滋养细胞系（HPT-8）的建立和鉴定以及HBV在滋养细胞中分布的胶体金探针示踪研究
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摘要
乙型病毒性肝炎(乙肝)是世界流行性疾病，严重地危害着人们的生活和健康。在乙型肝炎病毒(HBV)的传播途径中，垂直传播是十分重要的一种途径，即HBV可通过胎盘引起胎儿感染，又称作HBV宫内传播。目前，乙肝表面抗原(HBsAg)携带孕妇的胎儿宫内感染率为5％～15％。孕期经宫内传播途径造成的胎儿HBV感染，用乙肝疫苗和高效价免疫球蛋白不能阻断，宫内感染是乙型肝炎免疫失败的首要原因。自1992年以来，我室对HBV宫内传播机制进行了宏观和微观两方面的系统研究，提出血源性途径和细胞源性途径并存的假说。许多研究应用免疫病理和分子生物学技术证实胎盘各层细胞可以感染HBV。胎盘滋养层细胞作为胎盘屏障的第一层细胞，直接与母亲血液接触，以往的组织病理学研究也已经得到滋养层细胞感染的证据，但其确切机理国内尚未见报道。因此对HBV在滋养细胞内活动的研究，成为控制乙肝宫内传播的关键环节之一。本研究采用胶体金标记结合透射电镜技术是一种高分辨率免疫定位需鉴定的抗原于不同亚细胞部位的有价值的技术，为细胞源性途径假说的提供支持。
目的 建立传代稳定的、生物学特征与体内细胞接近的滋养细胞株(HPT-8)；通过胶体金探针标记技术提供HBV进入HPT-8的依据，并建立能分泌HBsAg和HBeAg的转染HPT克隆株。
方法
①取妊娠6～9wk、发育正常、新鲜流产胎盘组织，挑取绒毛，用胰酶和胶原酶Ⅰ混合消化法消化人绒毛组织，用胰蛋白酶消化传代体外培
Hepatitis B is still a major epidemic worldwide and threatened people's life and health heavily. Vertical transmission is a very important route of HBV transmission. In this route, HBV could infect fetus through placentae, which is called one of intrauterine transmission. The rate of HBV intrauterine transmission is 5%~15% in babies of the hepatitis B surface antigen positive mothers. The acquired HBV infection of fetus in gestation can not be blocked by hepatitis B vaccine and hepatitis B immunoglubin. The intrauterine transmission is major reason for the failure of hepatitis B immunity. Since 1992, our department have performed macroscopic and microcosmic studies in this field and suggested that HBV intrauterine transmission might occur either through placental tears with transfusion of maternall blood into fetal circulation named "hematogenous transmission" or progressive infection of every placental layer until the virus got to fetus, which was named "transplacental cellular traffic". Many studies by immunophathologic and molecular biologic technology confirmed that HBV could infect the cells of placenta. Trophoblast cells, as the outermost covering of placental barrier, are directly bathed in the maternal blood. Some studies has been showed the evidences of HBV infection in the cells by morphologyical and pathological methods in sections of placental tissue. However, the mechanism of entry of HBV into trophoblast cells has not been reported yet. So the studies on the trace of HBV in the trophoblast cells have been one of the keys in controlling
目的 建立传代稳定的、生物学特征与体内细胞接近的滋养细胞株(HPT-8)；通过胶体金探针标记技术提供HBV进入HPT-8的依据，并建立能分泌HBsAg和HBeAg的转染HPT克隆株。
方法
①取妊娠6～9wk、发育正常、新鲜流产胎盘组织，挑取绒毛，用胰酶和胶原酶Ⅰ混合消化法消化人绒毛组织，用胰蛋白酶消化传代体外培
Hepatitis B is still a major epidemic worldwide and threatened people's life and health heavily. Vertical transmission is a very important route of HBV transmission. In this route, HBV could infect fetus through placentae, which is called one of intrauterine transmission. The rate of HBV intrauterine transmission is 5%~15% in babies of the hepatitis B surface antigen positive mothers. The acquired HBV infection of fetus in gestation can not be blocked by hepatitis B vaccine and hepatitis B immunoglubin. The intrauterine transmission is major reason for the failure of hepatitis B immunity. Since 1992, our department have performed macroscopic and microcosmic studies in this field and suggested that HBV intrauterine transmission might occur either through placental tears with transfusion of maternall blood into fetal circulation named "hematogenous transmission" or progressive infection of every placental layer until the virus got to fetus, which was named "transplacental cellular traffic". Many studies by immunophathologic and molecular biologic technology confirmed that HBV could infect the cells of placenta. Trophoblast cells, as the outermost covering of placental barrier, are directly bathed in the maternal blood. Some studies has been showed the evidences of HBV infection in the cells by morphologyical and pathological methods in sections of placental tissue. However, the mechanism of entry of HBV into trophoblast cells has not been reported yet. So the studies on the trace of HBV in the trophoblast cells have been one of the keys in controlling
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