Characterization of pseudotype VSV possessing HCV envelope proteins
Characterization of pseudotype VSV possessing HCV envelope proteins
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DOI：
10.1006/viro.2001.0971
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发表时间：
2001-08-01
期刊：
影响因子：
3.7
通讯作者：
Miyamura, T
中科院分区：
文献类型：
--
作者：
Matsuura, Y;Tani, H;Miyamura, T
丙型肝炎病毒（HCV）基因组编码两种包膜糖蛋白（E1和E2），它们被认为负责受体结合和膜融合，导致病毒穿透。为了研究HCV感染的重要细胞表面决定因素，我们使用了重组水泡性口炎病毒（VSV），其中糖蛋白基因被替换为报告基因编码的绿色荧光蛋白（GFP）和生产HCV-VSV假型具有嵌合的HCV E1或E2糖蛋白，无论是单独或一起。假型的感染性通过定量表达GFP报告基因的细胞的数量来确定。含有嵌合E1和E2蛋白的假型在HepG 2细胞上表现出比单独具有任一糖蛋白的病毒高10-20倍的感染性。这些结果表明，E1和E2包膜蛋白是HCV最大感染所必需的。假型病毒的感染性不能被抗VSV多克隆抗体中和，牛乳铁蛋白特异性地抑制假型病毒的感染。用链霉蛋白酶、肝素酶和肝素酶处理HepG 2细胞，但不用磷脂酶C和高碘酸钠处理，降低了感染性。因此，细胞表面蛋白和某些糖胺聚糖在HCV结合或进入易感细胞中起重要作用。具有嵌合HCV糖蛋白的假型VSV可能为进一步研究HCV细胞受体和开发丙型肝炎的抗病毒和治疗药物提供有效的工具。(C)北京：科学出版社.
The genome of hepatitis C virus (HCV) encodes two envelope glycoproteins (E1 and E2), which are thought to be responsible for receptor binding and membrane fusion resulting in virus penetration. To investigate cell surface determinants important for HCV infection, we used a recombinant vesicular stomatitis virus (VSV) in which the glycoprotein gene was replaced with a reporter gene encoding green fluorescent protein (GFP) and produced HCV-VSV pseudotypes possessing chimeric HCV E1 or E2 glycoproteins, either individually or together. The infectivity of the pseudotypes was determined by quantifying the number of cells expressing the GFP reporter gene. Pseudotypes that contained both of the chimeric E1 and E2 proteins exhibited 10-20 times higher infectivity on HepG2 cells than the viruses possessing either of the glycoproteins individually. These results indicated that both E1 and E2 envelope proteins are required for maximal infection by HCV. The infectivity of the pseudotype virus was not neutralized by anti-VSV polyclonal antibodies, Bovine lactoferrin specifically inhibited the infection of the pseudotype virus. Treatment of HepG2 cells with Pronase, heparinase, and heparitinase but not with phospholipase C and sodium periodate reduced the infectivity. Therefore, cell surface proteins and some glycosaminoglycans play an important role in binding or entry of HCV into susceptible cells, The pseudotype VSV possessing the chimeric HCV glycoproteins might offer an efficient tool for future research on cellular receptors for HCV and for the development of prophylactics and therapeutics for hepatitis C. (C) 2001 Academic Press.