MONONUCLEAR LEUKOCYTE ADHESION AND RECRUITMENT IN CHRONIC INFLAMMATORY DISEASE

选择素、整合素和免疫球蛋白基因的黏附受体 家庭被认为是正常和病态的 所有白细胞的招募；然而，大多数研究都集中在 中性粒细胞和急性炎症性损伤。粘连堵塞 参与募集的受体限制中性粒细胞依赖的损伤 一些模型系统，特别是在肺部。这些成功 带来希望，类似的方法将有利于慢性炎症性和 免疫介导的疾病也是如此。然而，黏附受体可能会 参与招募单核白细胞，关键 这些疾病的细胞介体还没有得到充分的评价 在生理相关的模型系统中。从中性粒细胞中可以明显看出 必须在剪切条件下研究白细胞-内皮细胞黏附的研究- 压力决定单个黏附分子的贡献 到活体内招募。同样重要的是评估 受体阻断对相关基因募集影响的定量研究 慢性炎症性疾病的模型，因为体外建模，充其量， 接近活体内的流动条件。这样的研究是有基础的 发展以招聘为目标的治疗学的重要性 慢性炎症性/免疫性疾病。我们的初步研究 确定含有α4的整合素，很可能是α4/β1，作为关键 单核白细胞与细胞因子处理后黏附的受体 剪切力作用下的内皮细胞。取决于其基本的绑定水平 活性，这种受体要么加强E-启动的依恋 选择或启动附加和招聘。前者 主要用于循环细胞共表达的对抗性受体 选择素和低亲和力的α4整合素。后者 主要用于循环中的“激活”单核细胞和淋巴母细胞 在抗原性或炎症性刺激下产生的。在……里面 特异性靶向1(SA 1)，阻断特异性的单抗 β1链和/或“高亲和力”形式的表位 Alpha4/Beta1受体将在三个物种中产生。这些单抗 将对SAS 204做出贡献。SA 2将探索E-的相互作用 黏附过程中的选择素、L-选择素与α4/β1受体 纯化的人单眼白细胞亚群和相关细胞系 细胞因子激活的内皮细胞在生理水平的线性剪切- 体外应激。SA 3将探索细胞激活在 剪切力的上调与激活的表达 α/β受体上的表位。SA 4将使用新开发的 和目前可用的针对α4/β1受体的单抗 确定该受体对单核白细胞的贡献 葡聚糖诱导的大鼠肺肉芽肿中的募集(协作 与J.Warren，项目III和P.A.Ward，项目I)和血吸虫- 诱导的小鼠肺肉芽肿(与S.kunkel合作， 项目II)。

Adhesion receptors from the selectin, integrin and immunoglobulin gene families are thought to be instrumental in the normal and pathologic recruitment of all leukocytes; however, most studies have focused on neutrophils and acute inflammatory injury. Blockage of adhesion receptors involved in recruitment limits neutrophil-dependent injury in a number of model systems particularly in the lung. These successes raise hope that a similar approach will benefit chronic inflammatory and immune mediated diseases as well. However, adhesion receptors likely to be involved in the recruitment of mononuclear leukocytes, the key cellular mediators of these diseases, have not been adequately evaluated in physiologically relevant model systems. It is clear from neutrophil studies that leukocyte-endothelial adhesion must be studied under shear- stress to determine the contributions of individual adhesion molecules to recruitment in vivo. It is equally important to evaluate quantitatively the impact of receptor blockade on recruitment in relevant models of chronic inflammatory disease since in vitro modeling, at best, approximates flow conditions in vivo. Such studies are of fundamental importance to the development of therapeutics targeted at recruitment in chronic inflammatory/immunologic diseases. Our preliminary studies identify alpha4-containing integrins, most likely alpha4/Beta1, as a key receptor in the attachment of mononuclear leukocytes to cytokine-treated endothelium under shear. Depending on its basal level of binding activity, this receptor either strengthens attachment initiated by E- selectin or initiates attachment and recruitment. The former predominates for circulating cells co-expressing counter-receptors for the selectins and "low avidity" alpha4 integrins. The latter predominates for circulating "activated" monocytes and lymphoblasts generated in response to antigenic or inflammatory stimulate. In Specific Aim 1 (SA 1), blocking monoclonal antibodies (mabs) specific for epitopes on the putative "high avidity" form of the Beta1 chain and/or the alpha4/beta1 receptor will be generated in three species. These mabs will contribute to SAs 204. SA 2 will explore the interactions of E- Selectin, L-Selectin and the alpha4/Beta1 receptor during attachment of purified subsets of human monocular leukocytes and related cell lines to cytokine-activated endothelium at physiologic levels of linear shear- stress in vitro. SA 3 will explore the role of cellular activation in upregulating the function under shear and expression of activation epitopes on the alpha/beta receptor. SA 4 will use the newly developed and currently available Mabs specific for alpha4/beta1 receptor to determine the contribution of this receptor to mononuclear leukocyte recruitment in glycan-induced lung granulomas in the rat (collaboration with J. Warren, project III and P.A. Ward, project I) and schistosome- induced lung granulomas in the mouse (collaboration with S. Kunkel, project II).