体外高效诱导人MDSC分化和扩增的方法探索  

Development of a method to efficiently induce human MDSCs differentiation and expansion in vitro

作　　者：周鑫阳 朱晓露 祁荆荆 刘畅[3] ZHOU Xin-yang;ZHU Xiao-lu;QI Jing-jing;LIU Chang(Graduate School,Dalian Medical University,Dalian 116044,China;Department of Immunology,College of Basic Medical Science,Dalian Medical University,Dalian 116044,China;Department of Rheumatology and Immunology,Dalian Municipal Central Hospital,Dalian 116089,China)

机构地区：[1]大连医科大学研究生院,大连116044 [2]大连医科大学基础医学院免疫学教研室,大连116044 [3]大连市中心医院风湿免疫科,大连116089

出　　处：《现代免疫学》2024年第1期45-50,共6页Current Immunology

基　　金：国家自然科学基金青年项目(82201990);大连市科技人才,青年科技之星(2022RQ070);辽宁省教育厅自然科学青年科技人才“育苗”项目(LZ2020051);辽宁省博士科研启动计划项目(2020-BS-191)。

摘　　要：为探索体外稳定、高效诱导人髓源抑制性细胞(myeloid-derived suppressor cell,MDSC)分化与扩增的实验方法,为体外研究人MDSC功能及作用机制提供新的实验技术支持,收集健康人全血后采用Ficoll密度梯度离心法分离PBMC,贴壁法纯化单核细胞。用人GM-CSF和IL-6(各10、20、40和80 ng/mL)刺激PBMC或单核细胞4、7、14和21 d,与PBMC共培养3 d;采用FACS检测CD33^(+)MDSC、T细胞增殖和CD3^(+)IFN-γ^(+)T细胞比例。结果显示,常规方法仅诱导PBMC或单核细胞分化为较低比例的CD33^(+)MDSC;改善培养方法,用人GM-CSF和IL-6(各80 ng/mL)刺激单核细胞14 d(每4~5天换1次液),可诱导出较高比例的CD33^(+)MDSC,且CD33^(+)MDSC表现出显著抑制T细胞增殖及分泌IFN-γ的功能。该研究提示,改良后的培养方法可高效诱导人单核细胞分化成为CD33^(+)MDSC,CD33^(+)MDSC具有较强的免疫抑制功能。This study aims to develop a method to efficiently and stably induce the differentiation and expansion of human myeloid-derived suppressor cells(MDSC)in vitro and provide new experimental technology for the study of the function and mechanism of human MDSC.For this purpose,PBMC of healthy volunteers were isolated with Ficoll density gradient centrifugation.Next,monocytes were separated by anchoring technique.PBMC or monocytes were stimulated with GM-CSF and IL-6(10,20,40,80 ng/mL each)for 4,7,14,and 21 days before co-culturing with PBMC for 3 days.The proliferation of CD33^(+)MDSC and T cells and the proportion of CD3^(+)IFN-γ^(+)T cells were detected by FACS.The results showed that the PBMC or monocytes were induced to differentiate to less CD33^(+)MDSC by conventional methods.However,by improved methods,the monocytes cultured with GM-CSF and IL-6(each 80 ng/mL)for 14 days(refreshing the culture solution every 4-5 days)were induced to differentiate more efficiently into a higher proportion of CD33^(+)MDSC,which showed significant inhibition of T cell proliferation and IFN-γsecretion.This study suggests that with the improved method,the monocytes are efficiently induced to differentiate into CD33^(+)MDSC,which has significant suppressive function.

关 键 词：人髓源抑制性细胞 体外分化 实验技术 

分 类 号：R392-33[医药卫生—免疫学]