HIV INFECTION OF HUMAN FETAL NEURAL XENOGRAFTS

人类免疫缺陷病毒1型(HIV-1)感染中枢神经系统 成人和儿童的系统。人们对艾滋病病毒感染知之甚少。 胎儿神经系统，或关于母亲HIV-1的间接影响 对发育中的胎儿大脑的感染。目前还没有体外培养的 提供能满足负责人要求的系统或相关动物模型 解决致病问题或测试抗病毒药物的要求 可能治疗HIV-1脑部感染。标记的寄主物种专一性 慢病毒和病毒-宿主细胞相互作用的重要性表明 需要一个模型，在这个模型中完整的，分化的，人类胎儿 神经组织可能感染HIV-1和/或暴露在潜在的有毒物质中 病毒或宿主来源的因子。我们开发了一种模型，在这种模型中，人类 妊娠中期的胎儿大脑或视网膜组织作为异种移植生长在 成年大鼠的眼前房。这些异种移植物 血管化，建立血脑屏障，分化为 星形胶质细胞、小胶质细胞和神经元成分。 我们建议建立和优化感染这些人的条件 携带HIV-1病毒的胎儿神经组织。建议使用电池进行实验 游离病毒、病毒感染的淋巴细胞和巨噬细胞以及胎儿神经 移植前体外感染的外植体(特定目标1)。 确定细胞定位并分析其发生的过程。 在这些异种移植物中感染HIV-1，我们将使用聚合酶链式反应 (PCR)，结合免疫组织化学和原位杂交 技巧。这将确定哪些细胞类型(神经外胚层或其他) 被感染，以及每种细胞类型中感染的性质(生产性， 受限的或潜在的)。补充性研究将使用 分离神经元、星形胶质细胞和小胶质细胞成分的体外方法， 然后重新聚集这些细胞的特定组合，在 异种移植，作为HIV-1感染的靶点。(具体目标2) 我们将评估使用该模型评估 针对HIV-1感染的抗病毒药物的疗效和毒性 神经系统(具体目标3)。 最后，我们将使用异种移植模型来确定对 寄主细胞或寄主的假定毒性因子的神经系统发育 病毒来源(特定目标4)。

Human immunodeficiency virus type 1 (HIV-1) infects the central nervous system of adults and children. Little is known about HIV-1 infection of the fetal nervous system, or about the indirect effects of maternal HIV-1 infection on the developing fetal brain. There is currently no in vitro system or related animal model available which can satisfy the chief requirements to address pathogenetic Issues or to test anti-viral agents which may treat HIV-1 brain infection. The marked host species specificity of lentiviruses and the importance of viral-host cell interactions suggest that a model is required in which intact, differentiating, human fetal neural tissue can be infected by HIV-1 and/or exposed to potentially toxic viral or host derived factors. We have developed a model in which human second trimester fetal brain or retinal tissues are grown as xenografts in the anterior chamber of the eye in adult rats. These xenografts vascularize, establish a blood-brain-barrier, and differentiate into astroglial, microglial, and neuronal elements. We propose to establish and optimize conditions for infecting these human fetal neural tissues with HIV-1. Experiments are proposed employing cell free virus, virus infected lymphocytes and macrophages, and fetal neural explants infected in vitro prior to transplantation (Specific Aim 1). To determine the cellular localization and to analyze the course of the HIV-1 infection in these xenografts, we will use polymerase chain reaction (PCR), coupled with immunohistochemical and in situ hybridization techniques. This will identify which cell types (neuroectodermal or other) are infected, and the nature of Infection in each cell type (productive, restricted, or latent). Complementary studies will be performed using in vitro methods to segregate neuronal, astroglial, and microglial components, followed by re-aggregation of specific combinations of these cells, in the xenograft, to serve as a target for HIV-1 infection. (Specific Aim 2) We will evaluate the feasibility of of using this model to assess the efficacy and toxicity of antiviral agents which target HIV-1 infection of the nervous system (Specific Aim 3). Lastly we will use the xenograft model to determine the indirect effects on the developing nervous system of putative toxic factors of host cell or viral origin (Specific Aim 4).