Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras
Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras
复制标题
DOI：
10.1073/pnas.0504877103
复制
发表时间：
2006-05-09
影响因子：
11.1
通讯作者：
Bartenschlager, Ralf
中科院分区：
文献类型：
--
作者：
Pietschmann, Thomas;Kaul, Artur;Bartenschlager, Ralf
由丙型肝炎病毒（HCV）感染引起的慢性肝病是一个重要的全球健康问题，目前影响着1.7亿人。HCV研究和药物开发的一个主要障碍是缺乏支持病毒生产的培养系统。最近，通过使用基于JFH 1的全长基因组克服了这一障碍，该基因组允许在体外和体内产生感染性病毒。虽然这种改进是重要的，但由于JFH 1分离株和由J6 CF和JFH 1衍生序列组成的单一嵌合体的限制，不同HCV毒株之间的广泛基础的比较研究是不可能的。因此，在这项研究中，我们创建了一系列进一步的嵌合基因组，允许产生感染性基因型（GT）1a，1b，2a和3a颗粒。除了GT 3a/JFH 1嵌合体，当基因组片段通过位于NS 2的第一个跨膜结构域之后的位点融合时，获得了有效的病毒生产。最有效的构建体是GT 2a/2a嵌合体，其由通过该连接点连接的J6 CF-和JFH 1-衍生序列组成。这种杂交体，命名为Jc 1，产生的感染滴度比亲本分离株和所有其他嵌合体高100至1,000倍，表明结构蛋白内的决定因素控制病毒组装和释放的动力学和效率。最后，我们描述了E1特异性抗血清能够中和所有HCV嵌合体的感染性。
Chronic liver disease caused by infection with hepatitis C virus (HCV) is an important global health problem that currently affects 170 million people. A major impediment in HCV research and drug development has been the lack of culture systems supporting virus production. This obstacle was recently overcome by using JFH1-based full-length genomes that allow production of viruses infectious both in vitro and in vivo. Although this improvement was important, because of the restriction to the JFH1 isolate and a single chimera consisting of J6CF and JFH1-derived sequences, broadly based comparative studies between different HCV strains were not possible. Therefore, in this study we created a series of further chimeric genomes allowing production of infectious genotype (GT) 1a, 1b, 2a, and 3a particles. With the exception of the GT3a/JFH1 chimera, efficient virus production was obtained when the genome fragments were fused via a site located right after the first transmembrane domain of NS2. The most efficient construct is a GT2a/2a chimera consisting of J6CF- and JFH1-derived sequences connected via this junction. This hybrid, designated Jc1, yielded infectious titers 100-to 1,000-fold higher than the parental isolate and all other chimeras, suggesting that determinants within the structural proteins govern kinetic and efficiency of virus assembly and release. Finally, we describe an E1-specific antiserum capable of neutralizing infectivity of all HCV chimeras.