Yao Yunqing, Huang Ailong, Tang Ni, Wang Bo, Zhang Dingfeng
Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2002 Feb;10(1):34-6.
By studying the possibility of obtaining expression of human hepatitis B virus (HBV) genes and production in normal liver cells from heterologous species like normal primary duck hepatocytes (PDH), to investigate the species-specificity of HBV infection and replication.
Two days after transfecting the complete HBV genome into PDH by electroporation (transfected group), HBsAg and HBeAg in the supernatants and lysates of PDH were measured by the IMX system. Meanwhile, replication of HBV in PDH was analyzed by Southern blotting and dot blotting procedures. PDH was electroporated as control.
HBsAg in the lysate of transfected group was 9.10 (P/N values, positive?2.1), HBeAg was 1.0 (negative?2.1), both were negative in the supernatants of transfected group. dot blotting revealed that transfected group was strongly positive, whereas the control group was negative. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (RC), covalently closed circular (CCC) and single-stranded (SS) HBV DNA replicative intermediates in the transfected group, and there was no integrated HBV DNA in the cellular genome. Control groups were negative.
Replication of HBV can occur in hepatocytes from nonmammalian species, which strongly supports the idea that replication of HBV has no critical species-specificity, and yet it depends on the endoenvironment of hepatocyte.
通过研究在正常原代鸭肝细胞（PDH）等异源物种的正常肝细胞中获得人乙型肝炎病毒（HBV）基因表达及产生病毒的可能性，探讨HBV感染和复制的种属特异性。
通过电穿孔将完整的HBV基因组转染到PDH中2天后（转染组），用IMX系统检测PDH上清液和裂解物中的HBsAg和HBeAg。同时，通过Southern印迹和斑点印迹法分析HBV在PDH中的复制情况。以电穿孔处理的PDH作为对照。
转染组裂解物中的HBsAg为9.10（P/N值，阳性＞2.1），HBeAg为1.0（阴性＜2.1），转染组上清液中的两者均为阴性。斑点印迹显示转染组呈强阳性，而对照组为阴性。细胞内总DNA的Southern印迹分析表明，转染组存在松弛环状（RC）、共价闭合环状（CCC）和单链（SS）HBV DNA复制中间体，细胞基因组中无整合的HBV DNA。对照组均为阴性。
HBV可在非哺乳动物物种的肝细胞中复制，这有力地支持了HBV复制无严格种属特异性但依赖于肝细胞内环境的观点。
