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目的检测牙龈卟啉单胞菌对人外周血中T淋巴细胞活化及凋亡的作用,并检测Fas/FasL在牙龈卟啉单胞菌(Porphyrom onas gingivalis,Pg)诱导的T淋巴细胞凋亡中的表达。方法选取10例全身及牙周组织健康受试者,分离外周血中T淋巴细胞,在有/无Pg情况下培养0~96 h,用荧光探针(Annexin V-FITC、PI、CD69)及特殊的单克隆抗体(Fas、FasL)进行标记,并进行流式细胞仪检测。结果 CD69+淋巴细胞+Pg组Annexin V+/PI-细胞百分数在各个时间点上都明显高于T淋巴细胞+Pg组(P<0.01)。Fas和FasL的表达量明显上调。用抗Fas单克隆抗体阻滞Fas-FasL相互作用导致T细胞凋亡的明显减少,百分比为(20.56±2.43)%,未加抗体的为(50.41±2.68)%。但残余的细胞凋亡活动与阴性对照相比仍高。结论 Pg能够诱导人外周血中T淋巴细胞活化,并且能够通过活化促进其凋亡,Pg诱导T淋巴细胞凋亡主要通过Fas-FasL途径,并具有时间依赖性。
Objective To detect the effect of Porphyromonas gingivalis on the activation and apoptosis of T lymphocytes in human peripheral blood and to detect the effect of Fas / FasL on the apoptosis of T lymphocytes induced by Porphyromonas gingivalis expression. Methods Ten healthy volunteers with whole body and periodontal tissues were selected and T lymphocytes were isolated from peripheral blood and cultured with and without Pg for 0-96 h. The expressions of PI and CD69 were detected by fluorescent probe (Annexin V-FITC) Specific monoclonal antibodies (Fas, FasL) were labeled and detected by flow cytometry. Results The percentage of Annexin V + / PI-cells in CD69 + lymphocytes + Pg group was significantly higher than that in T lymphocytes + Pg group at all time points (P <0.01). Fas and FasL expression was significantly increased. Blockade of Fas-FasL interaction with anti-Fas monoclonal antibody resulted in a significant reduction in T cell apoptosis, with a percentage of (20.56 ± 2.43)% and no antibody (50.41 ± 2.68)%. However, residual apoptotic activity was still high compared to the negative control. Conclusion Pg can induce activation of T lymphocytes in human peripheral blood and promote its apoptosis through activation. Pg-induced apoptosis of T lymphocytes mainly through the Fas-FasL pathway in a time-dependent manner.