Hepatitis C virus infection in human liver tissue engrafted in mice with an infectious molecular clone
Hepatitis C virus infection in human liver tissue engrafted in mice with an infectious molecular clone
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DOI：
10.1111/j.1478-3231.2004.0909.x
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发表时间：
2004-06-01
影响因子：
6.7
通讯作者：
Hibi, T
中科院分区：
文献类型：
--
作者：
Maeda, N;Watanabe, M;Hibi, T
背景/目标：近年来，丙型肝炎病毒（HCV）分子克隆的研究取得了新的进展，使我们能够将一些可用的HCV分子克隆应用于实验研究。然而，这些研究仅限于黑猩猩模型或从树鼩“分离的肝细胞”。在这项研究中，我们将“人肝组织”移植到免疫缺陷小鼠中，并使用感染性分子克隆研究HCV感染。研究方法：将来自正常（非HCV感染）肝脏的人肝组织移植到非肥胖糖尿病/严重联合免疫缺陷（NOD/SCID）小鼠中。然后，我们用HCV感染患者的血清或感染性HCV分子克隆接种小鼠。采用巢式逆转录聚合酶链反应（PCR）、实时检测PCR和原位PCR检测HCV RNA。结果：在没有任何生长支持的情况下，正常人肝组织在NOD/SCID小鼠中存活，同时保持原始的活性肝结构。接种后第4周，小鼠血清中检测到HCV RNA。原位PCR和免疫组化清楚地表明，HCV感染的肝细胞的细胞质中的阳性信号，而移植的人肝组织没有表现出明显的形态学变化，感染的指示。结论：将人肝组织移植到NOD/SCID小鼠体内，感染HCV分子克隆，可为HCV感染提供一种逆转遗传策略。
Background/aims: Recent advances in molecular cloning of hepatitis C virus (HCV) have enabled us to apply some available HCV molecular clones to experimental studies. However, these investigations have been restricted to chimpanzee models or 'isolated hepatocytes' from tree shrews. In this study, we engrafted 'human liver tissue' into immunodeficient mice and investigated HCV infection using an infectious molecular clone. Methods: Human liver tissues from normal (non-HCV-infected) liver were transplanted into non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. We then inoculated the mice with sera from HCV-infected patients or an infectious HCV molecular clone. HCV RNA was assessed using nested reverse-transcription polymerase chain reaction (PCR), real-time detection PCR and in situ PCR. Results: Without any growth support, normal human liver tissues survived in NOD/SCID mice while maintaining the original viable hepatic architecture. HCV RNA was detected in the mice serum until the fourth week after the inoculation. In situ PCR and immunohistochemistry clearly demonstrated positive signals for HCV in the cytoplasm of infected hepatocytes, while the engrafted human liver tissues showed no apparent morphological changes indicative of infection. Conclusion: Engraftment of human liver tissues into NOD/SCID mice and infection with HCV molecular clones could offer a reverse genetic strategy for HCV infection.