权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Research for relationship between infectiosity and amino acid sequences of hepatitis C virus from human inocula and transmitted chimpanzee sera

人接种物和传播黑猩猩血清丙型肝炎病毒传染性与氨基酸序列关系的研究

基本信息

批准号：
06670241
负责人：
SUGITANI Masahiko
金额：
$ 1.34万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670241/
关键词：
Hepatitis C virus gene Infectious Titer HCV RNA 核酸変異チンパンジー感染実験

项目摘要

We analyzed nucleic and amino acid sequences of the hypervariable region (HVR) of the hepatitis C virus (HCV) in the 4 kinds of sera, for the study of the next step of the titration experiments using chimpanzees as follows. One ml each of 10^<-5>,10^<-3> and 10^<-2> dilutions of human serum #4 were subsequently inoculated to chimpanzees nos.364,346, and 345, respectively, and one ml of 10^<-4>,10^<-5>, and 10^<-6> dilutions of human serum #6 were subsequently inoculated to chimpanzee nos.353,215, and 340, respectively. Only chimpanzee nos.353 and 215 were positive for HCV RNA in the sera, while both of inocula #4 and #6 contained high titer of HCV RNA (2x10^6 copies/ml, each, by semi-quantitative competitive assay). RNA extraction, transcription to cDNA,and amplification were done, 20 colonies were picked up, and nucleic and amino acid sequences were determined. Genetic variations were limited mostly to the HVR region in clones from #4 and #6. There were 6 and 4 clones from #4 and #6, respectively, and clones from #4 were different from those from #6 in amino acid sequences (table 1 & 2). On the other hand, clones of chimpanzee nos.353 and 215 were completely same in amino acid sequences. Homology of all clones from both chimpanzees was identical to the major type (6a-1) of human inoculum #6. The results of titration experiment suggested that HCV RNA titer in sera was not always correlate with in vivo infectivity, and the analytical study of amino acid sequences suggested that major group clone in the diluted inocula were mainly transmitted and replicated.

我们分析了4种血清中丙型肝炎病毒（HCV）高变区（HVR）的核酸和氨基酸序列，以研究下一步使用黑猩猩进行的滴定实验。随后将10 μ g<-5>、10 μ g<-3>和10 μ g<-2>稀释的人血清#4各1 ml分别接种于364、346和345号黑猩猩，随后将10 μ g、<-4>10 μ g<-5>和10 μ g<-6>稀释的人血清#6各1 ml分别接种于353、215和340号黑猩猩。只有353号和215号黑猩猩血清中的HCV RNA呈阳性，而接种物#4和#6均含有高滴度的HCV RNA（通过半定量竞争性测定，各为2 × 10^6拷贝/ml）。进行RNA提取、转录成cDNA和扩增，挑取20个菌落，测定核酸和氨基酸序列。遗传变异主要限于来自#4和#6的克隆中的HVR区域。分别有来自#4和#6的6个和4个克隆，并且来自#4的克隆在氨基酸序列上不同于来自#6的那些（表1和2）。另一方面，黑猩猩353号和215号克隆的氨基酸序列完全相同。来自两种黑猩猩的所有克隆的同源性与人接种物#6的主要类型（6a-1）相同。滴定实验结果表明血清中HCVRNA滴度与体内感染性并不总是相关的，氨基酸序列分析表明稀释接种物中的主群克隆主要是传播和复制的。