CELLULAR ATTACHMENT AND ADHESIVE RECEPTORS

单核细胞附着、粘附和浸润血管壁 代表了多种血管疾病的特征，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，和 粘附分子，如ICAM-1和VCAM-1，局部诱导 炎症介质，在介导单核细胞 附着和粘附通过暴露特定的反- 单核细胞表面的受体。 这项提议的核心前提是 单核细胞上特异性附着受体的参与是 单核细胞活化重要途径， 激活触发了一个独特的细胞内信号通路， 特别是与动脉粥样硬化的关系。 初步数据已经获得 这表明当外周血单核细胞与 精氨酸激活的内皮细胞的一种特异性表型变化是 在单核细胞中诱导，包括促凝血剂的表面表达 组织因子，CD 36（一种糖蛋白）表面表达增加 清道夫受体和粘附受体），并分泌前- 炎性细胞因子TNF。 这种表型变化是 增加特定基因的转录，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显示在诱导 这些表型变化。 计划概述，以确定表面， 单核细胞胞质和核信号通路 通过细胞附着受体激活。 尤其是E- 选择素配体（ES-1和CD 15）将使用免疫抑制来探索 和反义寡核苷酸方法。 单核细胞与E- 选择素转染的细胞将用于探索特异性的 胞质内信号通路与核内基因调控 E-选择素参与诱导的机制。 这些研究将涉及 与D博士的亲密接触哈贾和B。亨普斯特德是 细胞信号通路。 通过这种途径激活的特定基因将 通过北方分析、PCR和ELISA进行表征， 关注与血管相关的单核细胞/巨噬细胞效应子功能 生物学 这将涉及与D博士的密切互动。Hajjar和A. 马库斯研究了清道夫受体、细胞因子和类花生酸; K. Hajjar研究凝血和纤维蛋白溶解的调节因子。 小说 将通过PCR鉴定由E-选择素接合激活的基因 差异显示和/或阳性选择克隆技术。 是 预计拟议的研究可能会提供新的机制见解 转化为细胞信号，最终可能允许开发新的 血管和炎性疾病的治疗策略。

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.