HIV INFECTION OF NEURAL TISSUE--AN ORGANOTYPIC MODEL

人胎儿中枢神经系统器官型培养 组织将被用来测试假设，人类 免疫缺陷病毒（HIV）可以在 怀孕 这种感染可能会导致 在一些先天性艾滋病病毒感染的儿童中观察到的功能障碍， 血清阳性、获得性免疫缺陷综合征（AIDS），或 艾滋病相关综合征（ARC）。 实验策略将 重点关注从择期妊娠中获得的胎儿CNS组织 终止艾滋病毒血清反应阴性的女性妊娠， 增加，以及来自艾滋病毒血清阳性或患有ARC或 艾滋病 HIV在器官型CNS中诱导病理的能力 将通过显微镜和免疫细胞化学评估培养物 结合核酸和蛋白质生物化学。 具体来说，艾滋病毒感染不同神经细胞的能力 类型及其在单元格中的位置将由 病毒颗粒和免疫电镜 HIV特异性蛋白的显微镜染色。 由于艾滋病毒是一种 逆转录病毒及其基因组或其部分可以被掺入 宿主细胞DNA，Southern印迹分析和原位核酸 将使用酸杂交来检验这一点。 一 HIV引起的神经病理学的一个组成部分可能涉及改变 在正常细胞类型的特异性蛋白质，因此，蛋白质印迹 分析将用于检查蛋白质的可能变化， 分化的合成和翻译后修饰 标记。 为了确定代谢的改变，另外 研究将集中在受感染组织中的酶动力学。 使用来自HIV感染者的流产培养物的类似实验 女性可以定义经胎盘胎儿感染的动力学 以及由此产生的神经病理学。 这些实验将增强 了解艾滋病毒感染的机制， 干扰正常CNS组织发育。 除了 确定：（1）HIV是否可以感染神经细胞;（2）哪些细胞 感染;（3）细胞结构和 功能可能由感染引起。 这些环境可能 定义HIV受体而不是T4分子。 这样的见解 可以允许设计改进的预防性或治疗性药物 防治艾滋病毒感染的议定书。

Organotypic cultures of human fetal central nervous system (CNS) tissue will be used to test the hypothesis that human immunodeficiency virus (HIV) can infect neural tissue during gestation. Such an infection might account for the neurologic dysfunction seen in some children with congenital HIV- seropositivity, acquired immunodeficiency syndrome (AIDS), or AIDS-related complex (ARC). The experimental strategies will focus on fetal CNS tissue obtained from elective pregnancy terminations of HIV-seronegative females, to which HIV will be added, and from females who are HIV-seropositive or have ARC or AIDS. The ability of HIV to induce pathology in organotypic CNS cultures will be assessed by microscopy and immunocytochemistry in conjunction with nucleic acid and protein biochemistry. Specifically, the ability of HIV to infect different neural cell types and its location within cells will be determined by electron microscopy for viral particles and immunoelectron microscopic staining for HIV-specific proteins. Since HIV is a retrovirus and its genome, or parts thereof, may be incorporated into host cell DNA, Southern Blot analysis and in situ nucleic acid hybridization will be used to examine this point. A component of HIV-induced neuropathology may involve alterations in normal cell-type specific proteins, therefore, Western Blot analyses will be used to examine possible changes in protein synthesis and post-translational modifications of differentiation markers. To determine alterations in metabolism, additional studies will focus on enzyme kinetics in infected tissue. Similar experiments using abortus cultures from HIV-infected females may define the kinetics of transplacental fetal infection and the resultant neuropathology. These experiments will enhance the understanding of the mechanisms by which HIV infection may interfere with normal CNS tissue development. In addition to determining: (1) if HIV can infect neural cells; (2) which cells are infected; and, (3) what perturbations of cell structure and function may result from infection. Those environments may define HIV receptors other than the T4 molecule. Such insights may permit the design of improved preventative or therapeutic protocols for combatting HIV infection.