未经证实:HBV具有狭窄的宿主限制,人类和黑猩猩是唯一已知的自然宿主。在生物医学研究中常用的物种中,抗性的分子相关性,比如老鼠,目前还没有完全理解。人NTCP(hNTCP)在小鼠肝细胞中的表达使HBV进入,但随后共价闭合环状(cccDNA)不会在大多数鼠细胞中形成。尚不清楚cccDNA形成中的这种阻断是否是由于松弛环状DNA(rcDNA)对cccDNA的修复不足。
未经评估:这里,我们部署了体内和体外病毒学和生物化学方法,以研究鼠细胞是否含有一整套能够将HBVrcDNA转化为cccDNA的修复因子。
UNASSIGNED:我们证明,当没有蛋白质加合物的重组rcDNA直接引入细胞时,HBVcccDNA确实在鼠细胞培养或小鼠中形成。我们进一步表明,在DNA滞后链合成的核心成分的鼠直系同源物,在人类细胞中修复rcDNA到cccDNA所需的,可以支持HBV生命周期中的这一关键步骤。值得注意的是,重组HBVrcDNA底物,没有蛋白质加合物或含有中性抗生物素蛋白来模拟HBV聚合酶,在我们的研究中使用;尚不清楚HBV聚合酶去除过程是否在小鼠和人类细胞中相同。
未经批准:集体,我们的数据表明,HBV生命周期被阻断后进入和可能在小鼠细胞修复阶段之前,这产生了关键的见解,这将有助于构建具有近交易感性HBV感染的小鼠模型。
未经证实:乙型肝炎病毒(HBV)仅在自然界中感染人类和黑猩猩。小鼠模型通常用于建模疾病发病机理和临床前研究,以评估干预措施的有效性和安全性,然后在人类参与者中进行测试。然而,由于小鼠不易感染HBV,因此很难在小鼠模型中准确模拟人类感染(和测试潜在的治疗方法)。在这里,我们已经表明,小鼠能够执行HBV生命周期的关键步骤,收紧网络周围的可能原因HBV不能有效地感染和复制小鼠。
UNASSIGNED: HBV has a narrow host restriction, with humans and chimpanzees representing the only known natural hosts. The molecular correlates of resistance in species that are commonly used in biomedical research, such as mice, are currently incompletely understood. Expression of human NTCP (hNTCP) in mouse hepatocytes enables HBV entry, but subsequently covalently closed circular (cccDNA) does not form in most murine cells. It is unknown if this blockade in cccDNA formation is due to deficiency in repair of relaxed circular DNA (rcDNA) to cccDNA.
UNASSIGNED: Here, we deployed both in vivo and in vitro virological and biochemical approaches to investigate if murine cells contain a complete set of repair factors capable of converting HBV rcDNA to cccDNA.
UNASSIGNED: We demonstrate that HBV cccDNA does form in murine cell culture or in mice when recombinant rcDNA without a protein adduct is directly introduced into cells. We further show that the murine orthologues of core components in DNA lagging strand synthesis, required for the repair of rcDNA to cccDNA in human cells, can support this crucial step in the HBV life cycle. It is worth noting that recombinant HBV rcDNA substrates, either without a protein adduct or containing neutravidin to mimic HBV polymerase, were used in our study; it remains unclear if the HBV polymerase removal processes are the same in mouse and human cells.
UNASSIGNED: Collectively, our data suggest that the HBV life cycle is blocked post entry and likely before the repair stage in mouse cells, which yields critical insights that will aid in the construction of a mouse model with inbred susceptibility to HBV infection.
UNASSIGNED: Hepatitis B virus (HBV) is only known to infect humans and chimpanzees in nature. Mouse models are often used in modeling disease pathogenesis and preclinical research to assess the efficacy and safety of interventions before they are then tested in human participants. However, because mice are not susceptible to HBV infection it is difficult to accurately model human infection (and test potential treatments) in mouse models. Herein, we have shown that mice are able to perform a key step in the HBV life cycle, tightening the net around the possible reason why HBV can not efficiently infect and replicate in mice.