A20在调节性T细胞IL-2信号通路中的作用研究*

doi:10.3969/j.issn.1671-2587.2022.04.011

临床输血与检验 ›› 2022, Vol. 24 ›› Issue (4): 464-468.DOI: 10.3969/j.issn.1671-2587.2022.04.011

A20在调节性T细胞IL-2信号通路中的作用研究^*

陈富强, 曲莉莉

250014 济南,山东省血液中心（陈富强）; 山东大学齐鲁医院（曲莉莉）

收稿日期:2022-04-18 出版日期:2022-08-20 发布日期:2022-08-19
通讯作者: 曲莉莉,主要从事影像医学与核医学研究,（E-mail）lili.qu@sdu.edu.cn。
作者简介:陈富强（1983-）,男,副主任技师,硕士,主要从事血液免疫学和输血传染病检测工作,（E-mail）cfqcfqcfq@163.com。
基金资助:
*本课题受山东省医药卫生科技发展计划项目（No.2019WS537）资助

A Role of A20 in the Signaling Pathway of Interleukin-2 in Regulatory T Cells

CHEN Fu-qiang, QU Li-li

Blood Center of Shandong Province, Jinan 250014

Received:2022-04-18 Online:2022-08-20 Published:2022-08-19

摘要/Abstract

摘要： 目的研究A20（ubiquitin E3 ligase A20）对调节性T细胞（regulatory T cell,Treg）中白细胞介素2（IL-2）信号通路的影响。方法将A20^flox/flox小鼠与CD4^-cre小鼠杂交,得到在CD4⁺ T细胞中A20基因条件敲除的CD4^-cre A20^flox/flox小鼠作为实验组,A20^flox/flox小鼠作为对照组。体外实验分别分离实验组和对照组胸腺和脾脏淋巴细胞,流式细胞术检测Treg表面标记分子CTLA-4、CD73、FR4、 GITR表达水平,加入IL-2刺激体外培养的Treg细胞,分别使用pStat5, pERK1/2和pAKT抗体标记,检测磷酸化水平。体内实验尾静脉注射实验组和对照组Treg和正常效应T细胞组合,观察体重变化,探讨不同组Treg的功能变化,尾静脉分别注射5-溴脱氧尿嘧啶核苷（BrdU）追踪Treg的增殖来源,膜联蛋白V （AnmeXin V）标记流式细胞术分析细胞凋亡情况。结果 A20基因条件敲除小鼠的T细胞发育没有明显缺陷,胸腺和外周淋巴组织Treg的比例和数量显著增加,并在体内表现出正常的抑制作用,IL-2信号通路中的Stat5表达明显增强。结论 A20在Treg增殖中起重要负调控作用,主要通过抑制IL-2信号通路中Stat5的磷酸化,抑制IL-2分泌和Treg细胞增殖。

关键词: A20, 调节性T细胞, Foxp3, IL-2, 获得性免疫

Abstract: Objective To study the effect of A20 on the signaling pathway of interleukin-2 （IL-2） in regulatory T cells （Treg）. Methods A20^flox/flox mice were hybridized with CD4^-cre mice to get CD4^-cre A20^flox/flox mice, which, with A20 gene conditionally knocked-out in CD4⁺ T cells, were assigned into the experimental group, and A20^flox/flox mice into the control group. Spleen cells and lymph node cells of the experimental and control groups were then isolated. The proliferation and activity of Treg were detected by flow cytometry and in vivo experimentation. The source of Treg proliferation was traced by BrdU. The expressions of STAT5, ERK1/2 and Akt were identified by staining with anti-pStat5, pERK1/2 and pAKT markers respectively.Results In mice with conditional knockout of A20 gene, T cell development was not significantly defective. The proportion and number of regulatory T cells in the thymus and peripheral lymphoid tissues of A20 cKO mice increased significantly and showed a normal inhibitory effect in vivo. Additionally, Stat5 expression in IL-2 signaling pathway was markedly enhanced.Conclusion A20 could boost the activity of Treg cells by enhancing STAT5 and IL-2 signaling pathway.

Key words: A20, Regulatory T cells, Foxp3, IL2, Adaptive immunity

中图分类号:

R575.1

陈富强, 曲莉莉. A20在调节性T细胞IL-2信号通路中的作用研究^*[J]. 临床输血与检验, 2022, 24(4): 464-468.

CHEN Fu-qiang, QU Li-li. A Role of A20 in the Signaling Pathway of Interleukin-2 in Regulatory T Cells[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2022, 24(4): 464-468.

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A20在调节性T细胞IL-2信号通路中的作用研究^*

A Role of A20 in the Signaling Pathway of Interleukin-2 in Regulatory T Cells

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