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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 To detect the expression of Fas-Fas L on T cells induced by SAM associated with actinomycetemcomitans. Methods Ten healthy volunteers were selected and venous blood samples were taken for extraction of peripheral blood mononuclear cells (PBMCs). PBMCs were stimulated with SAM in vitro and labeled with monoclonal antibodies (Fas and Fas L). The cytokines were detected by flow cytometry. Results The expression of Fas and Fas L increased significantly. The cell culture 24h, the experimental group of Fas expression was 24.61 ± 2.82, 96h 66.67 ± 3.18. At 24 h, the expression of Fas L was 5.46 ± 1.28, while it was 22.14 ± 2.25 at 96 h. Anti-Fas monoclonal antibody significantly blocked the Fas-Fas L interaction, eventually leading to a decrease in the number of apoptotic T lymphocytes to 21.2 ± 2.95 and no antibody to 49.96 ± 4.07. However, the remaining apoptotic activity was still higher than the negative control group. Conclusion The Fas-Fas L pathway plays a major role in SAM-induced apoptosis of T lymphocytes and is time-dependent.