USE OF DI RNA VECTOR FOR STUDING VIRAL PATHOGENESIS

鼠冠状病毒小鼠肝炎病毒（MHV）引起脑脊髓炎 和脱髓鞘。 它已被用作模型系统， 研究人类多发性硬化症 建议的长期目标 研究的目的是阐明 MHV的神经发病机制。 具体目标集中在 新型MHV缺陷干扰（DI）RNA载体的开发 系统，以表达编码刺突的病毒基因， 血凝素/酯酶蛋白，伴随病毒感染，和 确定这些蛋白质在病毒神经发病机制中的作用。 的 系统也将被用来评估分子机制，这些 病毒蛋白被掺入病毒颗粒中。 此外，本发明还提供了一种方法， 细胞因子和细胞因子拮抗剂将通过DI RNA表达 载体及其在体外和体内对病毒复制的作用将 进行评估。 表达蛋白的分布和 将评价中枢神经系统的组织病理学变化 通过小鼠脑和脊髓切片的免疫酶染色 使用蛋白质特异性抗体和组织学检查。 这种载体系统将提供一种表达病毒蛋白的独特方式 不使用外源载体，并确保病毒的合成， 通过与真正的病毒蛋白相同的机制。 此外，外源基因如细胞因子可在细胞内局部表达， 在中枢神经系统中，因此，提供了一种直接的方法来评估 细胞因子或细胞因子拮抗剂对病毒感染的影响。 的 从这项研究中获得的结果将提供基本的见解， 神经系统疾病的发病机制和一般的病毒发病机制。

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.