Production of infectious genotype 1a hepatitis C virus (Hutchinson strain) in cultured human hepatoma cells
Production of infectious genotype 1a hepatitis C virus (Hutchinson strain) in cultured human hepatoma cells
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DOI：
10.1073/pnas.0510727103
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发表时间：
2006-02-14
影响因子：
11.1
通讯作者：
Lemon, SM
中科院分区：
文献类型：
--
作者：
Yi, M;Villanueva, RA;Lemon, SM
丙型肝炎病毒(丙型肝炎病毒)感染的特点是频繁的病毒持久性、慢性肝病和异常的病毒遗传多样性。虽然自从发现丙型肝炎病毒以来，人们已经了解了很多，但由于缺乏支持其复制的许可细胞培养系统，进展缓慢。最近，2a型病毒已经实现了生产性感染，但肝硬变和肝癌通常与1型丙型肝炎病毒有关，丙型肝炎病毒更普遍，对干扰素治疗相对耐药。我们描述了在用来自原型病毒(H77-S)的合成核糖核酸转染的细胞中产生具有感染性的基因型1丙型肝炎病毒。病毒蛋白在H77-S细胞中的积累较慢，但分泌到上清液中的H77-S RNA明显多于2a型(JFH-1)。分泌的核糖核酸多为非感染性，密度为1.04~1.07gm/cm(3)，呈等密度梯度条带，而感染性与密度为1.13~1.14gm/cm(3)的H77-S颗粒有关。H77-S颗粒的特异性感染力(每病灶单位5.4×10(4)个RNA拷贝数)明显低于JFH-1病毒0.4×10(2)个/病灶单位单位RNA拷贝数(P<0.05)。CD81抗体可阻断两种病毒的感染。1a型感染者血清对H77-S病毒有中和作用，但对2a型病毒活性较弱，提示不同的血清型代表不同的血清型。这种L型病毒感染培养细胞的能力将极大地有利于抗病毒和疫苗发现计划。
Infections with hepatitis C virus (HCV) are marked by frequent viral persistence, chronic liver disease, and extraordinary viral genetic diversity. Although much has been learned about HCV since its discovery, progress has been slowed by a lack of permissive cell culture systems supporting its replication. Productive infections have been achieved recently with genotype 2a virus, but cirrhosis and liver cancer are typically associated with genotype 1 HCV, which is more prevalent and relatively resistant to IFN therapy. We describe production of infectious genotype la HCV in cells transfected with synthetic RNA derived from a prototype virus (H77-S). Viral proteins accumulated more slowly in H77-S transfected cells than in cells transfected with genotype 2a (JFH-1) RNA, but substantially more H77-S RNA was secreted into supernatant fluids. Most secreted RNA was noninfectious, banding in isopycnic gradients at a density of 1.04-1.07 gm/cm(3), but infectivity was associated with H77-S particles possessing a density of 1.13-1.14 gm/cm(3). The specific infectivity of H77-S particles (5.4 x 10(4) RNA copies per focus-forming unit) was significantly lower than JFH-1 virus 0.4 x 10(2) RNA copies per focus-forming unit). Infection with either virus was blocked by CD81 antibody. Sera from genotype la-infected individuals neutralized H77-S virus, but had little activity against genotype 2a virus, suggesting that these genotypes represent different serotypes. The ability of this genotype la virus to infect cultured cells will substantially benefit antiviral and vaccine discovery programs.