目的体外研究T细胞对感染人巨细胞病毒(human cytomegalovirus,HCMV)的绒毛外滋养细胞的免疫效应。方法从健康育龄妇女外周血中分离纯化获得T细胞,建立T细胞与感染HCMV的HPT-8共培养细胞模型,实时荧光定量PCR检测HPT-8内HCMVDNA负荷量。结果加入HCMV48h后,HPT-8胞质内出现大量HC.MVpp65抗原信号;T细胞具有下调HPT-8中HCMVDNA负荷量的作用(P〈0.05);T细胞与感染的HPT-8共培养3天后的细胞上清,也具有下调HPT-8中HCMVDNA负荷量的作用(P〈0.05);共培养3天后的T细胞则失去了下调HPT-8中HCMVDNA负荷量的作用(P〉0.05)。结论T细胞在母-胎界面HCMV感染初期具有一定的免疫防御作用,其机制可能与T细胞的直接溶解杀伤作用及其分泌的细胞因子相关;但T细胞的免疫功能很快受到抑制,其中的机制有待进一步研究。
Objective: To explore T cell-mediated restriction of human cytomagalovirns (HCMV) in trophoblastic ceils of external villus in vitro. Methods: T cells were isolated and purified from the peripheral blood of normal reproductive women. A T celI-HCMV infected HPT-8 co-culture system was established, and real-time quantitative PCR was used to de- tect the HCMV DNA in HPT-8. Results : A large number of HCMV pp65 antigen signals were seen in infected HPT-8 at the 48 time point. HCMV DNA load in infected HPT-8 was significantly decreased in T cells co-cultured group (P 〈 0.05). Cell-free co-culture supernatant was also diminished the HCMV DNA load (P 〈 0.05). However, when co-cultured T cells was added to fresh HCMV infected HFF-8, there was no significant difference in HCMV DNA load with HCMV infected group (P 〉 0.05 ). Conclusion: T cells have antiviral effect during the early stage of infected maternal-fetal interface, and the mechanism may be associated with both direct killing effect and secreting soluble factors. However, the immune function of T cells is rapidly inhibited, of which the underlying mechanisms remain unclear and need further study.