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目的观察妊娠早期绒毛组织感染人巨细胞病毒(hCMV)后,即刻早期基因与晚期基因的表达及绒毛组织形态学变化。方法采用绒毛组织体外培养技术,建立体外hCMV感染妊娠早期绒毛模型;采用间接免疫荧光法和原位杂交法,检测不同hCMV浓度、不同感染时间,即刻早期蛋白(IEP)72-IEP86和晚期基因(LG)mRNA的表达;同时应用透射电镜观察妊娠早期绒毛组织的形态学变化。结果(1)以浓度为100半数致细胞病变滴度(TCID50)、200TCID50及300TCID50的hCMV感染绒毛组织后,细胞滋养细胞、合体滋养细胞及间质细胞均可见IEP72-IEP86表达。(2)100TCID50hCMV感染后,绒毛组织无LG mRNA表达;200TCID50及300TCID50hCMV感染第0～2天后,合体滋养细胞及间质细胞LG mRNA均呈阳性表达,细胞滋养细胞LG mRNA呈强阳性表达。(3)妊娠早期绒毛组织与经hCMV感染后的妊娠早期绒毛组织,共同于体外连续培养10d,妊娠早期绒毛组织保持了正常的形态学特征;不同浓度hCMV感染后的妊娠早期绒毛组织的形态均发生了不同程度的变化,合体滋养细胞表面微绒毛水肿、粗面内质网扩张,细胞滋养细胞多见,溶酶体增生及毛细血管腔扩张。结论(1)hCMV感染妊娠早期绒毛组织的早期,hCMV可在绒毛组织细胞中完整复制,IEP72-IEP86可长期存在于感染后的绒毛组织中。(2)hCMV感染妊娠早期绒毛组织,可引起绒毛组织细胞的超微结构变化。
Objective To observe the expression of early gene and late gene and the changes of villus histology after infection of human cytomegalovirus (hCMV) in early pregnancy. Methods The chorionic villus tissue culture technique was used to establish the early pregnancy villus model of hCMV infection in vitro. Indirect immunofluorescence and in situ hybridization were used to detect the expression of different hCMV concentrations, different infection time, immediate early protein (IEP) 72-IEP86 and late gene LG) mRNA expression was observed. Transmission electron microscopy was used to observe the morphological changes of villus during early pregnancy. Results (1) The expression of IEP72-IEP86 was found in cytotrophoblast, syncytiotrophoblast and mesenchymal cells after villus tissue was infected with TCCM50, 200TCID50 and 300TCID50 hCMV. (2) There was no LG mRNA expression in chorionic villi after infection with 100TCID50hCMV; LG mRNA was positive in syncytiotrophoblast and interstitial cells after 0 ~ 2 days of infection with 200TCID50 and 300TCID50hCMV; (3) In the first trimester of pregnancy, the chorionic villi in early pregnancy after hCMV infection were cultured continuously for 10 days in vitro. The normal early morphological features of chorionic villi maintained during the first trimester of pregnancy. Changes occurred in varying degrees, the surface of mixed trophoblastic microvilli edema, rough endoplasmic reticulum expansion, cytotrophoblast common, lysosomal proliferation and capillary lumen expansion. Conclusions (1) In the early stage of hCMV infection in early pregnancy villi, hCMV can be completely replicated in villus cells, and IEP72-IEP86 can persist in infected villus tissue. (2) hCMV infection of early pregnancy villi tissue, can cause villus tissue ultrastructure changes.