CNS INFLAMMATION IN NERVOUS AND MENTAL DISEASE

单核/巨噬细胞系的细胞现在被认为是突出的 离开血液循环并发挥致病作用的效应细胞 影响精神健康状况的几种神经障碍，包括 艾滋病痴呆症、多发性硬化症和阿尔茨海默病。 然而，人们对促进血管外渗的信号知之甚少。 通过限制性血脑屏障(BBB)的单核细胞 一旦它们进入大脑，就会与之相互作用。要解决这些问题 问题，将使用人类血脑屏障的细胞培养模型来分析- 第一次-单核细胞迁移的不同阶段 血脑屏障对各种刺激的反应。这样做的长期目标是 建议检验单核细胞跨血脑屏障迁移的假设 可由多种因素促进，例如，特定的细胞因子， 趋化因子和其他物质，它们影响细胞的激活状态 单核细胞或脑微血管内皮细胞(BMEC)。首字母 研究将评估单核细胞刺激/激活的效果 脂多糖、佛波酯和干扰素- 伽玛(干扰素-伽马)。这些药物已被证明能促进单核细胞 与周围血管系统的黏附/迁移，但尚未发生 分析单核细胞与血脑屏障的相互作用。这是 关键，因为血脑屏障内皮细胞的行为可能与 周围血管。后续的研究将分析 内毒素刺激/激活内皮细胞与细胞因子肿瘤坏死 肿瘤坏死因子-α、干扰素-γ和白介素2-L、白介素2 -再次已知可促进单核细胞在世界各地迁移的物质 外周循环，在细胞因子的情况下，这是 显示与单核细胞同时在大脑中上调 渗透。单核细胞/骨髓间充质干细胞刺激组合也将是 以查看这是否进一步增强了单核细胞的迁移。其效果 还将评估BMEC感染巨细胞病毒(CMV)的情况，以及 巨细胞病毒可感染艾滋病患者的BMEC，并已被证明可增强单核细胞的黏附 感染的外周血管内皮细胞。因此，CMV可能会帮助 向大脑招募单核细胞。单核细胞趋化作用研究进展 多肽MCP-1和转化生长因子-β将被额外分析，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释放，也可以 助长单核细胞向脑内贩运。这些研究将被执行 在存在和不存在星形胶质细胞条件培养液的情况下，如这种培养液 已被证明影响血脑屏障的通透性，因此，胶质细胞- 衍生因素可能影响单核细胞的迁移。BBB的完整性将是 在这些研究过程中分析了电子和 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“封闭”实验将 BE也可用于识别细胞黏附分子：配体途径 在单核细胞跨血脑屏障迁移中起作用。最后，它的影响 单核细胞：星形胶质细胞相互作用对单核细胞跨血脑屏障迁移的影响 模型也将被评估。这些实验将阐明各种因素 调节单核细胞进入大脑，从而识别潜在的 多种致残中枢神经系统的治疗干预靶点 疾病。

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.