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

J4 ›› 2009, Vol. 35 ›› Issue (6): 983-986.

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

吴静, 杨巍, 孙际童, 苏静波, 魏晓曦, 李一

（吉林大学基础医学院免疫学教研室|吉林长春 130021）

收稿日期:2009-07-24 出版日期:2009-11-28 发布日期:2009-11-28
通讯作者: 李一 E-mail:Tel:0431-85619476，E-mail:yilili19@yahoo.com.cn
作者简介:吴静（1980-）|女|吉林省白山市人|在读医学博士|主要从事免疫耐受机制研究。
基金资助:
国家自然科学基金面上项目资助课题（30872307）

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

TUN Jing, YANG Wei, SUN Ji-Tong, SU Jing-Bei, WEI Xiao-Xi, LI Yi

(Department of Immunology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China)

Received:2009-07-24 Online:2009-11-28 Published:2009-11-28

摘要/Abstract

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。

关键词: 自身免疫调节因子；Notch1；CD4+CD25+Treg细胞

Abstract:

Abstract: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.

Key words: autoimmune regulator；Notch1；CD4+CD25+Treg cells

中图分类号:

R392

吴静, 杨巍, 孙际童, 苏静波, 魏晓曦, 李一. AIRE通过调控Notch1表达影响CD4+CD25+Treg的作用及其机制[J]. J4, 2009, 35(6): 983-986.

TUN Jing, YANG Wei, SUN Ji-Tong, SU Jing-Bei, WEI Xiao-Xi, LI Yi. Effect of AIRE on CD4+CD25+Treg by regulating Notch1 expression and mechanism[J]. J4, 2009, 35(6): 983-986.