Leukotrienes and blood-brain barrier

描述(申请人提供)：微生物穿越血脑屏障是发展中枢神经系统感染所必需的，但其潜在机制仍不完全清楚。血脑屏障是由脑微血管内皮细胞形成的结构和功能屏障，保护大脑免受血液中循环的任何微生物的影响，但最近的研究表明，引起脑膜炎的病原体能够作为活的生物体穿过血脑屏障。我们通过分离和培养人脑微血管内皮细胞(HBMEC)建立了体外血脑屏障模型。在胶原涂层的Transwell植入物上培养后，HBMEC单层显示出紧密连接形成和极性单层的形态和功能特性。我们的初步研究表明，引起脑膜炎的病原体穿过HBMEC单层而不影响HBMEC的完整性，通过跨内皮细胞的电阻和通透性来评估，微生物穿越HBMEC单层涉及特定的宿主信号转导途径。我们使用药物抑制和基因缺失进行的其他初步研究首次表明，宿主5-脂氧合酶可能参与微生物穿越血脑屏障。白三烯是由5-脂氧合酶合成的，在呼吸系统疾病、变态反应性疾病和心血管疾病中具有病理生理作用，但它们在微生物穿越血脑屏障中的作用尚未被探索。我们对受体拮抗剂的初步研究表明，半胱氨酰白三烯可能参与微生物穿越血脑屏障的过程。这项应用的总体目标是调查和表征参与微生物穿越血脑屏障的宿主信号分子。从这项应用中获得的信息将展示5-脂氧合酶和白三烯对微生物穿越血脑屏障的新贡献。

DESCRIPTION (provided by applicant): Microbial traversal of the blood-brain barrier is required for the development of central nervous system infection, but the underlying mechanisms remain incompletely understood. The blood-brain barrier is a structural and functional barrier that is formed by brain microvascular endothelial cells and protects the brain from any microorganisms circulating in the blood, but recent studies have shown that meningitis-causing pathogens are able to cross the blood-brain barrier as live organisms. We have developed the in vitro blood-brain barrier model by isolation and cultivation of human brain microvascular endothelial cells (HBMEC). Upon cultivation on collagen-coated Transwell inserts the HBMEC monolayers exhibit morphological and functional properties of tight junction formation and polar monolayer. Our Preliminary Studies revealed that meningitis-causing pathogens traverse the HBMEC monolayers without affecting the HBMEC integrity, as assessed by transendothelial electrical resistance and permeability, and microbial traversal of the HBMEC monolayers involves specific host signal transduction pathways. Our additional Preliminary Studies using pharmacological inhibition and gene deletion suggest for the first time that host 5-lipoxygenase is likely to be involved in microbial traversal of the blood-brain barrier. Leukotrienes are synthesized by 5-lipoxygenase and shown to have pathophysiological roles in respiratory diseases, allergic diseases and cardiovascular diseases, but their role in microbial traversal of the blood-brain barrier has not been explored. Our Preliminary Studies with the receptor antagonists revealed that cysteinyl leukotrienes are likely to be involved in microbial traversal of the blood-brain barrier. The overall aim of this application is to investigate and characterize the host signaling molecules involved in microbial traversal of the blood- brain barrier. The information derived from this application will demonstrate the novel contributions of 5- lipoxygenase and leukotrienes to microbial traversal of the blood-brain barrier.