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CYTOKINES AND AIDS DEMENTIA COMPLEX

细胞因子和艾滋病痴呆症

基本信息

批准号：
6151443
负责人：
IAIN Leslie CAMPBELL
金额：
$ 45.41万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-02-01 至 2002-01-31
项目状态：
已结题

项目摘要

This proposal focuses on the hypothesis that cytokines produced systemically and by infiltrating immune cells or resident brain cells, contribute to CNS injury during HIV-infection. To test this hypothesis, a well defined transgenic approach was employed in which the expression of the cytokines IL-6 and IL-3 was targeted to astrocytes using glial fibrillary acidic protein (GFAP)-fusion gene constructs. This has provided us with unique and powerful models to study the neuropathogenic consequences of the constitutive production of cytokines from astrocytes in the intact CNS. Initial characterization of GFAP-IL6 and GFAP-IL3 transgenic mice has unveiled wide-ranging molecular, cellular and functional alterations of the CNS-many of which share similarities to those seen in HIV encephalopathy. Significantly, these studies directly implicate cytokines in having a causal role in the genesis of HIV encephalopathy and other neurodegenerative diseases. Here we propose to develop transgenic mice with expression of the cytokine TNF-alpha targeted to the CNS. Detailed neuropathological assessment in this new model as well as in existing GFAP-cytokine mice will employ an established battery of tests to examine CNS alterations at the molecular and cellular levels, including RNase protection assays, in situ hybridization, northern blot hybridization, protein immunoblot assay, conventional light and laser confocal microscopy of immunolabeled brain sections and electron microscopy. Functional CNS alterations in the GFAP- cytokine mice will be determined at the behavioral and electrophysiological and levels and where possible be linked to specific molecular and cellular alterations. The identification of primary pathogenetic and functional milestones associated with the cerebral expression of the various cytokines will be determined by: i) detailed developmental studies and comparative analysis of the different GFAP- cytokine models, and ii) analyzing the CNS alterations resulting from the grafting of cytokine producing transgenic astrocytes in the normal mouse brain. The neurological impact of additional pathogenetic factors will be assessed: i) in cross-breeding experiments to develop biogenic mice expressing combinations of cytokines (i.e. IL-6+IL-3), and ii) by back- cross breeding GFAP-cytokine mice with SCID mice to develop immunodeficient GFAP-cytokine transgenic animals. These studies will develop models that recapitulate the multi-factorial pathogenetic and immunodeficient environments thought to underlie HIV encephalopathy. Finally, the well characterized GFAP-transgenic mice will be used to identify and assess in vivo the efficacy of drugs targeted at harmful individual cytokine-CNS interactions. This study provides a unique and powerful approach to elucidate the molecular and cellular basis for the CNS pathobiology of cytokines in vivo and can be expected to advance our understanding of HIV-associated neurological disease, help identify critical targets for therapeutic interventions and facilitate the preclinical evaluation of therapeutic strategies.

这项建议的重点是假设，细胞因子产生全身性地和通过浸润免疫细胞或常驻脑细胞，导致HIV感染期间CNS损伤。为了验证这个假设，采用了一种明确的转基因方法，细胞因子IL-6和IL-3使用胶质细胞靶向星形胶质细胞，胶质酸性蛋白（GFAP）-融合基因构建体。这为我们提供了独特而强大的模型来研究神经致病性从星形胶质细胞组成性产生细胞因子的后果完整的中枢神经系统GFAP-IL6和GFAP-IL3的初步表征转基因小鼠已经揭示了广泛的分子，细胞和中枢神经系统的功能改变-其中许多与在HIV脑病中看到的。这些研究直接暗示细胞因子在HIV的发生中具有因果作用脑病和其他神经变性疾病。在此，我们建议开发表达细胞因子TNF-α的转基因小鼠针对CNS。详细的神经病理学评估，在这个新的模型以及现有的GFAP-细胞因子小鼠将采用建立了一系列测试，以检查分子水平上的CNS改变，和细胞水平，包括RNA酶保护试验，原位杂交、北方印迹杂交、蛋白质免疫印迹分析免疫标记脑的常规光和激光共聚焦显微镜切片和电子显微镜检查。GFAP中的功能性CNS改变- 细胞因子小鼠将在行为和电生理学和水平，并在可能的情况下与特定的分子和细胞的改变。识别主要与脑梗死相关的发病机制和功能里程碑各种细胞因子的表达将通过以下方式确定：i）详细描述不同GFAP的发展研究和比较分析，细胞因子模型，和ii）分析由所述细胞因子模型引起的CNS改变。细胞因子转基因星形胶质细胞在正常小鼠体内的移植个脑袋其他致病因素的神经影响将 i）在杂交育种实验中开发生物基因小鼠表达细胞因子的组合（即IL-6 + IL-3），和ii）通过回- 将GFAP-细胞因子小鼠与SCID小鼠杂交以开发免疫缺陷GFAP-细胞因子转基因动物。这些研究将开发概括多因素致病因素的模型，免疫缺陷环境被认为是HIV脑病的基础。最后，良好表征的GFAP转基因小鼠将用于在体内确定和评估针对有害生物的药物的疗效单独的精氨酸-CNS相互作用。这项研究提供了一个独特的，强大的方法来阐明分子和细胞的基础， CNS病理生物学的细胞因子在体内，可以预期，以推进我们的研究。了解艾滋病毒相关的神经系统疾病，治疗干预的关键目标，并促进治疗策略的临床前评价。