Infection of human hepatocyte chimeric mouse with genetically engineered hepatitis B virus
Infection of human hepatocyte chimeric mouse with genetically engineered hepatitis B virus
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DOI：
10.1002/hep.20892
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发表时间：
2005-11-01
期刊：
影响因子：
13.5
通讯作者：
Yoshizato, K
中科院分区：
文献类型：
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作者：
Tsuge, M;Hiraga, N;Yoshizato, K
由于缺乏长期感染克隆HBV的小动物模型，对乙型肝炎病毒（HBV）突变体的研究一直受到阻碍。利用肝细胞被人类肝细胞高度替代的小鼠模型，我们进行了HBV的传播实验，这些肝细胞存活时间很长，具有成熟的人类肝细胞功能。接种含HBV的人血清和瞬时或稳定转染1.4基因组长度HBV DNA的HepG2细胞系产生的病毒。制备了基因修饰的e抗原阴性突变株，并接种到小鼠模型中。在接种hbv阳性人血清样本的小鼠中观察到高水平（约10（10）拷贝/mL）病毒血症。在人肝细胞替代指数持续高的小鼠中，病毒血症水平趋于高。在注射体外生成的HBV的小鼠中也观察到高水平和持久的病毒血症。病毒血症持续了22周，直到死亡或致死。传代实验表明，这些小鼠的血清中含有感染性HBV。基因工程乙型肝炎抗原阴性突变克隆也被证明具有传染性。拉米夫定有效地降低了这些感染小鼠的病毒血症水平。总之，该小鼠HBV感染模型为HBV病毒学研究和抗HBV药物评价提供了一个有用的工具。我们的研究结果表明，HBeAg对于活性病毒的产生和传播是必不可少的。
Studies of hepatitis B virus (HBV) mutants have been hampered by the lack of a small animal model with long-term infection of cloned HBV. Using a mouse model in which liver cells were highly replaced with human hepatocytes that survived over a long time with mature human hepatocyte function, we performed transmission experiments of HBV. Human serum containing HBV and the virus produced in HepG2 cell lines that transiently or stably transfected with 1.4 genome length HBV DNA were inoculated. Genetically modified e-antigen-negative mutant strain also was produced and inoculated into the mouse model. A high-level (approximate to 10(10) copies/mL) viremia was observed in mice inoculated with HBV-positive human serum samples. The level of viremia tended to be high in mice with a continuously high human hepatocyte replacement index. High levels and long-lasting viremia also were observed in mice injected with the in vitro generated HBV. The viremia continued up to 22 weeks until death or killing. Passage experiments showed that the serum of these mice contained infectious HBV. Genetically engineered hepatitis B e antigen-negative mutant clone also was shown to be infectious. Lamivudine effectively reduced the level of viremia in these infected mice. In conclusion, this mouse model of HBV infection is a useful tool for the study of HBV virology and evaluation of anti-HBV drugs. Our results indicate that HBeAg is dispensable for active viral production and transmission.