CNS INFLAMMATION IN NERVOUS AND MENTAL DISEASE

神经和精神疾病中的中枢神经系统炎症

• 批准号: 2392989
• 负责人: Joel S Pachter
• 金额: $ 24.01万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2392989
• 关键词: antibody; astrocytes; blood brain barrier; cell migration; central nervous system; chemoattractants; cytomegalovirus; electron microscopy; fluorescence microscopy; human tissue; inflammation; interferon gamma; interleukin 1; interleukin 2; leukocyte activation /transformation; lipopolysaccharides; monocyte; phorbols; tissue /cell culture; transforming growth factors; tumor necrosis factor alpha; vascular endothelium

Cells of the monocyte/macrophage lineage are now thought to be prominent effector cells that leave the circulation and play a pathogenetic role in several neurologic disorders that affect mental health status, including AIDS dementia complex, multiple sclerosis and Alzheimer's disease. However, little is known of the signals that promote the extravasation of monocytes through the restrictive blood-brain barrier (BBB), or the cells with which they interact once they enter the brain. To address these issues, a cell culture model of the human BBB will be used to analyze - for the first time - the distinct phases of monocyte migration through the BBB in response to a variety of stimuli. The long-term objective of this proposal is to test the hypothesis that monocyte migration across the BBB can be promoted by a variety of factors, e.g., specific cytokines, chemotaxins and other substances, which affect the activation state of either monocytes or brain microvessel endothelial cells (BMEC). Initial studies will assess the effects of monocyte stimulation/activation with lipopolysaccharide (LPS), phorbol myristate acetate (PMA) and interferon- gamma (IFN-gamma). These agents have been shown to promote monocyte adhesion to/migration across peripheral vasculature, but have yet to be analyzed with respect to monocyte interaction with the BBB. This is critical, as BBB endothelium may behave distinctively from that of peripheral vessels. Subsequent studies will analyze the role of endothelial stimulation/activation by LPS and cytokines tumor necrosis factor-alpha (TNF-alpha), IFN-gamma and interleukins-l and -2 (IL-1, IL-2) - substances again known to foster monocyte migration across the peripheral circulation, and which, in the case of the cytokines, have been shown to be up-regulated in the brain concurrent with monocytic infiltration. Combinations of monocyte/BMEC stimulation will also be performed to see if this further enhances monocyte migration. The effect of BMEC infection with cytomegalovirus (CMV) will be evaluated as well, as CMV infects BMEC in AIDS, and has been shown to augment monocyte adhesion to infected peripheral vessel endothelial cells. CMV may thus aid in recruiting monocytes to the brain. The effects of monocyte chemotactic peptides MCP-1 and TGF-beta will additionally be analyzed, as these may be released by astrocytes during CNS inflammatory disease, and could also foster monocyte trafficking to the brain. These studies will be performed in the presence and absence of astrocyte-conditioned media, as such media has been shown to influence BBB permeability and, consequently, glial- derived factors might influence monocyte migration. BBB integrity will be analyzed during the course of these studies by both electron and fluorescence microscopy, to determine whether monocyte migration requires or induces endothelial cell damage. Antibody "blocking" experiments will be also be performed to identify cell adhesion molecule:ligand pathways operant in monocyte migration across the BBB. Lastly, the effects of monocyte:astroglial interactions on monocyte migration across the BBB model will be evaluated as well. These experiments will illuminate factors that regulate monocyte entry into the brain, and thus identify potential targets for therapeutic intervention in a variety of crippling CNS diseases.

现在认为单核细胞/巨噬细胞谱系的细胞是突出的 效应细胞离开循环并在 影响心理健康状况的几种神经系统疾病，包括 艾滋病痴呆症复合物，多发性硬化症和阿尔茨海默氏病。 然而，对促进渗出的信号知之甚少 通过限制性血脑屏障（BBB）或细胞的单核细胞 他们进入大脑后与之相互作用。解决这些 问题，人类BBB的细胞培养模型将用于分析 - 第一次 - 单核细胞迁移的独特阶段 BBB响应各种刺激。这个长期目标 提案是测试单核细胞迁移在BBB的假设 可以通过多种因素（例如特定的细胞因子）来促进 趋化素和其他物质，这些物质影响激活状态 单核细胞或脑微血管内皮细胞（BMEC）。最初的 研究将评估单核细胞刺激/激活的影响 脂多糖（LPS），佛波尔肉豆蔻酸酯（PMA）和干扰素 - 伽玛（IFN-Gamma）。这些药物已被证明可以促进单核细胞 对跨外围脉管系统的粘附/迁移，但尚未 关于与BBB的单核细胞相互作用进行了分析。这是 批判性，因为BBB内皮可能与 外围容器。随后的研究将分析 LPS和细胞因子肿瘤坏死的内皮刺激/激活 因子-Alpha（TNF-Alpha），IFN-Gamma和interleukins-L和-2（IL-1，IL-2） - 再次已知可以促进单核细胞迁移的物质 周围循环，在细胞因子的情况下， 显示在与单核细胞并发的大脑中上调 浸润。单核细胞/BMEC刺激的组合也将是 表演以查看这是否进一步增强了单核细胞迁移。效果 还将评估用巨细胞病毒（CMV）感染的BMEC感染 CMV在艾滋病中感染了BMEC，并已显示可增强单核细胞粘附 感染的周围血管内皮细胞。 CMV因此可能有助于 将单核细胞募集到大脑。单核细胞趋化性的影响 肽MCP-1和TGF-beta将进一步分析，因为这些可能是 在中枢神经系统炎症性疾病期间由星形胶质细胞发布，也可能 促进单核细胞运输到大脑。这些研究将进行 在存在和不存在星形胶质细胞条件的介质的情况下，作为此类媒体 已显示会影响BBB的渗透性，从而影响Glial- 派生因素可能会影响单核细胞迁移。 BBB的完整性将是 在这些研究过程中，电子和 荧光显微镜，以确定单核细胞迁移是否需要 或诱导内皮细胞损伤。抗体“阻断”实验将 也可以进行识别细胞粘附分子：配体途径 单核细胞迁移跨BBB的操作。最后，影响 单核细胞：关于单核细胞迁移跨BBB的星形胶质相互作用 模型也将进行评估。这些实验将阐明因素 调节单核细胞进入大脑，从而确定潜力 在各种残废中枢神经系统中进行治疗干预的目标 疾病。