VASCULAR-MONOCYTE ADHESION MOLECULE IN ATHEROGENESIS

描述（改编自研究者摘要）：本研究的目的 建议是研究单核细胞募集的分子基础， 动脉粥样硬化，重点是分子和细胞生物学的一种新的 单核细胞粘附相关内皮分子151-ELAM。 初步 研究揭示了151-ELAM在单核细胞结合中的主要作用， 单核细胞样细胞系对LPS、IL-1和TNF-α刺激的小鼠 内皮细胞（EC），以最低限度修饰的低密度脂蛋白 刺激兔主动脉EC和完整主动脉段内皮细胞 从胆固醇喂养的兔子在正面结合试验。 我们将探索 151-ELAM表达和调节，确定了其在单核细胞中的重要性 结合在动脉粥样硬化形成，阐明其结构，并询问它是否 代表动脉粥样硬化形成的治疗调节的靶点。 1)的 151-ELAM在小鼠和兔模型中的表达和功能 将探索动脉粥样硬化形成。 分析表达模式， 调节使用常规和正面全主动脉免疫组织化学 方法将是重要的，在了解它的作用，在内皮细胞 功能障碍和单核细胞募集。 的功能特性 与其它已知的粘附分子相比，将研究一种粘附分子， 在单核细胞与来自人的完整主动脉节段结合的离体测定中， 肥肥的兔子 抗151-ELAM治疗对发育的影响 脂饲兔和apoE缺陷小鼠脂肪条纹的形成和发展 将评估模型，以确认其在体内的作用，并评估其 作为治疗靶点的潜力。 2)151-ELAM在 将评估单核细胞-内皮细胞相互作用的多步骤过程 使用基于流动的测定。 循环白细胞与 外渗部位的内皮包括几个连续步骤： 附着、滚动、Galpha-i蛋白连接的激活和 激活依赖性粘附和阻滞。 阻止的步骤 将评估抗151-ELAM单克隆抗体，从而提供关键的见解， 151-ELAM在单核细胞募集中的分子作用。 3)cDNA编码 151 -ELAM将通过标准程序进行克隆和表征。 序列 同源性可以提供对151-ELAM结构和功能的了解。 转染子或重组蛋白将有助于其表征。 粘合性能和特性。 拟议的研究将阐明 这种新型血管受体结构及其在单核细胞中的作用 动脉粥样硬化形成中的募集。 它们可能会导致新的方法， 调节动脉粥样硬化过程。

DESCRIPTION (Adapted from Investigator's Abstract): The objective of this proposal is to study the molecular basis of monocyte recruitment in atherogenesis, focusing on the molecular and cell biology of a novel monocyte adhesion-associated endothelial molecule, 151-ELAM. Preliminary studies reveal a major role for 151-ELAM in binding of monocytes or monocytoid cell lines to LPS, IL-1 and TNF-alpha stimulated mouse endothelial cells (EC), to minimally modified low density lipoprotein stimulated rabbit aortic EC and to endothelium of intact aortic segments from cholesterol fed rabbits in en face binding assays. We shall explore the expression and regulation of 151-ELAM, define its importance in monocyte binding in atherogenesis, elucidate its structure, and ask whether it represents a target for therapeutic modulation of atherogenesis. 1) The expression and function of 151-ELAM in mouse and rabbit models of atherogenesis will be explored. Analysis of the patterns of expression and regulation using conventional and en face whole aorta immunohistologic approaches will be important in understanding its role in endothelial dysfunction and monocyte recruitment. Functional characterization of the molecule, in comparison with other known adhesion molecules, will be pursued in an ex vivo assay of monocyte binding to intact aortic segments from fat-fed rabbits. The effect of anti- 151-ELAM treatment on the development and progression of fatty streaks in fat-fed rabbit and apoE-deficient mouse models will be assessed to confirm its role in vivo and to evaluate its potential as a therapeutic target. 2) The role 151-ELAM plays in the multi-step process of monocyte-endothelial interaction will be assessed using flow based assays. The interaction of circulating leukocytes with endothelium at sites of extravasation involves several sequential steps: attachment, rolling, Galpha-i protein-linked activation and activation-dependent adhesion and arrest. The step or step(s) blocked by anti-151-ELAM MAbs will be assessed, thus providing critical insights into the molecular role of 151-ELAM in monocyte recruitment. 3) cDNAs encoding 151 -ELAM will be cloned and characterized by standard procedures. Sequence homologies may provide insights into 151-ELAM structure and function. Transfectants or recombinant protein will facilitate characterization of its adhesive properties and specificities. The proposed studies will elucidate the structure of this novel vascular receptor and its role in monocyte recruitment in atherogenesis. They may lead to new approaches to the regulation of the atherogenic process.