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Development of the new vaccine and a delivery vector by the recombination Morbilli virus.

利用重组麻疹病毒开发新疫苗和递送载体。

基本信息

批准号：
14360186
负责人：
KOHARA Kyoko
金额：
$ 10.82万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14360186/
关键词：
New Reverse Genetics method Morbilli virus vaccine Deliver vector

项目摘要

It aims at development of the safe recombination oral vaccine, half-oral vaccine and a new delivery vector by the new reverse genetics method which we established and which rearranges and makes development of a recombinant Morbilli virus possible in this research. This fiscal year mainly performed fundamental analysis of a virus vector using the rinderpest virus (RPV). First, it suggested that P protein was greatly concerned with the pathogenical change. when exchange the host using the RBOK and L strain of RPV. Moreover, to virus protein which governs pathogenicity was solved. Virus cloning established the highly pathogenic RPV-Lv strain and the weak pathogenic RPV-La strain out of the RPV-L strain. Consequently, among both, the amino acid substitution per site was found out in N, P, C, and L protein. When analyzed using seven sorts of viruses which rearranged the variation of N, P, and L genes of La strain with L strain, independently or simultaneous using the reverse genetics system … More , clinical condition was mitigated only for what changed N gene to La strain type. It became clear that N protein is important for pathogenesis of L strain. Moreover, in the B95a cell, although the vector which makes conditional expression system of M gene using a Cre/loxP system was produced for half-oral vaccine production, since the cell strain could nott be established. H gene mutated measles (MV) vector was tried to established, but a rescue was not succeeded, therefore, the new receptor binding unit which. is containing the penetration domain of the antibody recognition part gene (Fv) was induced in MV vector. They were obtained and analyzing whether an infection capacity has been changed. Furthermore, in order to analyze easily the delivery vector, the virus incorporating GFP and the luciferase gene was produced using the canine distemper virus (CDV) and MV vector. Among these, it succeeded in a MV-GFP vector visualizing the nerve cells which used the in vivo imaging equipment after inoculation in the mouse brain. Less

本研究旨在利用我们建立的新的反向遗传学方法，通过重组重组病毒，研制安全的重组口服疫苗、半口服疫苗和新的载体。本年度主要使用牛瘟病毒（RPV）进行病毒载体的基础分析。提示P蛋白与该病的发病密切相关。用RBOK和RPV L株交换宿主时，此外，还解决了病毒蛋白质的致病性问题。从RPV-L株系中克隆出高致病性RPV-Lv株系和弱致病性RPV-La株系。因此，在两者中，在N、P、C和L蛋白质中发现了每个位点的氨基酸替换。用7种病毒分别或同时用反向遗传学系统分析La株与L株N、P、L基因的变异， ...更多信息只有N基因改变为La株型才能缓解临床症状。由此可见，N蛋白在L株致病过程中起重要作用。此外，在B 95 a细胞中，虽然制备了用于半口服疫苗生产的使用Cre/loxP系统的M基因条件表达系统的载体，但由于不能建立细胞株。H基因突变的麻疹病毒（MV）载体尝试建立，但拯救失败，因此，新的受体结合单元，在MV载体中诱导了含有抗体识别部分基因（Fv）的穿透结构域。它们被获得并分析感染能力是否已经改变。此外，为了容易地分析递送载体，使用犬瘟热病毒（CDV）和MV载体制备掺入GFP和荧光素酶基因的病毒。其中，在接种于小鼠脑后使用体内成像设备的MV-GFP载体使神经细胞可视化是成功的。少