Lee I H, Kim C H, Ryu W S
Biotech Research Institute, LG Chemical Ltd/Research Park, Taejon, Korea.
J Med Virol. 1996 Oct;50(2):145-51. doi: 10.1002/(SICI)1096-9071(199610)50:2<145::AID-JMV7>3.0.CO;2-A.
Subviral particles of hepatitis B virus have been used to present foreign epitopes. We attempted to present the hydrophilic domains of E2 envelope protein of hepatitis C virus (HCV) as a fusion protein with hepatitis B virus surface antigen (HBsAg). The five hydrophilic domains of HCV E2 antigen were inserted into HBsAg such that the inserted hydrophilic domains were presented on the outer surface of HBV subviral particles. In addition, a fusion encoding the hypervariable region (HVR) of E2 antigen was also made. Cell lysate and culture medium were analyzed for the synthesis and secretion of the fusion proteins by immunoprecipitation with polyclonal anti-HBsAg antibody using recombinant vaccinia virus system. The results showed that the fusion proteins containing these six E2 domains were made in the cell, but only two out of six fusion proteins were secreted into culture medium. Further, cesium chloride density gradient analysis and electron microscopy revealed that these fusions were secreted into culture media as particles. It will be of interest to test immunogenicity of the HBsAg fusion particles containing the HCV E2 domains in animal model.
乙型肝炎病毒的亚病毒颗粒已被用于呈递外源表位。我们尝试将丙型肝炎病毒（HCV）E2包膜蛋白的亲水区作为与乙型肝炎病毒表面抗原（HBsAg）的融合蛋白来呈递。将HCV E2抗原的五个亲水区插入HBsAg中，使得插入的亲水区呈现在HBV亚病毒颗粒的外表面。此外，还构建了编码E2抗原高变区（HVR）的融合体。使用重组痘苗病毒系统，通过用多克隆抗-HBsAg抗体进行免疫沉淀，分析细胞裂解物和培养基中融合蛋白的合成与分泌情况。结果显示，含有这六个E2结构域的融合蛋白在细胞中产生，但六个融合蛋白中只有两个分泌到培养基中。此外，氯化铯密度梯度分析和电子显微镜显示，这些融合体以颗粒形式分泌到培养基中。在动物模型中测试含有HCV E2结构域的HBsAg融合颗粒的免疫原性将是很有意义的。
