腺病毒携带人乙型肝炎病毒表面前S_2抗体基因转染大鼠移植肝预防乙型肝炎病毒再感染  

Adenoviral vector encoding the whole humanized antibody gene of preS_2 antigen inhibiting the recurrence of hepatitis B virus after liver transplantation

作　　者：郭明高[1] 杨甲梅[1] 单云峰[1] 张辉[1] 李根丛[1] 崔贞福[1] 钱其军[1] 吴孟超[1] 

机构地区：[1]第二军医大学东方肝胆外科医院,上海200438

出　　处：《中华器官移植杂志》2005年第6期370-372,共3页Chinese Journal of Organ Transplantation

基　　金：上海市科技发展基金资助项目(004119014)

摘　　要：目的探讨腺病毒携带人乙型肝炎病毒表面前S2抗体基因预防乙型肝炎病毒(HBV)再感染移植肝的作用。方法将人乙型肝炎病毒表面前S2抗原人源化抗体基因(Ab-H-HBV-S2)克隆入5型腺病毒穿梭质粒载体,在293细胞中重组,产生重组型腺病毒。用重组腺病毒感染冷保存时的大鼠供肝,获得人乙型肝炎病毒表面前S2抗体(简称为:前S2抗体)的表达。观察前S2抗体在体外保护人原代肝细胞不受HBV感染的效果。结果用50%组织培养感染剂量法(TCID50)测定重组腺病毒滴度为5.1×1010PFU/ml。受鼠肝移植后3d时,血清中前S2抗体的表达量为(16.7±10.5)mg/L,7d时的表达量为(30.9±13.6)mg/L。当受鼠血清中前S2抗体浓度≥0.5mg/L时,与HBV和人原代肝细胞共同培养后,检测上清液中乙型肝炎病毒表面抗原(HBsAg)为阴性。结论全长Ab-H-HBV-S2基因克隆入腺病毒载体,感染大鼠供肝,获得人源化抗体的表达;表达的前S2抗体能保护人原代肝细胞不受HBV感染。Objective To investigate the adenoviral vector encoding the whole humanized antibody gene of pres_2 antigen for preventing the liver graft from infection by the hepatitis B virus. Methods The whole humanized antibody gene to pres_2 antigen was cloned into type 5 adenoviral shuttle plasmid pDC315. The corresponding recombinant virus was obtained by homologous recombination in 293 packaging cells. The virus containing the whole humanized antibody gene of preS_2 antigen was transfected into the rat liver graft during cold preservation. The effect of the recipient serum containing preS_2 antibody protecting human hepatocytes from hepatitis B virus infection was observed. Results The viral titer determined by TCID50 analysis was 5.1 ×10 10 PFU/ml. The concentration of preS_2 antibody was ( 16.7 ± 10.5 ) mg/L on the day 3 and ( 30.9 ± 13.6 ) mg/L on the day 7. The serum containing preS_2 antibody could protect human hepatocytes from hepatitis B virus infection in vitro when the concentration of the preS_2 antibody was more than 0.5 μg/ml. Conclusions The adenoviral vector encoding the whole humanized antibody gene of preS_2 protein were constructed successfully. The whole humanized antibody was expressed. The preS_2 antibody in recipient serum protecting human hepatocytes from hepatitis B virus infection was observed, which might be a new method to prevent hepatitis B re-infection after liver transplantation.

关 键 词：前S2抗体 人乙型肝炎 乙型肝炎病毒再感染 腺病毒携带 移植肝 大鼠 基因转染 乙型肝炎病毒(HBV) 乙型肝炎病毒表面抗原 人原代肝细胞 HBV感染 前S2抗原 293细胞 腺病毒感染 腺病毒载体 人源化抗体 基因预防 质粒载体 抗体基因 

分 类 号：R657.3[医药卫生—外科学]