCMSCs.Morphology and particle size of the exosomes were detected under transmission electron microscope.The expression of specific surface marker CD63 and CD81 were detected by western blot.The concentration of exosomes was evaluated by BCA assay.Cell activation cocktail stimulated peripheral blood mononuclear cells from healthy donors were co-cultured with exosomes for 6 hours.The percentage of Th22 cells and the proliferation of CD3+CD4+T cells and CD3+CD8+T cells were detected by flow cytometry.Flow cytometry was also used to test the level of interleukin 22 at 72 hours of co-culture.RESULTS AND CONCLUSION:Successfully established hUCMSCs primary cell line could stably passage.Exosomes secreted from hUCMSCs had typical exosomal morphology and marker proteins,and significantly inhibited the proliferation of CD3+CD4+T cells(P<0.05)and CD3+CD8+T cells(P<0.01).These exosomes also inhibited the proliferation of Th22 cells,and reduced the level of interleukin 22 in the cells(P<0.01).These findings indicate that exosomes secreted from hUCMSCs inhibit Th22 cell expression and function,and have the immunomodulatory function of Th22 cells in vitro,which can be a new therapeutic agent for the treatment of immune disorders.
作者
李伟伟
李晓丰
侯萍
李剑平
Li Wei-wei;Li Xiao-feng;Hou Ping;Li Jian-ping(Shenyang Blood Center(Liaoning Provincial Blood Center),Shenyang 110044,Liaoning Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第29期4657-4662,共6页
Chinese Journal of Tissue Engineering Research
基金
2017年辽宁省博士科研启动基金指导计划项目(21070520339)~~
