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

本研究旨在利用我们建立的新的反向遗传学方法，研制安全的重组口服疫苗、半口服疫苗和新的递送载体，使重组Morbilli病毒的研制成为可能。本财年主要使用牛瘟病毒(RPV)对一种病毒媒介进行了基本分析。首先，提示P蛋白与病原学变化密切相关。用RPV的RBOK株和L株交换宿主时。此外，还解决了控制致病性的病毒蛋白。病毒克隆建立了高致病性RPVLv株和弱致病力RPVLv株，而弱致病性RPVLv株和弱致病性RPVLv株是从RPV1株的L株中分离出来的。结果发现，在N、P、C和L蛋白中存在氨基酸的每一位点替代。利用反向遗传系统…对7种La株和L株N、P、L基因重排的病毒进行了单独或同时分析此外，只有将N基因改变为La株型的患者的临床症状才得到缓解。结果表明，N蛋白在L毒株的致病过程中起重要作用。此外，在B95a细胞中，虽然使用Cre/loxP系统建立M基因条件表达系统的载体是用于半口服疫苗生产的，但由于细胞株不能建立。试图建立H基因突变的麻疹(MV)载体，但营救没有成功，因此，新的受体结合单位。在MV载体中诱导含有抗体识别部分基因(FV)穿透结构域的IS。他们被获得并分析感染能力是否发生了变化。此外，为了便于分析，用犬瘟热病毒(CDV)和MV载体制备了携带GFP和荧光素酶基因的病毒。其中，成功地将MV-GFP载体接种到小鼠脑内后，利用体内成像设备对神经细胞进行了可视化。较少

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