DYMANICS OF ENDOTHELIAL CELL SIGNALING AND SIVE NEUROINFLAMMATION

内皮细胞信号传导和严重神经炎症的动力学

• 批准号: 8173036
• 负责人: ANDREW G MACLEAN
• 金额: $ 6.18万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173036
• 关键词: Actins; Astrocytes; Blood - brain barrier anatomy; Brain; CCL2 gene; Computer Retrieval of Information on Scientific Projects Database; Confocal Microscopy; Cytoskeletal Modeling; Down-Regulation; Endothelial Cells; Endothelium; Functional disorder; Funding; Grant; IL8 gene; Infection; Inflammation; Inflammatory; Institution; Ligands; MYLK gene; Macaca; Mediating; Messenger RNA; Neuraxis; PTK2 gene; Pathway interactions; Permeability; Phosphorylation; Production; RNA; Reaction; Regulation; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; SIV; Signal Transduction; Source; Stimulus; TNF gene; TNFSF10 gene; Tight Junctions; United States National Institutes of Health; Up-Regulation; Virus; cytokine; inhibitor/antagonist; macrophage; neuroinflammation; novel; receptor; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The effects of simian immunodeficiency virus (SIV) on blood-brain barrier permeability are likely initiated by glial response to the presence of virus in the central nervous system. This localized inflammatory reaction to the virus drives dysregulation of the endothelium by RNA modulation and protein phosphorylation. We examined interendothelial signaling by identifying and quantifying SIV-induced pro-inflammatory pathways and resultant message level changes in the endothelium, in response to SIV. We utilized confocal microscopy, cytokine multiplexing, RNA microarray, and RT-PCR. We determined a novel downregulation of CD263, a TNF decoy receptor in SIV stimulated endothelial cells, that normally serves to mitigate the damaging effects of TRAIL when the endothelium encounters an upsurge of proinflammatory stimuli. Supernatant from SIVmac251 infected macrophage was sufficient to cause astrocytes to overproduce TNF-¿ by about 4-fold. Further, the same supernatant increased CCL2 and CXCL8 production in endothelial cells. We used RNA microarray to query RNA regulation in the macaque endothelium caused by SIVmac251 infection. These studies confirm that endothelial inflammation/activation is driven by pro-inflammatory cytokines, especially TNF-¿. The activation of the TNF-¿ receptor in endothelial cells by its ligand causes activation of NF¿B in the endothelium. This leads to the subsequent upregulation/phosphorylation of MLCK, actin cytoskeletal reorganization, and redistribution of ZO-1. Displacement of ZO-1 leads to dysfunction of the endothelial tight junction possibly mediated through JAM-2 and claudin 8. We were able to largely mitigate changes in SIVmac251-driven mRNA changes in primary brain microvascular endothelial cells with inhibitors specific to MLCK and FAK.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 猿猴免疫缺陷病毒 (SIV) 对血脑屏障通透性的影响可能是由神经胶质细胞对中枢神经系统中存在的病毒的反应引发的，这种对病毒的局部炎症反应通过 RNA 调节和蛋白质磷酸化驱动内皮细胞的失调。我们利用共聚焦技术，通过识别和量化 SIV 诱导的促炎症通路以及由此产生的内皮信息水平变化来检查内皮信号传导。我们通过显微镜、细胞因子多重分析、RNA 微阵列和 RT-PCR 确定了 SIV 刺激的内皮细胞中 CD263（一种 TNF 诱饵受体）的新型下调，当内皮遇到促炎刺激激增时，通常可以减轻 TRAIL 的破坏作用。来自 SIVmac251 感染的巨噬细胞的上清液足以导致星形胶质细胞过度产生 TNF-¿此外，相同的上清液使内皮细胞中的 CCL2 和 CXCL8 产量增加了约 4 倍。我们使用 RNA 微阵列来探究 SIVmac251 感染引起的猕猴内皮细胞中的 RNA 调节。炎症细胞因子，尤其是 TNF-¿ .TNF-¿的激活内皮细胞中的受体通过其配体激活 NF¿这导致 MLCK 随后上调/磷酸化、肌动蛋白细胞骨架重组以及 ZO-1 的重新分布。ZO-1 的置换导致内皮连接紧密功能障碍，可能是通过 JAM-2 和密蛋白 8 介导的。我们能够使用 MLCK 和特异性抑制剂很大程度上减轻原代脑微血管内皮细胞中 SIVmac251 驱动的 mRNA 变化。假的。