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人Tfh细胞体外诱导分化条件的研究
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引用本文:陈语聪,郭薇,雷雯,葛瑶瑶,薛文瑶,李倩文,高向东.人Tfh细胞体外诱导分化条件的研究[J].中国药科大学学报,2017,48(6):733-737.
Cite:CHEN Yucong,GUO Wei,LEI Wen,GE Yaoyao,XUE Wenyao,LI Qianwen,GAO Xiangdong.Optimization of in vitro human follicular helper T cell differentiation condition[J].J China Pharm Univ,2017,48(6):733-737.
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作者单位
陈语聪 中国药科大学生命科学与技术学院分子生物学教研室 
郭薇 中国药科大学生命科学与技术学院分子生物学教研室 
雷雯 中国药科大学生命科学与技术学院分子生物学教研室 
葛瑶瑶 中国药科大学生命科学与技术学院分子生物学教研室 
薛文瑶 中国药科大学生命科学与技术学院分子生物学教研室 
李倩文 中国药科大学生命科学与技术学院分子生物学教研室 
高向东 中国药科大学生命科学与技术学院分子生物学教研室 
基金项目:国家自然科学基金资助项目(No.81573444)
中文摘要:为深入研究滤泡辅助性T细胞(Tfh)的分化调控机制,比较并建立了人Tfh细胞体外诱导分化的模型:分别采用外周血淋巴细胞直接分离以及磁珠分选后的细胞诱导分化,比较不同来源Naï;ve T细胞的分化结果,同时探究了TCR刺激信号anti-hCD3e的固相包被与可溶性刺激对分化效率的影响,并考察极化因子IL-12不同浓度诱导Tfh细胞分化效果,利用流式细胞术检测Tfh分化效率(CD4+CXCR5+ICOS+PD-1+),结合ELISA法检测Tfh标志细胞因子IL-21的表达水平,最终确定了5 μg/mL anti-hCD3e抗体预包被4 ℃过夜,磁珠分选后的T细胞在IL-12终浓度为1 ng/mL时与其他极化因子诱导分化6 d后,分化水平可达20.4%,初步建立了人Tfh细胞体外诱导分化最佳条件,为探讨人Tfh细胞分化机制及相关的靶向Tfh的分子药效、毒性与细胞水平的代谢实验提供了有效的检测平台。
中文关键词:Tfh细胞  TCR信号  体外分化
 
Optimization of in vitro human follicular helper T cell differentiation condition
Abstract:In order to explore the regulation mechanisms of follicular helper T cell(Tfh Cell)differentiation, optimized conditions of in vitro induction from both peripheral blood mononuclear cells and MAC sorted Naï ve CD4+ T cells to human Tfh cells were developed. Induction efficiency difference of TCR signal anti-hCD3e stimulation between coated on solid phase and in soluble phase was also determined. Differentiation efficiency of CD4+CXCR5+ICOS+PD-1+ Tfh cell was determined by FACS while the expression level of IL-21 in cell supernatant was determined by ELISA tests. An ultimate induction condition that 5 μg/mL coated overnight anti-hCD3e stimulated naï ve CD4+ T cells to differentiate into Tfh at an up to 20. 4% percentage was finally determined. The optimization of in vitro induction protocol of human Tfh provided an effective examine platform for the studies on Tfh differentiation mechanisms and related pharmacology, toxicity and metabolic experiments.
keywords:follicular helper T cell  TCR signal  in vitro differentiation
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