Viral and host factors contributing to the neuro-virulence of rabies virus.

导致狂犬病病毒神经毒力的病毒和宿主因素。

基本信息

批准号：
08456150
负责人：
MINAMOTO Nobuyuki
金额：
$ 4.29万
依托单位：
Gifu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08456150/
关键词：
Rabies virus Glycoprotein gene Nucleotide substitution Revertant Deduced amino acid sequence Whole-cell patch clamp Na^+ channel Ultra structure 糖(G)遺伝子

项目摘要

In this study, to definite the molecular bases of neurovirulence in rabies virus, we have analyzed and characterized the pathogenic factors in the both virion and cells. The results run as follows :1) The RC-HL strain lacking the capacity to kill adult mice was passaged intracerebrally 22 times in suckling mice. Each passaged virus failed to regain lethal virulence for adult mice, although the viruses were increased to level of mouse pathogenicity.2) The revertant strain acquired partially virulence had each one amino acid substitution in G and M proteins. and one nucleotide substitution in non-coding region of 3' end in M gene when compared with full genome of the RC-HL strain. These changed positions of a nucleotide and two amino acids may necessary for mouse virulence.3) A comparative analysis of antigenic variants selected in the presence of virus-neutralizing monoclonal antibody (N-MAb) with cell-fusion inhibition activity, revealed that fusion activity corresponded to cystine at position 35 of the G protein.4) Any neutralization-resistant mutants by a N-MAb recognized an amino acid at position 333 of the (3 protein that related to virulence of rabies virus could not be selected from virulent virus, the Nishigahara strain, suggesting that arginine at the position is necessary for replication of virulent rabies virus.5) It was suggested that infection of mouse neuroblastoma NA cells with rabies virus causes reduction of functional expression of ion channels responsible for Na^+ current and delayed rectifier K^<1+> current, and provided evidence for possible involvement of the change in membrane properties in the pathogenesis of rabies disease.

在本研究中，为了确定狂犬病病毒神经毒力的分子基础，我们分析并表征了病毒颗粒和细胞中的致病因素。结果如下：1)将缺乏杀死成年小鼠能力的RC-HL菌株在乳鼠脑内传代22次。每种传代病毒均未能恢复对成年小鼠的致死毒力，尽管病毒已增加至小鼠致病性水平。2)获得部分毒力的回复株在G和M蛋白中各有一个氨基酸取代。与RC-HL株全基因组相比，M基因3'端非编码区有1个核苷酸取代。这些核苷酸和两个氨基酸位置的改变可能是小鼠毒力所必需的。3) 在具有细胞融合抑制活性的病毒中和单克隆抗体 (N-MAb) 存在下选择的抗原变体的比较分析表明，融合活性对应于 G 蛋白第 35 位的胱氨酸。4) N-MAb 的任何中和抗性突变体识别第 333 位的氨基酸（3 与狂犬病病毒毒力相关的蛋白质可以不选自强毒病毒西原株，表明该位置的精氨酸对于强毒狂犬病病毒的复制是必需的。5)有人认为，用狂犬病病毒感染小鼠神经母细胞瘤NA细胞会导致负责离子通道功能表达的减少Na^+电流和延迟整流K^<1+>电流，并为膜特性的变化可能参与狂犬病的发病机制提供了证据。