Chai Ning, Chang Ho Eun, Nicolas Emmanuelle, Gudima Severin, Chang Jinhong, Taylor John
Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111-2497, USA.
J Virol. 2007 Oct;81(20):10897-904. doi: 10.1128/JVI.00959-07. Epub 2007 Aug 1.
This study demonstrates that the envelope proteins of hepatitis B virus (HBV) could be incorporated into the lipid membrane of lentivirus pseudotype particles. The assembly procedure was initiated by the transfection of 293T cells with three plasmids: (i) a human immunodeficiency virus (HIV) packaging construct, (ii) a transfer plasmid expressing a reporter gene, and (iii) a plasmid expressing large (L), middle (M), and small (S) HBV envelope proteins. After 2 days, hepatitis B surface antigen and the antigenic forms of L, M, and S were detected at the cell surface by flow cytometry. Also, virus particles that were able to infect cultured primary human hepatocytes (PHH) were released. Under optimal conditions, 50% of PHH could be infected. In addition, the susceptibility of PHH and the resistance of other cell types to the pseudotype particles were similar to those observed for HBV and hepatitis delta virus (HDV), which shares the same L, M, and S. Furthermore, the infection of PHH by the pseudotype was sensitive to known inhibitors of HBV and HDV entry. These findings of specific and efficient infection of hepatocytes could be applicable to liver-specific gene therapy and may help clarify the attachment and entry mechanism used by HBV and HDV.
本研究表明，乙型肝炎病毒（HBV）的包膜蛋白可整合到慢病毒假型颗粒的脂质膜中。组装过程通过用三种质粒转染293T细胞启动：（i）人免疫缺陷病毒（HIV）包装构建体，（ii）表达报告基因的转移质粒，以及（iii）表达HBV大（L）、中（M）和小（S）包膜蛋白的质粒。2天后，通过流式细胞术在细胞表面检测到乙型肝炎表面抗原以及L、M和S的抗原形式。此外，还释放出能够感染培养的原代人肝细胞（PHH）的病毒颗粒。在最佳条件下，50%的PHH可被感染。此外，PHH的易感性和其他细胞类型对假型颗粒的抗性与乙型肝炎病毒（HBV）和丁型肝炎病毒（HDV）观察到的情况相似，HDV与HBV共享相同的L、M和S。此外，假型对PHH的感染对已知的HBV和HDV进入抑制剂敏感。这些关于肝细胞特异性和有效感染的发现可能适用于肝脏特异性基因治疗，并可能有助于阐明HBV和HDV使用的附着和进入机制。
