伴放线放线杆菌诱导T淋巴细胞通过Fas-FasL途径的细胞凋亡的研究

The Study of the Apoptosis of T Lymphocytes by the Fas and Fas Ligand Pathway Induced by Actinobacillus Actinomycetemcomitans

目的 :检测与Aa诱导T淋巴细胞凋亡有关的Fas(CD95 )介导的细胞凋亡途径。方法 :选取 10名全身及牙周组织健康受试者 ,分离外周血单核细胞 (PBMC) ,Aa体外刺激PBMC ,用特殊的单克隆抗体 (Fas、FasL)标记 ,并进行流式细胞仪检测。结果 :Fas和FasL的表达量明显上调。实验组Fas:2 4h - 2 7.2 2 %± 4 .10 ,96h - 6 8.2 6 %± 3.14 ;FasL :2 4h 6 .18%± 1.37,96h - 2 2 .6 4 %± 2 .82。用抗Fas单克隆抗体阻滞Fas-FasL相互作用导致明显的T细胞凋亡减少 ,百分比为 2 2 .72 %± 3.5 4 ,未加抗体的为 5 1.4 7%± 3.75。 (P <0 .0 0 0 5 ,但残余的细胞凋亡活动比阴性对照仍高。结论 :Aa诱导T淋巴细胞主要通过Fas -FasL途径凋亡 ,在细胞凋亡的分子机制上得到了数据证明 。

Objective: Investigate the involvement of the Fas(CD95)-mediated apoptotic pathway in Actinobacillus actinocetemcomitans-induced T-cell apoptosis. Methods: Ten systemically and periodontally healthy subjects participated in this study. Venous blood was drawn and peripheral blood mononuclear cells(PBMC) was isolated. Stimulate the T lymphocytes with Aa and the cells were labled by the special monoclonal antibodies and analyzed by flow cytomety. Results:The stimulation of T cells with Aa leads to the up-regulation of Fas and FasL.Fas: 24h-27.22%±4.10,96h-68.26%±3.14; FasL:24h-6.18%±1.37, 96h-22.64%±2.82.The blocking of Fas-FasL interaction with anti-Fas monoclonal antibody led to significantly reduced T-cell apoptosis:22.72%±3.54 , 51.47%±3.75 without the antibody.(P<0.0005,but the residual apoptosis activity higher than background remained. Conclusions: Aa induces apoptosis of T lymphocytes by the Fas-FasL pathway .Data obtained on the molecular mechanism of apoptosis and the effect is time-dependent.