USE OF DI RNA VECTOR FOR STUDING VIRAL PATHOGENESIS

小鼠冠状病毒小鼠肝炎病毒(MHV)引起脑脊髓炎 以及啮齿动物的脱髓鞘。它已被用作 研究人类多发性硬化症。该提案的长期目标 研究的目的是阐明其分子机制。 MHV的神经发病机制。具体目标是重点关注的 新型MHV缺陷干扰(DI)RNA载体的研制 一种表达病毒基因的系统，这些基因编码尖峰和 伴随病毒感染的血凝素/酯酶蛋白，以及 确定这些蛋白在病毒神经发病机制中的作用。这个 系统也将被用来评估通过这些分子机制 病毒蛋白被结合到病毒颗粒中。此外, 细胞因子和细胞因子拮抗剂将通过DI RNA表达 载体及其在体内外对病毒复制的影响 被评估。表达蛋白的分布和 将评估中枢神经系统的组织病理学变化 通过对小鼠脑和脊髓切片进行免疫酶染色 使用针对蛋白质和组织学检查的特异性抗体。 这一载体系统将提供一种独特的病毒蛋白表达方式。 无需使用外源载体，并确保病毒的合成 通过与正宗病毒蛋白相同的机制获得蛋白质。 此外，S细胞因子等外源基因可以在当地表达 在中枢神经系统，因此，提供了一种直接评估 细胞因子或细胞因子拮抗剂在病毒感染中的作用。这个 这项研究的结果将为我们提供对 神经系统疾病的机制和病毒的一般致病机制。

Murine coronavirus mouse hepatitis virus (MHV) causes encephalomyelitis and demyelination in rodents. It has been used as a model system for studying human multiple sclerosis. The long-term goals of the propose investigation are to elucidate the molecular mechanism of neuropathogenesis of MHV. The specific objectives are focused on development of novel defective-interfering (DI) RNA of MHV as a vector system to express viral genes encoding the spike and the hemagglutinin/esterase proteins, concomitant with viral infection, and determining the role of these proteins in viral neuropathogenesis. The system will also be used to assess the molecular mechanism by which these viral proteins are incorporated into virus particles. In addition, cytokines and cytokine antagonists will be expressed via the DI RNA vector and their effects on viral replication in vitro and in vivo will be assessed. Distributions of the expressed proteins and histopathological changes of the central nervous system will be evaluated by immunoenzyme-staining of the sections of mouse brains and spinal cords using antibodies specific for the proteins and histological examinations. This vector system will provide a unique way of expressing viral proteins without the use of a foreign vector and assure the synthesis of viral proteins by the same mechanism as the authentic viral proteins. Furthermore, the foreign genes such s cytokines, can be locally expressed in the central nervous system, thus, providing a direct way of assessing the effects of cytokines or cytokine antagonists on viral infection. The results gained from this study will provide fundamental insights into the mechanism of neurological diseases and viral pathogenesis in general.