伴放线聚集杆菌表面相关物质通过Fas-FasL诱导T淋巴细胞凋亡的体外研究

The study of the apoptosis of T lymphocytes by the fas and fas ligand pathway induced by surface-associated material from aggregatibacter actinomyceme-mcomitans

目的检测在伴放线聚集杆菌表面相关物质(SAM)诱导下,T细胞上Fas-Fas L的表达情况。方法选取健康受试者10名,抽取静脉血,提取外周血中单核细胞(PBMC),用SAM体外刺激PBMC,用单克隆抗体(Fas、Fas L)进行标记,上流式细胞仪进行检测。结果 Fas和Fas L的表达量明显升高。细胞培养24h,实验组Fas表达量为24.61±2.82,96h为66.67±3.18。在24h,Fas L的表达量为5.46±1.28,而96h为22.14±2.25。抗Fas单克隆抗体明显阻滞Fas-Fas L相互作用,最终导致凋亡T淋巴细胞数目减少至21.2±2.95,未加抗体的为49.96±4.07。但残余的细胞凋亡活动仍比阴性对照组高。结论 Fas-Fas L途径,在SAM诱导T淋巴细胞凋亡中起主要作用,并具有时间依赖性。

Objective Investigate the involvement of the Fas/Fas L-mediated apoptotic pathway in Surfaceassociated material（SAM）. from Aggregatibacter actinomycemem-comitans-induced T-cell apoptosis. Methods Ten healthy subjects participated in this study. Venous blood was drawn and peripheral blood mononuclear cells（PBMC）was isolated. Stimulate the T lymphocytes with SAM and the cells were labled by the special monoclonal antibodies and analyzed by flow cytometry. Results The stimulation of T cells with SAM leads to the up-regulation of Fas and Fas L.Fas： 24h-24.61 ±2.82, 96h-66.67 ±3.18; Fas L： 24h-5.46 ±1.28, 96h-22.14 ±2.25. The blocking of Fas-Fas L interaction with anti-Fas monoclonal antibody led to significantly reduce T-cell apoptosis： 21.2 ±2.95, 49.96 ±4.07 without the antibody, but the residual apoptosis activity higher than background remained. Conclusion SAM induces apoptosis of T lymphocytes by the Fas-Fas L pathway.Data obtained on the molecular mechanism of apoptosis and the effect is time-dependent.