中文摘要：

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、5μg/ml HSV1gD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。5μg/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HSV1gD蛋白处理Treg后2.5μg/ml与5μg/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5μg/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（ef-fective Tcell,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。

英文摘要：

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,P0.05）.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（P0.01）.Conclusion After immunostimulation,up-regulated HVEM in activated Tregs and up-regulated CD160 in activated CD4＋CD25-effector T cells（Teffs