Novel reporter system to monitor early stages of the hepatitis B virus life cycle.
Novel reporter system to monitor early stages of the hepatitis B virus life cycle.
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DOI：
10.1111/cas.12799
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发表时间：
2015-11
期刊：
影响因子：
5.7
通讯作者：
Shimotohno K
中科院分区：
文献类型：
--
作者：
Nishitsuji H;Ujino S;Shimizu Y;Harada K;Zhang J;Sugiyama M;Mizokami M;Shimotohno K
表达NanoLuc（NL）的重组B肝炎病毒（HBV）（HBV/NL）是通过将含有携带NL基因的1.2倍HBV基因组的质粒与含有包装缺陷型1.2倍HBV基因组的质粒共转染到人肝癌细胞系HepG 2中产生的。我们发现，在感染HBV/NL的原代肝细胞或牛磺胆酸钠共转运多肽的人肝细胞衍生细胞系中，NL活性在感染后几天内呈线性增加，并且与细胞中的HBV RNA水平一致。用HBV抑制剂处理病毒感染的细胞以剂量依赖性方式降低NL活性。检测HBV/NL感染，监测NL活性，是高度敏感的，比使用常规方法来评估HBV感染的检测成本更低。此外，由于我们还研究了宿主因素，因此该系统不仅适用于研究HBV生命周期，还适用于探索调节HBV增殖的试剂。
A recombinant hepatitis B virus (HBV) expressing NanoLuc (NL) (HBV/NL) was produced by cotransfecting a plasmid containing a 1.2‐fold HBV genome carrying the NL gene with a plasmid bearing a packaging‐defective 1.2‐fold HBV genome into a human hepatoma cell line, HepG2. We found that NL activity in HBV/NL‐infected primary hepatocytes or sodium taurocholate cotransporting polypeptide‐transduced human hepatocyte‐derived cell lines increased linearly for several days after infection and was concordant with HBV RNA levels in the cells. Treatment of the virus‐infected cells with HBV inhibitors reduced NL activity in a dose‐dependent manner. Detection of HBV/NL infection, monitored by NL activity, was highly sensitive and less expensive than detection using the conventional method to evaluate HBV infection. In addition, because we also studied host factors, this system is applicable not only for studying the HBV life cycle, but also for exploring agent(s) that regulate HBV proliferation.