ALTERATIONS IN CNS FUNCTION AND DEVELOPMENT IN PEDIATRIC AIDS

建议的研究集中在细胞和分子基础上。 中枢神经系统(CNS)功能和发育的改变 发生在儿童艾滋病(HIV-1相关进行性脑病 童年)。感染人类免疫缺陷病毒(HIV)的儿童 展示了各种被认为是由艾滋病毒引起的神经系统疾病 中枢神经系统感染。这些异常的生物学基础是 未知，但可能涉及中枢神经功能、结构的变化 和发展。理解和处理此类问题的能力将是 对受感染的儿童和整个社会都具有重大价值。 我们的研究将集中在细胞因子和HIV外壳的可能作用上 这些异常中的蛋白质(如GP-120、GP-160、GP-41)。这个 待检测的细胞因子包括已知的IL-1β、TNF-α和IL-6 在感染艾滋病毒的人的大脑或脑脊液中含量升高 个人。这些细胞因子由巨噬细胞和小胶质细胞分泌， 中枢神经系统中携带病毒的细胞。细胞因子和细胞因子的作用 病毒蛋白将单独和联合进行研究，以确定 如果它们的影响是相加的或协同的。治疗条件 显著改变神经元的功能或发育将是 详细研究以确定改变的位置和潜在的机制 在分子水平上。细胞因子和病毒蛋白的影响将是 在培养的小脑颗粒神经元中进行研究，一种有利的 发展模式一直是我们过去研究的重点 资助期。许多重要的形态和生理里程碑 已知发生在体内这些神经元成熟过程中的基因表达 在培养中，因此可用于体外研究。神经元将会是 通过培养长期接触细胞因子和/或病毒蛋白 5~6成熟。这些因素对细胞形态和生理功能的影响 颗粒神经元的发育将用 免疫组织化学、电生理和显微钙成像 方法：研究方法。颗粒神经元将在同质神经元中生长。 培养或与星形胶质细胞共培养以确定细胞因子和/或 病毒蛋白可直接诱导颗粒神经元功能改变或 需要星形胶质细胞的干预。治疗剂将是 测试是否有能力改善TH细胞因子和/或 病毒蛋白。此外，转基因小鼠的颗粒神经元 过度表达的细胞因子和/或病毒蛋白将被检查是否发生改变 提供体外研究的体内相关性的功能 这些因素的外源应用。这些研究的结果应该是 提供有关神经元和分子机制的重要信息， 在儿童中观察到的认知和发育问题 感染了艾滋病毒。

The proposed studies focus on the cellular and molecular bases of alterations in central nervous system (CNS) function and development that occur in pediatric AIDS (HIV-1-associated progressive encephalopathy of childhood). Children infected with the human immunodeficiency virus (HIV) exhibit a variety of neurologic disorders thought to be caused by HIV infection in the CNS. The biological bases for these abnormalities are unknown, but presumably involve changes in CNS neuronal function, structure and development. The ability to understand and treat such problems will be of significant value to the infected children and to society as a whole. Our studies will focus on the possible role of cytokines and HIV coat proteins (e.g., gp- 120, gp- 160, gp-41) in these abnormalities. The cytokines to be tested include IL-1beta, TNF-alpha and IL-6 which are known to be present at elevated levels in the brains or CSF of HIV infected individuals. These cytokines are secreted by macrophages and microglia, the cells in the CNS that harbor the virus. The effects of cytokines and viral proteins will be investigated alone and in combination to determine if their effects are additive or synergistic. Treatment conditions that significantly alter neuronal function function or development will be studied in detail to identify the altered sites and underlying mechanisms at the molecular level. Effects of cytokines and viral proteins will be investigated in cerebellar granule neurons in culture, a favorable developmental model that has been the focus of our studies during the last funding period. Many important morphological and physiological milestones known to occur during the maturation of these neurons in vivo are expressed in culture and thus are accessible for study in vitro. The neurons will be exposed chronically to the cytokines and/or viral proteins via the culture medium. Effects of these factors on the morphological and physiological development of the granule neurons will be investigated using immunohistochemical, electrophysiological and microscopic calcium imaging methods. The granule neurons will be grown in homogeneous neuronal cultures or co-cultured with astrocytes to determine if cytokines and/or viral proteins can directly induce altered granule neuronal function or if the intervention of astrocytes is required. Therapeutic agents will be tested for the ability to ameliorate the effects of th cytokines and/or viral proteins. In addition, granule neurons from transgenic mice overexpressing cytokines and/or viral proteins will be examined for altered function to provide an in vivo correlate of the in vitro studies involving exogenous application of these factor. Results from these studies should provide important information on neuronal and molecular mechanisms that may contribute to the cognitive and developmental problems observed in children infected with HIV.