AIRE通过调控Notch1表达影响CD4^+CD25^+Treg的作用及其机制  

Effect of AIRE on CD4^+CD25^+Treg by regulating Notch1 expression and mechanism

作　　者：吴静[1] 杨巍[1] 孙际童[1] 苏静波[1] 魏晓曦[1] 李一[1] 

机构地区：[1]吉林大学基础医学院免疫学教研室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2009年第6期983-986,共4页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金面上项目资助课题(30872307)

摘　　要：目的:研究AIRE基因是否可通过调控Notch1的表达而影响CD4+CD25+Treg细胞的诱导及功能,探讨AIRE在外周免疫耐受中的作用机制。方法:将RAW264.7细胞分为转染组和未转染组,采用脂质体法将pEGFPC3-AIRE质粒转入RAW264.7细胞,采用RT-PCR法检测各组AIRE mRNA的表达及Notch1 mRNA表达的变化。将RAW264.7细胞分为转染组、转染并经Notch1抗体阻断组、阴性对照组及未转染并经Notch1抗体阻断组,分别采用FACS法及RT-PCR法检测各组RAW264.7细胞对CD4+CD25+Treg细胞数量及Foxp3 mRNA表达的影响。结果:质粒成功转入RAW264.7细胞内。AIRE转染组较未转染组Notch1 mRNA表达增加。AIRE转染组较未转染组CD4+CD25+Treg细胞的数量及功能相关基因Foxp3 mRNA的表达水平均增加,经Notch1抗体阻断后,AIRE转染组与未转染组CD4+CD25+Treg细胞的数量及功能均较对照组降低。结论:AIRE可能通过上调RAW264.7细胞上Notch1的表达进而影响CD4+CD25+Treg细胞的诱导产生及功能,从而在维持外周免疫耐受方面发挥一定的作用。Objective To study the function of AIRE on inducement and function of the CD4^＋CD25^＋Treg cells by regulating Notch1 expression,and discuss the mechanism of AIRE in peripheral immune tolerance.Methods RAW264.7 cells were divided into transfected group and untransfected group,PEGFPC3-AIRE plasmid was transfected with liposome,RT-PCR was used to detect the expressions of AIRE mRNA and Notch1 mRNA in every group.RAW264.7 cells were divided into transfected group,untransfected group,transfected and anti-Notch1 blocking group,untransfected and anti-Notch1 blocking group;FACS and RT-PCR were used to detect the number of CD4^＋CD25^＋Treg cells and the expression of Foxp3 mRNA in every group.Results AIRE gene was successfully transfected into the RAW264.7 cells.Notch1 mRNA expression in transfected group was significantly higher than that in untransfected group.The number of CD4^＋CD25^＋Treg cells and Foxp3 mRNA expression in transfected group were significantly higher than that in untransfected group,after applying the Notch1 antibody,compared with control group,the number of CD4^＋CD25^＋Treg cells and Foxp3 mRNA expression were lower in experimental group.Conclusion AIRE could influence the production and function of CD4^＋CD25^＋Treg cells by up-regulation of Notch1 for keeping the function on peripheral immune tolerance.

关 键 词：自身免疫调节因子 NOTCH1 CD4+CD25+TREG细胞 

分 类 号：R392[医药卫生—免疫学]