THE ROLE OF CYTOKINES IN THE PATHOGENESIS OF AIDS DEMENTIA COMPLEX

细胞因子在艾滋病痴呆症发病机制中的作用

• 批准号: 6111527
• 负责人: IAIN Leslie CAMPBELL
• 金额: $ 0.44万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-15 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6111527
• 关键词: AIDS dementia complex; AIDS therapy; HIV envelope protein; HIV infections; RNase protection assay; antiinflammatory agents; antioxidants; astrocytes; cellular pathology; central nervous system; confocal scanning microscopy; cytokine; disease /disorder model; gene expression; genetically modified animals; histopathology; hypothalamic pituitary adrenal axis; in situ hybridization; interferon alpha; laboratory mouse; molecular pathology; neuroimmunomodulation; neuroprotectants; nonhuman therapy evaluation

HIV-1 infection is commonly accompanied by injury to the central nervous system. Affected individuals frequently exhibit serious, progressive behavioral and neurological complications. Although the precise molecular and cellular pathogenetic processes associated with HIV encephalopathy remain unknown, accumulating evidence suggest toxicity tot he CNS amy be mediated indirectly, in part, by host derived factors such as cytokines, produced in response to the viral infection. This proposal focuses on the hypothesis that a key antiviral cytokine IFN-alpha, produced systemically and by infiltrating immune cells or resident brain cells, contributes to CNS injury during HIV-infection. To test this hypothesis, a well defined transgenic approach was employed in which the expression of IFN-alpha was targeted to astrocytes using a glial fibrillary acidic protein-IFNalpha1 fusion gene construct. This approach has provided a unique and powerful model to study the neuropathogenic consequences of the constitutive production of IFN-alpha from astrocytes in the intact CNS. Preliminary pathological characterization of so-called GIFN transgenic mice has unveiled wide-ranging structural and molecular alterations of the CNS thereby directly supporting the view that IFN-alpha, and likely therefore, other members of the IFN family, may have a causal role in the genesis of HIV encephalopathy. Here, we propose to further expand the scope and detail of the neuropathological assessment in an existing as well as in new stable lines of GIFN mice to be developed. This assessment will employ an established battery of tests to examine CNS alterations the molecular and cellular levels., including RNase protection assay, 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 GIFN mice will be determined at the behavioral, electrophysiological and neuroendocrine levels and where possible be linked to specific molecular and cellular alterations. The neurological impact of different pathogenetic factors will be assessed in the GIFN mice by: i) cross-breeding of the GFAP-IFN mice with mice from other transgenic backgrounds (e.g. GFAP-IL6 or GFAP-gp120), ii) intra- neural infection with neurotropic viruses and, iii) back-cross breeding with SCID mice to develop immunodeprived GFAP-cytokine transgenic animals. These studies will recapitulate the multi-factorial nature of the pathogenetic process the thought to underlie HIV encephalopathy. Finally, the well characterized GIFN mice will be used to identify and assess in vivo the efficacy of drugs targeted at harmful IFN-CNS interactions. This study provides a unique and powerful approach to elucidate the molecular and cellular basis for the CNS pathobiology of IFN-alpha in vivo and will facilitate the development and testing of therapeutic strategies to alleviate the toxic CNS actions of this cytokine.

HIV-1感染通常伴有中枢神经损伤 系统。 受影响的个体经常表现出严重的、进行性的 行为和神经系统并发症。 虽然精确的分子 和与 HIV 脑病相关的细胞发病过程 仍然未知，越来越多的证据表明对中枢神经系统有毒性 部分由宿主衍生因子（例如细胞因子）间接介导， 是针对病毒感染而产生的。 该提案的重点是 假设系统产生一种关键的抗病毒细胞因子 IFN-α 通过渗透免疫细胞或常驻脑细胞，有助于 HIV 感染期间的中枢神经系统损伤。 为了验证这个假设，需要定义一个明确的 采用转基因方法，其中 IFN-α 的表达 使用神经胶质纤维酸性蛋白 - IFNα1 靶向星形胶质细胞 融合基因构建体。 这种方法提供了一种独特而强大的 研究构成性神经病理后果的模型 完整 CNS 中的星形胶质细胞产生 IFN-α。 初步的 所谓 GIFN 转基因小鼠的病理特征 揭示了中枢神经系统的广泛结构和分子改变 从而直接支持了 IFN-α 的观点，并且很可能因此， IFN 家族的其他成员，可能在 IFN 的发生中具有因果作用 艾滋病毒脑病。 在此，我们建议进一步扩大范围 现有和新的神经病理学评估的细节 待开发的 GIFN 小鼠稳定系。 本次评估将采用 建立了一系列测试来检查中枢神经系统的分子和 细胞水平，包括 RNase 保护测定、原位杂交、 Northern印迹杂交、蛋白质免疫印迹测定、常规光 免疫标记脑切片和电子显微镜的激光共聚焦显微镜 显微镜。 将确定 GIFN 小鼠中枢神经系统功能的改变 在行为、电生理和神经内分泌水平上 可能与特定的分子和细胞改变有关。 这 将评估不同致病因素对神经系统的影响 GIFN 小鼠通过以下方式进行： i) GFAP-IFN 小鼠与来自以下小鼠的杂交 其他转基因背景（例如 GFAP-IL6 或 GFAP-gp120），ii) 内 嗜神经病毒的神经感染，以及，iii) 回交育种 与 SCID 小鼠一起开发免疫剥夺的 GFAP 细胞因子转基因动物。 这些研究将概括出影响因素的多因素性质。 发病过程被认为是艾滋病毒脑病的基础。 最后， 充分表征的 GIFN 小鼠将用于识别和评估 体内针对有害的 IFN-CNS 相互作用的药物的功效。 这 研究提供了一种独特而有力的方法来阐明分子 体内 IFN-α 的中枢神经系统病理学和细胞基础 促进治疗策略的开发和测试 减轻这种细胞因子的毒性中枢神经系统作用。