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: 6928522
• 负责人: Joel S Pachter
• 金额: $ 21.75万
• 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 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6928522
• 关键词: antibody; astrocytes; blood brain barrier; cell migration; central nervous system; cytomegalovirus; electron microscopy; fluorescence microscopy; human tissue; inflammation; interferon gamma; interleukin 1; interleukin 2; leukocyte activation /transformation; lipopolysaccharides; monocyte; monocyte chemoattractant protein 1; phorbols; tissue /cell culture; transforming growth factors; tumor necrosis factor alpha; vascular endothelium

EXCEED THE SPACE PROVIDED. The chemokines monocyte chemoattractant protein (MCP-1) and macrophage inflammatory protein-1 alpha (MlP-la) are regularly observed in the brain during episodes of inflammation, and thus, are thought to be critical effectors in directing leukocyte infiltration into the central nervous system (CNS). Nevertheless, no direct evidence exists that either one of these two chemokines can actually stimulate leukocyte migration through the highly-impermeable blood-brain barrier (BBB). Moreover, it is completely unclear how these chemokines. which derive mainly from perivascular glia and extravasated leukocytes, exert their actions across the BBB. In light of the profound impact these substances could have on the course and treatment of neuroinflammatory disease, it is crucial that these issues be resolved. Toward this end, experiments are proposed to test the following hypothesis: Chemokines MCP-1 and MIP1- a stimulate mononuclear migration across the BBB, and do so by specific interactions with brain microvascular endothelial cells. Specifically, experiments will be aimed at clarifying the following issues: 1) Does MCP-1 or MIPl-cc, alone, have the capacity to stimulate transendothelial migration across the BBB. or does each need to work in concert with other proinflammatory cytokines? 2) By what mechanisms(s) might these chemokinesrelay their signals across the BBB? Initial experiments will use an in vitro model of the human BBB. developed in this laboratory, to directly measure the effects of MCP-1 and MlP-la on monocyte transendothelial migration. The next series of studies will utilize both this model and isolated human brain microvessels to investigate whether these chemokines are transported paracellularly or transcellularly across the specialized brain microvascular endothelium. Subsequent investigations will determine whether recognized receptors for these chemokines exist on the abluminal microvascular surface, and,if so, to examine their role in chemokine transport. The final experiments will illuminate microvascular responses to chemokine stimulation, e.g..altered adhesion molecule expression and permeability, which could support leukocyte infiltration. This study will be the first to characterize the interactions of MCP-1 and MlP-la at the BBB. Furthermore, in appreciation of the critical role of MCP-1 and MlP-la in CNS inflammation, these experimentswill greatly aid in comprehending pathogenetic mechanisms involved in this condition, and in developing novel therapeutics to antagonize chemokine actions in neuroinflammatorydisease. PERFORMANCE SITE ========================================Section End===========================================

超出所提供的空间。在炎症发作期间，在大脑中经常观察到趋化因子单核细胞趋化蛋白 (MCP-1) 和巨噬细胞炎症蛋白 1 α (MIP-la)，因此，它们被认为是引导白细胞浸润到中枢神经系统 (CNS) 的关键效应因子。然而，没有直接证据表明这两种趋化因子中的任何一种实际上可以刺激白细胞通过高度不可渗透的血脑屏障（BBB）迁移。而且，目前还完全不清楚这些趋化因子是如何发挥作用的。主要源自血管周围的神经胶质细胞和外渗的白细胞，在血脑屏障上发挥作用。鉴于这些物质可能对神经炎症性疾病的病程和治疗产生深远影响，解决这些问题至关重要。为此，提出实验来检验以下假设：趋化因子 MCP-1 和 MIP1-a 刺激单核细胞跨 BBB 迁移，并通过与脑微血管内皮细胞的特定相互作用来实现这一点。具体而言，实验旨在澄清以下问题：1)MCP-1或MIP1-cc单独是否具有刺激穿过BBB的跨内皮迁移的能力。还是每种细胞因子都需要与其他促炎细胞因子协同作用？ 2) 这些趋化因子通过什么机制在血脑屏障上传递信号？最初的实验将使用人类血脑屏障的体外模型。该实验室开发的直接测量MCP-1和MLP-la对单核细胞跨内皮迁移的影响。下一系列研究将利用该模型和分离的人脑微血管来研究这些趋化因子是否以旁细胞或跨细胞方式穿过专门的脑微血管内皮。随后的研究将确定这些趋化因子的公认受体是否存在于腔外微血管表面，如果存在，则检查它们在趋化因子转运中的作用。最终的实验将阐明微血管对趋化因子刺激的反应，例如改变粘附分子的表达和渗透性，这可以支持白细胞浸润。这项研究将首次描述 MCP-1 和 MLP-la 在 BBB 上的相互作用。此外，由于认识到MCP-1和MIP-1a在中枢神经系统炎症中的关键作用，这些实验将极大地有助于理解与这种情况有关的发病机制，并开发新的疗法来拮抗神经炎症疾病中的趋化因子作用。表演网站==========================================章节结束===============================================

项目成果

Analysis of mouse brain microvascular endothelium using laser capture microdissection coupled with proteomics.

使用激光捕获显微切割结合蛋白质组学分析小鼠脑微血管内皮。

• DOI: 10.1007/978-1-60761-938-3_14
• 发表时间: 2011
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Murugesan,Nivetha;Macdonald,JenniferA;Lu,Qiaozhan;Wu,Shiaw-Lin;Hancock,WilliamS;Pachter,JoelS
• 通讯作者: Pachter,JoelS

Monocyte chemoattractant protein-1 alters expression of tight junction-associated proteins in brain microvascular endothelial cells.

• DOI: 10.1016/j.mvr.2003.07.001
• 发表时间: 2004
• 期刊: Microvascular research
• 影响因子: 3.1
• 作者: Li Song;J. Pachter

Probing the CNS microvascular endothelium by immune-guided laser-capture microdissection coupled to quantitative RT-PCR.

通过免疫引导激光捕获显微切割结合定量 RT-PCR 探测中枢神经系统微血管内皮。

• DOI: 10.1007/978-1-61779-163-5_32
• 发表时间: 2011
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Murugesan,Nivetha;Macdonald,Jennifer;Ge,Shujun;Pachter,JoelS
• 通讯作者: Pachter,JoelS