Toward a mouse model of HIV infection & drug addiction

描述：(申请人提供) 目前还没有小动物模型来研究药物滥用和 艾滋病毒感染背景下的神经发病机制。我们建议设立这样的 一种通过改造现有表达人CD4的转基因小鼠株而建立的模型 和我们的合作者D.Littman博士开发的CCR5或CXCR4。虽然 这些动物的hCD4/辅受体阳性T细胞被发现具有抵抗力 对HIV、转基因巨噬细胞和小胶质细胞，它们也表达这些 受体，没有进行测试。我们的结果和其他人的研究表明 原代啮齿动物巨噬细胞在体外可以高效复制HIV 通过人类的伪分型或表达绕过对病毒进入的阻止 感受器。因为巨噬细胞和小胶质细胞，而不是T细胞，是主要的 脑中HIV的靶标，表达人类受体的小鼠转基因 这些细胞可能复制HIV神经致病的重要参数。这个 此第一阶段CEBRA应用程序的总体目标是测试这种可能性。 第二阶段的研究将在药物方面进一步发展新模式。 虐待。具体目标有三个：1)病毒的确定和筛选 HIV在小鼠巨噬细胞和小胶质细胞中复制的细胞参数 体外比较不允许的小鼠淋巴细胞；2)测试转基因小鼠 用于脑和淋巴器官受体显示的hCD4和CCR5或CXCR4 并为小鼠巨噬细胞高效感染HIV创造条件 和体内的小胶质细胞；3)启动HIV感染转基因的评估 小鼠寻找HIV-1神经致病标志物。无论是常规的还是转基因的 小鼠细胞将在培养中被感染，用于病毒RNA和蛋白质的检测 生产、子代病毒表型和选定的细胞因子和趋化因子 表情。活体研究将测试人类小胶质细胞中CD4和辅助受体的显示， 巨噬细胞和淋巴细胞，并开始评估HIV感染 组织和脑内病毒DNA、RNA和蛋白质负荷的转基因小鼠 巨噬细胞/小胶质细胞与淋巴细胞，以及脑组织 免疫细胞化学。这些初步研究将作为 培育适合药物成瘾研究的小鼠品系的重大计划 在小鼠神经系统中产生艾滋病毒感染的背景下。

DESCRIPTION: (provided by the applicant) At present there is no small animal model for the study of drug abuse and neuropathogenesis in the context of HIV infection. We propose to establish such a model by adapting the existing transgenic mouse strains expressing human CD4 and CCR5 or CXCR4 developed by our collaborator Dr. D. Littman. Although hCD4/coreceptor positive T cells from these animals were found to be resistant to HIV, transgenic macrophages and microglial cells, which also express these receptors, were not tested. Our results and research by others indicate that primary rodent macrophages can efficiently replicate HIV in vitro once the block to virus entry is bypassed by pseudotyping or expression of human receptors. Since macrophages and microglia, and not T cells, are the primary targets for HIV in the brain, mouse transgenics expressing human receptors on these cells may reproduce important parameters of HIV neuropathogenesis. The overall goal of this Phase I CEBRA application is to test this possibility. Phase II studies will develop the new model further in the context of drug abuse. There are three Specific Aims: 1) To determine the viral and select cellular parameters of HIV replication in murine macrophages and microglia in vitro compared to nonpermissive murine lymphocytes; 2) To test mice transgenic for hCD4 and CCR5 or CXCR4 for receptor display in brain and lymphoid organs and to establish conditions for efficient HIV infection of murine macrophages and microglia in vivo; 3) To initiate evaluation of HIV infected transgenic mice for markers of HIV-1 neuropathogenesis. Both conventional and transgenic mouse cells will be infected in culture for assay of viral RNA and protein production, progeny virus phenotype, and selected cytokine and chemokine expression. In vivo studies shall test human CD4 and coreceptor display in microglia, macrophages, and lymphocytes and begin evaluation of HIV infection of transgenic mice by viral DNA, RNA, and protein burden in tissue and brain macrophages/microglia versus lymphocytes, and by brain tissue immunocytochemistry. These initial studies will serve as the foundation of a major program to generate mouse strains suitable for research in drug addiction in the context of productive HIV infection in the murine nervous system.