Analysis of the molecular mechanisms of host range determination of nucleopolyhedovirus

核多角体病毒宿主范围确定的分子机制分析

基本信息

批准号：
08456035
负责人：
KOBAYASHI Michihiro
金额：
$ 4.86万
依托单位：
Nagoya 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 1997
项目状态：
已结题

项目摘要

Autographa californica (Ac), Bombyx mori (Bm), Hyphantria cunea (Hc) and Spodoptera exigua (Se) nucleopolyhedroviruses (NPVs) were each inoculated to SF21cells, BmN-4 cells, SpIm cells and Se301 cells, and cytopathic effect (CPE) manifestation, viral DNA production, badded virion yield into the culture medium, viral structural protein production, polyhedrin (the matrix protein of polyhedra) production, and polyhedron formation in the inoculated cells were comparatively examined. The results showed that AcNPV were able to multiply and yield budded virions in all of four cell lines examined, but production of polyhedrin and polyhedra occurred only SF21cells, SpIm cells and Se301 cells, and not in BmN-4 cells.BmNPV multiplied to a high titre in BmN-4 cells and to a low titre in SpIm cells in which only a few number of cells manifested CPE.No clear CPE was observed in SF21 and Se301 cells inoculated with BmNPV.HcNPV multiplied unequivocally and produced polyhedra in SpIm and Se301 cells. In SF21 cells, HcNPV yielded budded virions only to a low titre and produced neither polyhedrin nor polyhedra. In HcNPV-inoculated BmN-4 cells, significant amount of viral DNA accumulated, but accumulation of viral structural protein and polyhedrin and budded virion yield were never detected. SeNPV multiplied clearly only in Se301 cells, and appeared to induce apoptotic form of cell death in many cells of SpIm culture.These results indicate that NPVs exhibit complex feature in the cells depending upon both the species of NPV and cultured cells being inoculated, and establish important basis for the analysis of molecular mechanisms of host range determination in NPVs.

将苜蓿银纹夜蛾（Ac）、家蚕（Bm）、美国白蛾（Hc）和甜菜夜蛾（Se）核多角体病毒（NPV）分别接种至SF21细胞、BmN-4细胞、SpIm细胞和Se301细胞，并观察细胞病变效应（CPE）表现、病毒 DNA 产生、进入培养基的病毒粒子产量、病毒对接种细胞中的结构蛋白产生、多角体蛋白（多面体的基质蛋白）产生和多面体形成进行了比较检查。结果表明，AcNPV 能够在所有四种检查的细胞系中繁殖并产生出芽病毒粒子，但多角体和多角体的产生仅发生在 SF21 细胞、SpIm 细胞和 Se301 细胞中，而不会在 BmN-4 细胞中产生。 BmN-4 细胞中的滴度和 SpIm 细胞中的滴度较低，其中只有少数细胞表现出 CPE。未观察到明显的 CPE在接种 BmNPV.HcNPV 的 SF21 和 Se301 细胞中，HcNPV 明显繁殖，并在 SpIm 和 Se301 细胞中产生多面体。在 SF21 细胞中，HcNPV 仅产生低滴度的出芽病毒粒子，并且既不产生多面体也不产生多面体。在接种 HcNPV 的 BmN-4 细胞中，积累了大量病毒 DNA，但从未检测到病毒结构蛋白和多角体蛋白的积累以及出芽病毒颗粒的产量。 SeNPV 仅在 Se301 细胞中明显增殖，并且似乎在 SpIm 培养物的许多细胞中诱导细胞凋亡形式的死亡。这些结果表明，NPV 在细胞中表现出复杂的特征，具体取决于 NPV 的种类和接种的培养细胞，并建立分析NPV宿主范围决定的分子机制的重要基础。