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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（Autographa californica，Ac）、家蚕（Bmx mori，Bm）、美国白蛾（Hyphantria cunea，Hc）和甜菜夜蛾（Spodoptera exigua，Se）核型多角体病毒（Nucleopolyhedrovirus，NPVs）分别接种SF 21细胞、BmN-4细胞、SpIm细胞和Se 301细胞，观察细胞病变（CPE）表现、病毒DNA产生、病毒粒子加入培养液中的量、病毒结构蛋白产生、多角体蛋白（Polyhedrin）（多角体的基质蛋白）的产生和接种细胞中的多角体形成进行了比较检查。结果表明，AcNPV在4种细胞系中都能增殖并产生出芽病毒粒子，但多角体蛋白和多角体的产生仅见于SF 21细胞、SpIm细胞和Se 301细胞，BmNPV在BmN-4细胞中增殖到高滴度，而在BmN-4细胞中则没有。接种BmNPV的SF_（21）和Se_（301）细胞无明显的CPE，接种HcNPV的细胞数增加在SpIm和Se 301细胞中产生了多角体。在SF 21细胞中，HcNPV仅产生低滴度的出芽病毒体，并且既不产生多角体蛋白也不产生多角体。在接种HcNPV的BmN-4细胞中，病毒DNA大量积累，但病毒结构蛋白和多角体蛋白的积累和出芽病毒粒子产量从未检测到。SeNPV仅在Se 301细胞中增殖明显，在SpIm培养的许多细胞中出现诱导细胞凋亡形式的死亡，这些结果表明NPVs在细胞中表现出复杂的特征，这取决于接种的细胞和NPV的种类，为分析NPVs寄主范围决定的分子机制奠定了重要基础。