Molecular biological studies on the neurovirulence of murine coronavirus

鼠冠状病毒神经毒力的分子生物学研究

基本信息

批准号：
02660322
负责人：
TAGUCHI Fumihiro
金额：
$ 1.28万
依托单位：
National Institute of Neuroscience, NCNP
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

A highly neurotropic murine coronavirus JHM strain has been studied as a model of viral agent to cause acute and subacute encephalomyelitis with demyelination. As a viral factor involving in the neurovirulence. the spike(S)protein comprising the projections on the virion surface is considered to be deeply involved. To study the mechanisms by which JHM strain exhibits the neurovirulence, we have done the molecular biological analysis of S protein of variant JHM viruses showing different neurovirulence. We first of all have isolated a highly neurovirulent variant, cl-2, for rats. This variant has been shown to have a larger S protein as compared with other variants with avirulence. We have isolated the cDNA from mRNA3 encoding the larger S protein and sequence analysis has identified that the larger S protein has 141 amino acid clusters which is missing in a small S protein of avirulent viruses. The 141 amino acid cluster may be implicated with high neurovirulence, although the precise role of the amino acids is yet unclear. We further studied on the larger S protein by expressing in mammalian cells with recombinant vaceinia viruses. The S protein expressed showed no difference in molecular size, immunogenicity or antigenicity as compared with the authentic S protein. Furthermore, the expressed S protein exhibited the fusionability in various cultured cells. These results suggest that the S protein expressed by recombinant vaceinia virus is appropriate for the study of biological activities of the S protein. Experiments are now in progress to see the neurovirulence of JHMV using the recombinant vaccinia viruses with large S protein cDNA.

已经研究了一种高度神经性鼠冠状病毒JHM菌株作为病毒剂的模型，以引起伴有脱髓鞘的急性和亚急性脑脊髓炎。作为涉及神经动力学的病毒因子。尖峰（S）蛋白包含在病毒体表面上的投影被认为是深入参与的。为了研究JHM菌株表现出神经动力电力的机制，我们对变体JHM病毒的S蛋白进行了分子生物学分析，显示出不同的神经动力学。首先，对于大鼠，我们首先隔离了高度神经性肺部的变体Cl-2。与其他具有活力的变体相比，该变体已被证明具有较大的S蛋白。我们已经从编码较大S蛋白的mRNA3中分离了cDNA，序列分析已经确定较大的S蛋白具有141个氨基酸簇，这在无毒病毒的小S蛋白中缺少。尽管氨基酸的确切作用尚不清楚，但141个氨基酸簇可能与高神经动力学有关。我们通过在哺乳动物细胞中用重组叶乙乙酰病毒表达较大的S蛋白。与正宗S蛋白相比，表达的S蛋白在分子大小，免疫原性或抗原性上没有差异。此外，表达的S蛋白在各种培养细胞中表现出融合性。这些结果表明，重组蔬菜病毒表达的S蛋白适用于研究S蛋白的生物学活性。现在正在进行实验，以观察使用具有大S蛋白cDNA的重组离子病毒的JHMV的神经电动性。