MECHANISMS OF RETROVIRUS-INDUCED NEURAL DYSFUNCTION

因为它们被确认为动物和更多的疾病病原体 最近对人类来说，现在很明显大多数逆转录病毒是 不同程度的嗜神经性。逆转录病毒诱导的机制 然而，目前对神经系统疾病的了解很少，而且 研究兴趣浓厚的课题。该项目旨在提供 为神经生物学和逆转录病毒学的研究培训提供足够的支持 为了使候选人史蒂文·W·道能够调查 逆转录病毒感染并改变神经细胞的功能。调查 提出将利用体外和体内系统和分子 克隆猫科逆转录病毒(猫科白血病病毒和猫科动物 免疫缺陷病毒(FIV))开发研究逆转录病毒的模型- 诱发的神经性疾病。 第一阶段工作的具体目标是：1)开发方法 猫中枢神经系统细胞的原代培养及鉴定 中枢神经系统(CNS)逆转录病毒感染的靶细胞；以及3) 使用体外和体内试验来鉴定和分析潜在的 嗜神经性逆转录病毒。将开发技术用于培养 三大类神经细胞的丰富种群： 星形胶质细胞、少突胶质细胞和神经元。免疫细胞化学技术 将用于鉴定培养的时间进程和敏感度 猫中枢神经系统细胞暴露后对猫逆转录病毒的体外感染 无论是无细胞病毒还是有病毒感染的单个核细胞。相关 体内研究将使用原位杂交和免疫细胞化学来 鉴定逆转录病毒感染猫中枢神经系统组织中的感染细胞。 潜在嗜神经性和/或神经毒力的猫科逆转录病毒株 将检查它们感染和/或诱导的不同能力 培养的猫中枢神经系统细胞的变化。 在第二阶段，这些调查将扩大到包括身份识别 中枢神经系统细胞表面受体介导逆转录病毒附着和 描述逆转录病毒引起的神经细胞功能的改变。 评估细胞功能变化的研究将包括 免疫细胞化学鉴定细胞表面和细胞表面蛋白表达的改变 感染神经胶质细胞的分化抗原，形态学研究 神经元的存活和生长特性，以及测量 脑下垂体激素分泌(脑垂体前叶细胞)。在活体研究中， 将在训练期间进行，将在体外和 猫科逆转录病毒体内趋神经性的研究 对培养的猫中枢神经系统细胞具有神经亲和性的细胞也能够诱导 猫脑池内接种后的神经系统疾病。

Since their identification as disease-causing agents of animals and more recently of humans, it is now apparent that most retroviruses are neurotropic to varying degrees. Mechanisms by which retroviruses induce neurologic disease are, however, currently both poorly understood and the subject of intense research interest. This project is intended to provide sufficient support for research training in neurobiology and retrovirology to enable the candidate, Steven W. Dow, to investigate mechanisms by which retroviruses infect and alter function of neural cells. The investigations proposed will utilize both in vitro and in vivo systems and molecularly cloned feline retroviruses (feline leukemia virus {FeLV} and feline immunodeficiency virus {FIV}) to develop models for study of retrovirus- induced neurologic disease. The specific aims of Phase 1 of the work are: 1) to develop methods for primary culture and characterization of feline CNS cells; 2) to identify central nervous system (CNS) target cells for retrovirus infection; and 3) to use in vitro and in vivo assays to identify and analyze potentially neurotropic retroviruses. Techniques will be developed for culture of enriched populations of the three major classes of neural cells: astrocytes, oligodendrocytes, and neurons. Immunocytochemical techniques will be used to identify the time course and susceptibility of cultured feline CNS cells to feline retrovirus infection in vitro after exposure to either cell-free virus or to virus-infected mononuclear cells. Correlative in vivo studies will use in situ hybridization and immunocytochemistry to identify infected cells in CNS tissues of retrovirus-inoculated cats. Potentially neurotropic and/or neurovirulent strains of feline retroviruses will be examined for their differential ability to infect and/or induce alterations in cultured feline CNS cells. In Phase 2, these investigations will be extended to include identification of CNS cell surface receptors that mediate retrovirus attachment and to characterize retrovirus-induced alterations in neural cell function. Studies to evaluate alterations in cell function will include immunocytochemical identification of altered expression of cell surface and differentiation antigens in infected glial cells, morphologic studies of survival and outgrowth characteristics of neurons, and measurement of pituitary hormone secretion (anterior pituitary cells). In vivo studies, to be ongoing during the training period, will correlate the in vitro and in vivo neurotropism of feline retroviruses by determining whether viruses that are neurotropic for cultured feline CNS cells are also able to induce neurologic disease in cats after intracisternal inoculation.