[1]何跃,韩容,刘宏,等.Herpes virus entry mediator/CD160介导人调节性T细胞对CD4+效应性T细胞的抑制作用[J].第三军医大学学报,2011,33(10):1012-1015.
 He Yue,Han Rong,Liu Hong,et al.Human regulatory T cells exert inhibitory function on CD4+ effector T cells through interaction between herpes virus entry mediator and CD160[J].J Third Mil Med Univ,2011,33(10):1012-1015.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第10期
页码:
1012-1015
栏目:
论著
出版日期:
2011-05-30

文章信息/Info

Title:
Human regulatory T cells exert inhibitory function on CD4+ effector T cells through interaction between herpes virus entry mediator and CD160
作者:
何跃韩容刘宏吴雄飞
第三军医大学西南医院肾内科
Author(s):
He Yue Han Rong Liu Hong Wu Xiongfei
Department of Nephropathy, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
共刺激分子CD160HVEM调节性T细胞器官移植
Keywords:
costimulator CD160 herpes virus entry mediator regulatory T cell organ transplantation
分类号:
R392.12;R392.4;R392-33
文献标志码:
A
摘要:
目的    研究HVEM、CD160在不同CD4+T细胞亚群,不同功能状态下的表达以及HVEM-CD160通路对CD4+CD25+ 调节性T细胞(regulatory T cell,Treg)免疫调节作用的影响。    方法    采用流式细胞分析术分别检测3例不同个体CD4+CD25+ Tregs及CD4+CD25-  T细胞在1 000 IU/ml人重组IL-2、与细胞数量1∶1的抗CD3/CD28 mAb包被磁珠的刺激下第0、3、5天HVEM或CD160的表达情况。采用1.25、2.5、5 μg/ml HSV1 gD蛋白处理4例不同个体Treg,培养5 d 后,将处理的Treg与Teff在体外以1∶1混合并加入300 IU/ml IL-2及等量抗CD3/CD28 mAb包被磁珠,行混合淋巴细胞共培养,于培养结束前18 h加入3H-TdR,共培养7 d后检测各组细胞CPM增殖情况。5 μg/ml gD蛋白处理5例不同个体Treg后将其细胞浓度调整为5×106并接种于6孔培养板内,加入500 IU/ml IL-2共培养7 d,行RT-PCR(Realtime PCR)分析Treg Foxp3基因表达。    结果    HVEM在初始CD4+CD25+ Tregs表面呈中度表达,平均表达率为42%,且随着Treg的活化其表达HVEM上调;CD160在初始CD4+CD25- T细胞表面呈低度表达,平均表达率仅7%,但伴随其活化,表达CD160有所上调。采用不同浓度HSV1 gD蛋白处理Treg后2.5 μg/ml与5 μg/ml浓度组抑制CD4+CD25- T细胞增殖作用与空白对照组(276±35)相比明显下降,CPM值分别为(2 014±202)、(6 535±531),差异具有统计学意义(P<0.05),且具有浓度依赖性;RT-PCR分析发现Treg经5 μg/ml gD蛋白处理后Foxp3基因表达率下降,仅为空白对照组的48%,差异具有统计学意义(P<0.01)。    结论    机体接受免疫刺激后Treg通过上调HVEM表达与CD4+CD25- 效应性T细胞(effective T cell,Teff)表面上调表达的受体CD160交联从而上调Treg Foxp3基因的表达,最终介导了Treg对Teff活化、增殖的抑制作用,HVEM-CD160通路对Treg免疫调节作用具有重要意义。
Abstract:
Objective    To study the expression of herpes virus entry mediator (HVEM) and CD160 in different CD4+ T cell subsets under different function states and the influences of HVEM-CD160 pathway on CD4+CD25+ regulatory T cells (Tregs).     Methods    Flow cytometry was used to detect HVEM and CD160 expression in CD4+CD25+ Tregs and CD4+CD25- T cells from 3 different individuals at 0, 3, and 5 d after stimulation by 1 000 IU/ml human IL-2 and magnetic beads (with equal number to the cells) coated with monoclonal antibodies against CD3 and CD28. Tregs from 4 different individuals were treated with HSV1 gD protein (0, 1.25, 2.5, and 5 μg/ml separately), mixed with equivalent CD4+CD25- T cells and the magnetic beads and 300 IU/ml IL-2 in vitro, and cultured for 7 d. 3H-TdR was added to the culture liquid 18 h before stopping the culture, and CPM value was detected. Tregs from 5 different individuals were treated with 5 μg/ml gD proteins, adjusted to a concentration of 5×106, transferred to a 6-well plate, and cultured with 500 IU/ml IL-2 for 7 d. Treg Foxp3 gene expression was detected by real-time RT-PCR.     Results    HVEM expression was at a medium level on naive CD4+CD25+ Treg surfaces with an average expression rate of 42%, and was up-regulated along with Treg activation. CD160 expression was at a low level on naive CD4+CD25-  T cell surfaces with an average expression rate of 7%, and was up-regulated along with CD4+CD25- T cell activation. Tregs treated with 2.5 and 5 μg/ml HSV1 gD protein had significantly lowered and concentration-dependent inhibition effects on CD4+CD25- T cell proliferation (average CPM value 2 014±202 and 6 535±531) as compared with Tregs without treatment (average CPM value 276±35, P<0.05). RT-PCR results showed that Tregs treated with 5 μg/ml gD protein had a decreased Foxp3 gene expression rate, only 48% that of Tregs without treatment (P<0.01).     Conclusion    After immunostimulation, up-regulated HVEM in activated Tregs and up-regulated CD160 in activated CD4+CD25-  effector T cells (Teffs) interact to promote Treg Foxp3 gene expression, resulting in Treg’s inhibition effects on Teff activation and proliferation. HVEM-CD160 pathway is important for Treg’s inhibitory function.

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更新日期/Last Update: 2011-05-12