Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, USA.
Department of Microbiology and Center of Infectious Disease, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
Sci Rep. 2017 Nov 30;7(1):16616. doi: 10.1038/s41598-017-16882-5.
The discovery of sodium taurocholate cotransporting polypeptide (NTCP) as the hepatitis B virus (HBV) receptor enabled researchers to create hepatoma cell lines susceptible to HBV infection. Infection in current systems, however, is inefficient and virus fails to spread. Infection efficiency is enhanced by treating cells with polyethylene glycol 8000 (PEG) during infection. However, this alone does not promote virus spread. Here we show that maintaining PEG in culture medium increases the rate of infection by at least one order of magnitude, and, most importantly, promotes virus spread. To demonstrate the utility of this system, we show that two interferon-stimulated genes (ISGs), ISG20 and tetherin, restrict HBV spread in NTCP-expressing hepatoma cells. Thus, this protocol can be easily applied to existing cell culture systems to study the complete HBV life cycle, including virus spread.
牛磺胆酸钠共转运蛋白（NTCP）的发现使乙型肝炎病毒（HBV）受体得以鉴定，从而使研究人员能够创建对 HBV 感染敏感的肝癌细胞系。然而，目前的系统中感染效率低下，病毒无法传播。在感染过程中用聚乙二醇 8000（PEG）处理细胞可提高感染效率。但是，仅此并不能促进病毒传播。在这里，我们表明，在培养基中维持 PEG 可将感染率提高至少一个数量级，并且最重要的是，促进病毒传播。为了证明该系统的实用性，我们表明两种干扰素刺激基因（ISGs），ISG20 和 tetherin，可限制 NTCP 表达的肝癌细胞中的 HBV 传播。因此，该方案可以轻松应用于现有的细胞培养系统，以研究完整的 HBV 生命周期，包括病毒传播。
