MECHANISMS OF RETROVIRUS-INDUCED NEURAL DYSFUNCTION

自从它们被鉴定为动物和其他动物的致病因子以来， 最近的人类，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 神经系统疾病，然而，目前都知之甚少， 研究兴趣浓厚的主题。 该项目旨在提供 为神经生物学和逆转录病毒学研究培训提供充分支持 让候选人史蒂文·W·道，调查机制， 逆转录病毒感染并改变神经细胞的功能。 调查 所提出的将利用体外和体内系统， 克隆的猫逆转录病毒（猫白血病病毒{FeLV}和猫 免疫缺陷病毒（FIV）开发研究逆转录病毒的模型- 诱发神经系统疾病。 第一阶段工作的具体目标是：（1）制定方法， 猫CNS细胞的原代培养和表征; 2）鉴定 逆转录病毒感染的中枢神经系统（CNS）靶细胞;和3） 使用体外和体内试验来识别和分析潜在的 嗜神经性逆转录病毒 将开发培养 三种主要神经细胞的富集群体： 星形胶质细胞、少突胶质细胞和神经元。 免疫细胞化学技术 将用于确定培养的时间进程和敏感性 猫CNS细胞暴露于体外猫逆转录病毒感染后， 无细胞病毒或病毒感染的单核细胞。 相关 体内研究将使用原位杂交和免疫细胞化学， 鉴定接种逆转录病毒的猫的CNS组织中的感染细胞。 猫逆转录病毒的潜在嗜神经性和/或神经毒性毒株 将检查它们感染和/或诱导 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识别 中枢神经系统细胞表面受体介导逆转录病毒附着， 表征逆转录病毒诱导的神经细胞功能改变。 评估细胞功能改变的研究将包括 细胞表面表达改变的免疫细胞化学鉴定， 分化抗原在感染的神经胶质细胞，形态学研究， 神经元的存活和生长特征，以及 垂体激素分泌（垂体前叶细胞）。 体内研究， 在培训期间持续进行，将使体外和 猫逆转录病毒的体内嗜神经性 对培养的猫CNS细胞具有亲神经性也能够诱导 脑池内接种后猫的神经系统疾病。

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.