MECHANISM OF HUMAN TRANSENDOTHELIAL LEUKOCYTE MIGRATION

人类跨内皮白细胞迁移机制

• 批准号: 3082603
• 负责人: SAMUEL CHARLES SILVERSTEIN
• 金额: $ 8.29万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3082603
• 关键词: actins; amnion; calcium; cell migration; cytoskeleton; female; histamine; human fetus tissue; human subject; human tissue; immunochemistry; inflammation; leukotrienes; membrane proteins; myosins; neutrophil; phosphorylation; secretion; serotonin; tissue /cell culture; vascular endothelium permeability

An inflammatory response characterized by leukocyte emigration and increased vascular permeability to fluids and proteins is seen in a number of common disease states including infection, atherosclerosis, acute respiratory distress syndrome, and immune complex diseases. To examine leukocyte migration and alterations in permeability across an endothelial cells (EC) barrier, I have cultured human EC monolayers on a substrate of human amnion. I have measured the permeability of these monolayers and changes in EC cytosolic free calcium ((Ca++)i) during human neutrophil (PMN) migration across such EC monolayers. Transendothelial PMN migration occurs in response to chemoattractants and does not alter the permeability of EC monolayers. Both histamine, which increases EC monolayer permeability, and PMN migration across an EC monolayer induce increases in EC (Ca++)i and changes in EC shape. The goals of the proposed research are: 1) to determine whether PMN surface proteins and/or secretory products initiate EC (Ca++)i changes using antibodies against leukocyte surface proteins and supernatants derived from activated PMN, 2) to examine the role of EC (Ca++)i changes in mediating transendothelial PMN migration or permeability changes by examining the effect of buffering EC (Ca++)i changes on these events, 3) to search for EC myosin light chain phosphorylation or actin rearrangement in stimulated EC as evidence that migrating leukocytes and agonists such as histamine induce EC cytoskeletal changes as a basis for altering EC monolayer permeability, and 4) to examine the relationship between (Ca++)i and changes in the permeability of endothelial and epithelial monolayers by determining if substances which increase the permeability EC monolayers affect EC (Ca++)i and if PMN migration across monolayers of epithelial cells alter epithelial (Ca++)i.

一种以白细胞迁移为特征的炎症反应 可以看到血管对液体和蛋白质的渗透性增加 在包括感染在内的许多常见疾病状态下， 动脉粥样硬化、急性呼吸窘迫综合征和免疫 复杂的疾病。为了检查白细胞的迁移和 内皮细胞(EC)通透性的变化 屏障，我已经在一种基质上培养了人类EC单层 人类羊膜。我测量了这些东西的渗透性 内皮细胞胞浆游离钙((Ca++)i)的单层和变化 在人中性粒细胞(PMN)通过这种EC迁移的过程中 单层。作为回应，发生了跨内皮中性粒细胞迁移 不改变EC的通透性 单层。两种组胺都能增加EC单层 渗透性和PMN在EC单层上的迁移 EC(Ca++)i升高，EC形态发生改变。世界银行的目标是 拟开展的研究工作有：1)确定PMN表面 蛋白质和/或分泌产物启动EC(Ca++)i变化 使用针对白细胞表面蛋白的抗体和 来自活化的PMN的上清液，2)检测其作用 EC(Ca++)i变化在介导跨内皮中性粒细胞变化中的作用 通过检查以下因素的影响来改变迁移或渗透率 缓冲这些事件的EC(Ca++)i变化，3)搜索EC 肌球蛋白轻链磷酸化或肌动蛋白重排 刺激内皮细胞作为迁移白细胞和激动剂的证据 如组胺诱导EC细胞骨架改变作为 改变EC单层渗透性，以及4)检查 (Ca++)i与血管通透性变化的关系 通过确定物质是否为内皮和上皮单分子层 增加通透性的EC单层对EC(Ca++)i的影响 如果PMN跨上皮细胞单层迁移改变 上皮性(Ca++)i。