Rebound of hepatitis B virus replication in HepG2 cells after cessation of antiviral treatment
Rebound of hepatitis B virus replication in HepG2 cells after cessation of antiviral treatment
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DOI：
10.1128/jvi.76.16.8148-8160.2002
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发表时间：
2002-08-01
影响因子：
5.4
通讯作者：
Isom, HC
中科院分区：
文献类型：
--
作者：
Abdelhamed, AM;Kelley, CM;Isom, HC
使用拉米夫定 (3TC) 治疗患者会导致血清中可检测到的乙型肝炎病毒 (HBV) DNA 水平下降；然而，治疗终止后，病毒在血流中重新出现和肝脏炎症方面的复发率很高。尽管在动物肝炎病毒模型中也观察到在患者中观察到的反弹，但尚未在体外细胞培养系统中分析反弹。在本研究中，我们使用 HBV 重组杆状病毒/HepG2 系统来测量抗病毒药物介导的 HBV 复制丧失的时间过程，以及抗病毒治疗释放后 HBV 产生的时间过程和程度。由于系统的敏感性，可以测量分泌的病毒体、细胞内复制中间体和核非蛋白质结合的 HBV DNA，并单独分析各个 DNA 种类，例如单链 HBV DNA 与双链形式的比较，以及松弛环状 HBV DNA 与共价闭合环状 HBV DNA 的比较。我们首先确定，HBV重组杆状病毒/HepG2系统中的HBV复制可以持续至少35天，并具有30天的复制平台水平，使得研究抗病毒药物介导的HBV丢失以及停止药物治疗后反弹成为可能。所有 HBV DNA 种类在抗病毒治疗后均以时间依赖性方式下降，但每种 HBV DNA 种类下降的幅度不同，共价闭合环状 HBV DNA 对药物治疗的抵抗力最强。当药物治疗停止时，HBV DNA 物种重新出现，其模式重现了复制的起始，但时间进程不同。
Treatment of patients with lamivudine (3TC) results in loss of detectable levels of hepatitis B virus (HBV) DNA from serum; however, the relapse rate, with regard to both reappearance of virus in the bloodstream and hepatic inflammation, is high when therapy is terminated. Although the rebound observed in patients has also been seen in animal hepadnavirus models, rebound has not been analyzed in an in vitro cell culture system. In this study, we used the HBV recombinant baculovirus/HepG2 system to measure the time course of antiviral agent-mediated loss of HBV replication as well as the time course and magnitude of HBV production after release from antiviral treatment. Because of the sensitivity of the system, it was possible to measure secreted virions, intracellular replicative intermediates, and nuclear non-protein-bound HBV DNA and separately analyze individual species of DNA, such as single-stranded HBV DNA compared to the double-stranded form and relaxed circular compared to covalently closed circular HBV DNA. We first determined that HBV replication in the HBV recombinant baculovirus/HepG2 system could proceed for at least 35 days, with a 30-day plateau level of replication, making it possible to study antiviral agent-mediated loss of HBV followed by rebound after cessation of drug treatment. All HBV DNA species decreased in a time-dependent fashion following antiviral treatment, but the magnitude of decline differed for each HBV DNA species, with the covalently closed circular form of HBV DNA being the most resistant to drug therapy. When drug treatment ceased, HBV DNA species reappeared with a pattern that recapitulated the initiation of replication, but with a different time course.