小鼠Th17细胞体外诱导分化条件的研究  

Study on Mice Th17 Cell Differentiation Induced in vitro

作　　者：郭薇[1] 王辰[1] 罗成[1] 王宇恒[1] 王欣[1] 高向东[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2013年第6期514-517,共4页Pharmaceutical Biotechnology

基　　金：国家自然科学基金青年基金(No.81202450);中国药科大学中央高校基本科研业务费专项资金资助(No.JKPZ2013013)

摘　　要：为筛选并建立小鼠体外有效的Th17分化,比较并建立3个小鼠Th17细胞体外分化模型:ICR移植排斥Th17细胞分化模型,MOG抗原肽诱导Th17细胞分化模型以及非抗原特异性Th17细胞分化模型,ELISA法分别检测细胞上清炎性因子IL-17的表达水平,软件分析统计学意义,在此基础上进一步优化Th17细胞分化相关影响条件,流式细胞仪检测Th17细胞(CD4+IL17+)的分化水平从诱导前0.58%增加到8.28%,确立小鼠Th17细胞体外分化最佳方案,为研究小鼠Th17细胞分化机制以及相关自身免疫性疾病治疗靶向分子筛选提供有效的检测平台。In order to establish the best differentiation protocol of Thl7 cells in mice in vitro ,3 differentiation models of Th17 cells were established including ICR transplant rejection model, MOG35-55 antigen peptide induced model and cytokine stimulation（ IL-6, TGF-β, IL-23 ）model. To ascertain the best scheme, the level of IL-17 in cell culture supernatant was detected by using correspond- ing ELISA kit. Furthermore,cells surface was stained with mAb against CD4 and intracellular stained with IL17A antibody respec- tively. And then the flowcytometric analysis of CD4 ＋ IL17A ＋ T ceils were measured on a FACS Calibur instrument and results were analyzed using Cell Quest software. Differentiation efficiency of control group is 0.58% and induced group is 8.28%. This study provides effective detection platform for the research of Thl7 cell in vitro activity and Thl7 targeting therapeutic drugs for autoim- mune disease treatment.

关 键 词：TH17细胞 免疫排斥 体外分化 

分 类 号：R392.12[医药卫生—免疫学]