CELLULAR ATTACHMENT AND ADHESIVE RECEPTORS

单核细胞在血管壁上的附着、黏附和渗透 代表了一系列血管疾病的特征，包括 血栓形成、动脉粥样硬化和炎症。内皮细胞表面 表达附着分子，如E-和P-选择素，以及 黏附分子，如ICAM-1和VCAM-1，局部诱导 炎症介质在单核细胞的调节中起着重要作用 通过暴露特定反物的结合部位进行附着和粘连 单核细胞表面的受体。这项提议的中心前提是 单核细胞上特定的附着受体的结合是一种 单核细胞活化的重要途径及其依恋诱导 激活触发一条独特的细胞内信号通路，该通路具有 尤其与动脉粥样硬化的发生有关。已获得初步数据 表明当外周血单核细胞与 细胞因子激活的内皮细胞一种特殊的表型变化 在单核细胞中诱导，包括促凝血剂的表面表达 组织因子，CD36(一种糖蛋白)表面表达增加 清道夫受体和黏附受体)，以及分泌前列环素 炎性细胞因子肿瘤坏死因子。这种表型变化是 特定基因转录增加，需要直接接触 在内皮细胞和单核细胞之间。E-选择素的参与 单核细胞上的受体已被证明在诱导 这些表型变化。勾勒出定义表面的平面图， 单核细胞参与的细胞质和核信号通路 由细胞附着受体激活。尤其是电子邮件的作用 选择素配体(ES-1和CD15)将用免疫抑制法进行探索 和反义寡核苷酸方法。单核细胞与E-的黏附 选择素转染细胞将用于探索特定的 胞质内信号通路与核内基因调控 E-选择素参与的机制。这些研究将涉及 与D.Hajjar博士和B.Hempstead博士密切互动，他们是 细胞信号通路。由该途径激活的特定基因将 经Northern分析、聚合酶链式反应和酶联免疫吸附试验鉴定 关注与血管相关的单核/巨噬细胞效应功能 生物学。这将涉及到与D.Hajjar博士和A. 马库斯正在研究清道夫受体、细胞因子和二十烷类化合物； K.Hajjar正在研究凝血和纤溶的调节剂。小说 E-选择素激活的基因将通过聚合酶链式反应进行鉴定 差异显示和/或正选择克隆技术。它是 期望拟议的研究可以提供新的机制洞察力 变成细胞信号，并最终可能允许开发新的 血管和炎症性疾病的治疗策略。

Monocyte attachment, adhesion, and infiltration into the vessel wall represent hallmarks of a wide range of vascular disorders including thrombosis, atherosclerosis, and inflammation. Endothelial cell surface expression of attachment molecules, such as E- and P-selectin, and adhesion molecules, such as ICAM-1 and VCAM-1, locally induced by inflammatory mediators, plays an important role in mediating monocyte attachment and adhesion by exposing binding sites for specific counter- receptors on the monocyte surface. The central premise of this proposal is that engagement of specific attachment receptors on monocytes is an important pathway of monocyte activation, and that attachment-induced activation triggers a unique intracellular signalling pathway that has particular relevance to atherogenesis. Preliminary data has been obtained demonstrating that when peripheral blood monocytes are co-cultured with cytokine-activated endothelial cells a specific phenotypic change is induced in the monocytes that includes surface expression of procoagulant tissue factor, increased surface expression of CD36 (a glycoprotein scavenger receptor and adhesion receptor), and secretion of the pro- inflammatory cytokine TNF. This phenotypic change is the result of increased transcription of specific genes and requires direct contact between the endothelial cells and the monocytes. Engagement of E-selectin receptors on monocytes has been shown to play a major role in inducing these phenotypic changes. Plans are outlined to define the surface, cytoplasmic and nuclear signalling pathways involved in monocyte activation by cellular attachment receptors. In particular the role of E- selectin ligands (ES-1 and CD15) will be explored using immuno-inhibition and anti-sense oligonucleotide approaches. Attachment of monocytes to E- selectin transfected cells will be used to explore specific intracytoplasmic signalling pathways and intranuclear gene regulation mechanisms induced by E-selectin engagement. These studies will involve close interactions with Dr. D. Hajjar and B. Hempstead who are experts in cell signalling pathways. Specific genes activated by this pathway will be characterized by northern analysis, PCR, and ELISA with particular attention to monocyte/macrophage effector functions relevant to vascular biology. This will involve close interactions with Dr. D. Hajjar and A. Marcus looking at scavenger receptors, cytokines, and eicosanoids; and Dr. K. Hajjar looking at regulators of coagulation and fibrinolysis. Novel genes activated by E-selectin engagement will be identified by PCR differential display and/or positive selection cloning techniques. It is expected that the proposed studies may provide novel mechanistic insight into cell signalling and ultimately may allow development of novel therapeutic strategies for vascular and inflammatory diseases.