An infectious molecular clone of a Japanese genotype 1b hepatitis C virus
An infectious molecular clone of a Japanese genotype 1b hepatitis C virus
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DOI：
10.1002/hep.510300137
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发表时间：
1999-07-01
期刊：
影响因子：
13.5
通讯作者：
Lemon, SM
中科院分区：
文献类型：
--
作者：
Beard, MR;Abell, G;Lemon, SM
我们描述了日本丙型肝炎病毒（HCV）基因型Ib株的感染性分子克隆。将HCV基因型Ib株的分子克隆序列与其它HCV序列进行比较，发现HCV基因型Ib株的开放阅读框（ORF）内有15个独特的非保守氨基酸替换，这些替换被修复到共有基因型1b残基上，并通过对黑猩猩的肝内接种来评估从修复的克隆转录的RNA的感染性。病毒RNA在接种后3周首次在这只黑猩猩的血清中检测到，并在接下来的14周内间歇性地存在。接种后13周出现强烈和持续的抗-HCV血清学应答，在重组免疫印迹试验（RIBA）中出现血清转换，在第3代酶联免疫吸附试验（ELISA）中出现较弱的一过性血清学应答，但在第1 - 5周未出现RIBA，这可能代表了对直接从肝内接种的RNA翻译的HCV抗原的记忆应答，因为该动物之前已经经历了2次通过肝内RNA接种拯救HCV的失败尝试。既没有生化也没有肝脏疾病的组织学证据。虽然这是在HCV未感染黑猩猩的预期结果范围内，但先前的免疫引发可能改变了该动物的感染。HCV-N克隆是HCV的第一个完全由基因型1b序列组成的感染性分子克隆，并且它含有与先前描述的基因型1a/1b嵌合体的ORF序列显著不同的ORF序列。
We describe an infectious molecular clone of a Japanese genotype Ib strain of hepatitis C virus (HCV-N), The molecularly cloned sequence of HCV-N was compared with alignments of other HCV sequences, leading to the identification of 15 unique, nonconservative amino acid substitutions within the HCV-N open reading frame (ORF), These were repaired to the consensus genotype 1b residue, and the infectivity of RNA transcribed from the repaired clone was assessed by intrahepatic inoculation of a chimpanzee. Viral RNA was first detected in the serum of this chimpanzee 3 weeks following inoculation, and was intermittently present over the next 14 weeks. A strong and persistent anti-HCV serological response developed 13 weeks following inoculation, with seroconversion in the recombinant immunoblot assay (RIBA), A weaker, transient serological response, characterized by seroconversion in a third-generation enzyme-linked immunosorbent assay (ELISA) but not RIBA, occurred between weeks 1 and 5, This may have represented an anamnestic response to HCV antigens translated directly from the intrahepatically inoculated RNA, because the animal previously had undergone 2 unsuccessful attempts at rescue of HCV by intrahepatic RNA inoculation. There was neither biochemical nor histological evidence of liver disease. Although this is within the range of expected outcomes in an HCV-naive chimpanzee, prior immunologic priming may have modified the infection in this animal. The HCV-N clone is the first infectious molecular clone of HCV that is comprised entirely of genotype 1b sequence, and it contains an ORF sequence that is significantly divergent from that of a previously described genotype 1a/1b chimera.