LEUKOCYTE ADHESION IN ALLERGIC INFLAMMATION

循环中的嗜酸性粒细胞与血管内皮细胞的黏附 炎症组织中血管外部位的积聚是 过敏性炎症。然而，调节的分子机制 体内流动条件下嗜酸性粒细胞黏附情况不佳 明白了。最近对中性粒细胞的研究表明，血管内 这些细胞的黏附是一个多步骤的过程，涉及不同的 粘合和活化步骤。黏附分子包括 选择素家族和β2整合素促进中性粒细胞滚动和 随后的渗出。我们最近观察到，来自 特应性患者类似地滚动并黏附IL-1刺激的内皮细胞 剪切率下的细胞。我们发现一种β1整合素，VLA-4，在 加入L-选择素促进人嗜酸性粒细胞黏附的早期事件 到体内的内皮细胞。拟议研究的目的是 阐明黏附受体在调节人类免疫功能中的作用 体内嗜酸性粒细胞与血管黏附分子的相互作用。vbl.使用 活体显微镜我们将(1)检测L-选择素和VLA-4的作用 整合素促进人嗜酸性粒细胞沿IL-1或IL-4滚动 体内刺激内皮细胞。抗VLA-4和L-选择素的应用 人VLA-4基因的抗体和细胞系 确定两个受体的功能是顺序的还是并行的 促进嗜酸性粒细胞滚动；(2)测定嗜酸性粒细胞活性的作用 细胞因子IL-5和GM-CSF对血管内皮细胞受体激活和体内黏附的影响 循环嗜酸性粒细胞(3)检测趋化因子、RANTES、GM- 脑脊液和纤溶酶原激活剂对受体介导的牢固黏附(粘连)及其后续的影响 正常密度和低密度亚群通过细胞因子的外渗 和(4)确定组胺的作用， GM-CSF、IL-1、IL-4、IL-5对诱导血管表达的影响 内皮细胞表面配体及其促进滚动、粘连的作用 生理状态下活化的嗜酸性粒细胞的跨内皮迁移 体内剪切率。血管粘连的表面表达 在流动条件下，肠系膜中的分子将相互关联。 具有人体嗜酸性粒细胞的体内功能。这些建议的研究 旨在更好地理解分子机制 介导细胞黏附，为检测治疗方案提供依据 过敏性炎症的治疗策略。

Adherence of circulating eosinophils to the vascular endothelium and their accumulation at extravascular sites in inflamed tissues is a hallmark of allergic inflammation. However, the molecular mechanisms mediating eosinophil adhesion under conditions of flow in vivo are not well understood. Recent studies on neutrophils have shown that intravascular adhesion of these cells is a multi-step process involving distinct adhesion and activation steps. Adhesion molecules include members of the selectin family and beta2 integrins which promote neutrophil rolling and subsequent extravasation. We have recently observed that eosinophils from atopic patients similarly roll and adhere to IL-1 stimulated endothelial cells at shear rates. We have found that a beta1 integrin, VLA-4, in addition to L-selectin promotes early events of human eosinophil adhesion to endothelial cells in vivo. The aim of the proposed research is to elucidate the functional role of adhesion receptors in mediating human eosinophil interaction with vascular adhesion molecules in vivo. Using intravital microscopy we will (1) examine the role of L-selectin and VLA-4 integrin in promoting human eosinophil rolling along IL-1 or IL-4 stimulated endothelial cells in vivo. Using anti-VLA-4 and L-selectin antibodies and cell lines transfected with cDNA for human VLA-4 we will determine if both receptors function sequentially or in parallel to promote eosinophil rolling; (2) determine the effects of eosinophil-active cytokines, IL-5 and GM-CSF, on receptor activation and in vivo adhesion of circulating eosinophils (3) examine the effect of chemokines, RANTES, GM- CSF and PAF on receptor mediated firm adhesion (sticking) and subsequent extravasation of normodense and hypodense subpopulations across cytokine stimulated mesenteric EC in vivo and (4) determine the role of histamine, GM-CSF, IL-1, IL-4 and IL-5 on expression of inducible vascular endothelial surface ligands and their role in promoting rolling, sticking and transendothelial migration of activated eosinophils at physiological shear rates in vivo. The surface expression of vascular adhesion molecules in the mesentery under conditions of flow will be correlated with the in vivo function of human eosinophils. These proposed studies are intended to give a better understanding of molecular mechanisms mediating cell adhesion and provide a basis for examining therapeutic strategies for the treatment of allergic inflammation.