上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (11): 1380-.doi: 10.3969/j.issn.1674-8115.2010.11.015

• 论著（临床研究） • 上一篇    下一篇

成人微小病变肾病患者外周血CD4+Foxp3+调节性T细胞的研究

林芙君1, 单剑萍1, 朱 淳1, 陆 玮1, 潘秀军2, 袁向亮2, 蒋更如1   

  1. 上海交通大学 |医学院附属新华医院 1.肾脏内科, 2. 检验科, 上海 200092
  • 出版日期:2010-11-25 发布日期:2010-11-29
  • 通讯作者: 蒋更如, 电子信箱: jianggengru@hotmail.com。
  • 作者简介:林芙君（1982—）, 女, 住院医师, 硕士；电子信箱: seatoneal@hotmail.com。

Study of CD4+Foxp3+ regulatory T cells in peripheral blood of adult minimal change disease

LIN Fu-jun1, SHAN Jian-ping1, ZHU Chun1, LU Wei1, PAN Xiu-jun2, YUAN Xiang-liang2, JIANG Geng-ru1   

  1. 1.Department of Nephrology, 2.Department of Clinical Laboratory, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China
  • Online:2010-11-25 Published:2010-11-29

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摘要：

目的 研究CD4+Foxp3+调节性T细胞（CD4+Foxp3+Treg）及亚群在成人微小病变肾病（MCD）患者外周血CD4+T细胞中的分布，探讨CD4+Foxp3+Treg参与MCD发病的可能机制。方法 以27例经肾穿刺活检确诊为MCD的初发成人患者作为研究对象，采用Foxp3-FITC/CD45RA-PE/CD3-ECD/CD4-PC7抗体组合经流式细胞仪检测外周血单个核细胞（PBMCs）中CD4+Foxp3+Treg及亚群占CD4+T细胞的百分比。Real-Time PCR检测PBMCs中Foxp3、Th17转录因子RORc以及细胞因子TGF-β1、IL-6、IL-17A 、IL-23 mRNA表达。设立正常对照。结果 流式细胞检测显示CD4+Foxp3+Treg分成3个细胞群体，分别为CD45RA+Foxp3low(Ⅰ区，静息期Treg)、CD45RA-Foxp3high（Ⅱ区，活化Treg)和CD45RA-Foxp3low(Ⅲ区)。MCD组Ⅰ区+Ⅱ区细胞和Ⅱ区细胞占CD4+T细胞的百分比显著低于对照组（P<0.01）。MCD组PBMCs中Foxp3 mRNA表达较对照组下调（P<0.05），RORc mRNA表达较对照组上调（P<0.05），IL-6、IL-17A、IL-23 mRNA表达均显著高于对照组（P<0.05和P<0.01）。相关性分析显示，MCD组IL-17A mRNA表达与Ⅱ区细胞占CD4+T细胞的百分比呈显著负相关（r=－0.81，P<0.05）。结论 成人MCD患者外周血Treg及其活化功能亚群减少，同时伴有Th17转录基因及其相关细胞因子表达的异常升高。提示Treg Foxp3 mRNA表达下调及Treg/Th17细胞比例失衡可能与MCD的发病有关。

关键词: 微小病变肾病, 成人, 调节性T细胞, Foxp3

Abstract:

Objective To observe the distribution of CD4+Foxp3+ regulatory T cells (CD4+Foxp3+ Treg) and the subsets in CD4+T cells in peripheral blood of adults with minimal change disease (MCD), and explore the significance of CD4+Foxp3+Treg in the pathogenesis of MCD. Methods Twenty-seven adults with MCD initially diagnosed by renal biopsy were selected. The antibody combination of Foxp3-FITC/CD45RA-PE/CD3-ECD/CD4-PC7 was adopted and flow cytometry was employed to determine the percentage of CD4+Foxp3+ Treg and the subsets in CD4+T cells in peripheral mononuclear cells (PBMCs). The expression of Foxp3, RORc, TGF-β1, IL-6, IL-17A and IL-23 mRNA in PBMCs was detected by Real-Time PCR. Besides, normal controls were established. Results Flow cytometry revealed that CD4+Foxp3+Treg were divided into three subsets: CD45RA+Foxp3lowcells(Fr.Ⅰ, resting Treg), CD45RA-Foxp3highcells (Fr.Ⅱ, active Treg) and CD45RA-Foxp3lowcells (Fr.Ⅲ). The percentages of cells of Fr.Ⅰ+Fr.Ⅱ and cells of Fr.Ⅱ in CD4+ T cells in MCD group were significantly lower than those in control group (P<0.01). The expression of Foxp3 mRNA in PBMCs in MCD group was significantly lower than that in control group （P<0.05）, while the expression of RORc mRNA in PBMCs in MCD group was significantly higher than that in control group （P<0.05）, and the expression of IL-6, IL-17A and IL-23 mRNA was significantly higher than that in control group （P<0.05, P<0.01）. Correlation analysis revealed that the expression of IL-17A mRNA was negatively related to the percentage of cells of Fr.II in CD4+T cells in MCD group （r=-0.81，P<0.05）. Conclusion In adults with MCD, there is decreased expression of Treg and active Treg subsets and increased expression of Th17-related gene and cytokines in peripheral blood, which suggests that the down-regulated expression of Treg Foxp3 mRNA and Treg/Th17 imbalance might be involved in the pathogenesis of MCD.

Key words: minimal change disease, adult, regulatory T cells, Foxp3