INSERM, U1052, Cancer Research Center of Lyon (CRCL), Université de Lyon (UCBL1), CNRS UMR_5286, Centre Léon Bérard, Lyon, France.
Sanofi R&D, Infectious Disease Therapeutic Area, Marcy l'Etoile, France.
Antiviral Res. 2017 Sep;145:14-19. doi: 10.1016/j.antiviral.2017.07.006. Epub 2017 Jul 11.
Hepatitis B Virus (HBV) persists in infected hepatocytes as an episomal covalently-closed-circular DNA mini-chromosome, called cccDNA. As the main nuclear transcription template, HBV cccDNA is a key replication intermediate in the viral life cycle. Little is known about the mechanisms involved in its formation, maintenance and fate under antiviral therapies. This is mainly due to the lack of small immune-competent animal models able to recapitulate the entire HBV replication cycle, including formation of HBV cccDNA. Here we report that HBV cccDNA can be detected by Southern blot analyses in the liver of C57BL6 mice transduced with AAV-HBV. HBV cccDNA persists in the liver of these animals together with the AAV-HBV episome. We also set up a PCR strategy to distinguish the HBV cccDNA from the AAV-HBV episome. These suggest that the AAV-HBV/mouse model might be relevant to test drugs targeting HBV cccDNA regulation and persistence.
乙型肝炎病毒 (HBV) 以共价闭合环状 DNA 超螺旋（cccDNA）的形式存在于受感染的肝细胞中，作为一种游离的环状 DNA。作为主要的核转录模板，HBV cccDNA 是病毒生命周期中的关键复制中间体。然而，关于其在抗病毒治疗下的形成、维持和命运的机制知之甚少。这主要是由于缺乏能够重现整个 HBV 复制周期的小型免疫功能健全的动物模型，包括 HBV cccDNA 的形成。在这里，我们报告说，在接受 AAV-HBV 转导的 C57BL6 小鼠肝脏中，可以通过 Southern blot 分析检测到 HBV cccDNA。HBV cccDNA 与 AAV-HBV episome 一起在这些动物的肝脏中持续存在。我们还建立了一种 PCR 策略来区分 HBV cccDNA 和 AAV-HBV episome。这些结果表明，AAV-HBV/小鼠模型可能与测试针对 HBV cccDNA 调节和持久性的药物有关。
