SLE患者外周血CD4+T细胞IL-2受体β、γ链表达缺陷与诱导性调节性T细胞转化形成障碍的相关性
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江苏省病原生物学重点实验室开放课题


The predictive role of abnormal expression of IL-2 receptor β/γ chain on CD4+T cells in peripheral blood of SLE patients on the defective induction of induced regulatory T cells
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    摘要:

    目的:通过检测系统性红斑狼疮(SLE)患者外周血CD4+CD25-T细胞及其经体外诱导转化所得CD4+CD25+T细胞表面白细胞介素2受体(IL-2R)β和γ链(CD122和CD132)的表达,探讨其对Foxp3+诱导性调节性T细胞(iTreg)转化形成的影响。方法:分离健康对照和SLE患者外周血单个核细胞(PBMCs)并进一步分离出CD4+CD25-T细胞,分别用流式细胞术和实时定量PCR技术分析CD25-CD122+/CD4+-CD25-CD132+/CD4+比值以及CD4+CD25-T细胞内CD122-CD132(IL-2R β-γ链)mRNA相对表达水平。将CD4+CD25-T细胞诱导转化,进一步分析转化后细胞CD25+CD122+/CD4+-CD25+CD132+/CD4+-CD25+Foxp3+/CD4+-pStat5+/CD4+CD25+的比值。结果:与正常人相比,SLE患者外周血细胞CD4+CD25-T细胞CD122表达变化不明显,但其CD25-CD132+/CD4+比值偏低且与系统性红斑狼疮疾病活动性指数(SLEDAI)呈负相关;CD4+CD25-T细胞内CD132 mRNA相对表达水平亦低于正常对照组;SLE患者外周血CD4+CD25-T细胞经诱导转化后CD4+T细胞中CD25+Foxp3+T细胞亚群比例低于正常对照组,且与SLEDAI呈负相关;SLE患者CD4+CD25-T细胞转化后CD25+CD122+/CD4+-CD25+CD132+/CD4+比例低于对照组,但仅后者与SLEDAI呈负相关;SLE患者细胞转化后胞内磷酸化Stat5表达水平低于对照组,且pStat5+/CD4+CD25+比值与SLEDAI负相关,与CD25+Foxp3+/CD4+比值呈正相关。结论:SLE患者外周血CD4+CD25-T细胞存在明显的CD132(IL-2Rγ链)表达缺陷,且与疾病活动性相关;在其向CD4+CD25+细胞亚群转化过程中同样存在CD132及CD122(IL-2Rβ链)表达缺陷,但只有CD132表达缺陷与疾病活动性-Foxp3分子表达相关,并伴有Stat5磷酸化水平降低,提示IL-2Rγ链表达缺陷及其下游信号减弱与SLE Foxp3+iTreg转化形成障碍密切相关。

    Abstract:

    Objective:To detect the expression of IL-2 receptor β and γ chain(CD122 and CD132) on CD4+CD25-T cells in peripheral blood of systemic tupus erythematosus (SLE)patients and CD4+CD25+T cells which were induced from the former in vitro,as well as assess its effects on the induction of Foxp3+ induced regulatory T cell (iTreg). Methods:Peripheral blood mononuclear cells (PBMCs) were isolated from SLE patients and healthy donors and further isolated CD4+CD25-T cells. The ratio of CD25-CD122+/CD4+,CD25-CD132+/CD4+ and CD122/CD132 (IL-2 receptor β and γ chain) mRNA relative abundance in CD4+CD25-T cells were analyzed by flow cytometry and real-time quantitative PCR,respectively. CD4+CD25-T cells were induced to further analyze the ratio of CD25+CD122+/CD4+,CD25+CD132+/CD4+,CD25+Foxp3+/CD4+,pStat5+/CD4+CD25+. Results:Compared with the normal subjects,there were no significant changes in the expression of CD122 on CD4+CD25-T cells in SLE patients PBMCs,but the ratio of CD25-CD132+/CD4+ was lower and negatively correlated with the SLEDAI;The CD132 mRNA relative abundance in CD4+CD25-T cells was also lower than that in the control group;After CD4+CD25-T cells in peripheral blood of SLE patients were induced,the percentage of CD25+Foxp3+T cells in CD4+T cells was lower than that in the control group and negatively correlated with the SLEDAI;The ratio of CD25+CD122+/CD4+and CD25+CD132+/CD4+ in cells which were transformed from CD4+CD25-T cells in SLE patients was lower than that in the control group,but only the latter was negatively correlated with SLEDAI. The expression of phosphorylated Stat5 in transformed cells of SLE patients was lower than that in the control group and the ratio of pStat5+/CD25+CD4+ was negatively correlated with SLEDAI and positively correlated with the ratio of CD25+Foxp3+/CD4+. Conclusion:There was obviously defective expression of CD132 (IL-2Rγ chain) on CD4+CD25-T cells in peripheral blood of SLE patients and it was correlated with disease activity;The expression of CD132 and CD122 (IL-2Rβ chain) was also defective when the cells were transformed into CD4+CD25+T cell subsets,but only the defective expression of CD132 was correlated with disease activity and Foxp3 expression and was accompanied by lower levels of phosphorylated Stat5,suggesting that the defective expression of IL-2Rγ chain and its weaker downstream signal are closely related with SLE patients Foxp3+iTreg induction obstacle.

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周小斌,杨晓帆,徐安琪,王慧娟,季晓辉. SLE患者外周血CD4+T细胞IL-2受体β、γ链表达缺陷与诱导性调节性T细胞转化形成障碍的相关性[J].南京医科大学学报(自然科学版),2015,(4):455-463

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  • 收稿日期:2014-12-16
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  • 在线发布日期: 2015-05-05
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