CYTOADHESION MOLECULES IN CYTOKINE-DIRECTED EOSINOPOIESIS

实验提出了检查的影响，粘附 分子调控的细胞-细胞和细胞-细胞外基质 人祖细胞中精氨酸介导的嗜酸性粒细胞生成的相互作用 细胞分化抗原表达的研究， CD 34和CD 33以及编码嗜酸性粒细胞主要碱性蛋白的mRNA (MBP)和嗜酸性粒细胞过氧化物酶（EPO）在嗜酸性粒细胞生成过程中， 使用新开发的培养的 来源于人脐带血的嗜酸性粒细胞。研究将 使用多色流式细胞术、祖细胞-ECM 共培养和新开发的内皮细胞粘附试验， 评估粘附分子的表达和功能， 发育和成熟的培养嗜酸性粒细胞。数据来自完全成熟的， 将培养的细胞与从细胞中提取的细胞进行比较。 正常志愿者的外周血。中央 第一系列研究的假设是， + 单核细胞中存在的祖细胞亚群 人脐带血的一部分定义了一个群体， 定型前体细胞对造血细胞的增殖 细胞因子、干细胞因子（SCF）、粒细胞-巨噬细胞集落- 刺激因子（GM-CSF）、白细胞介素（IL）-3和IL-5。 实验提出了分离祖细胞亚群 高纯度的细胞，并在存在下培养这些细胞 不同浓度的单一细胞因子和各种 细胞因子的组合和序列。 研究将揭示 各种细胞的增殖和分化反应 在这些条件下的亚群。我们将进行平行 mRNA表达和颗粒蛋白介质和脂质的研究 由静止和活化细胞产生的介质产生， 这些研究。第二系列研究将描述 嗜酸性粒细胞表面粘附分子的个体发生 体外和它们与ECM分子和/或与其它 存在于脐带血单核细胞部分中的细胞。使用 脐血系统的嗜酸性粒细胞生成，特定的表面分子， ICAM-1、LFA-1、VLA-4和CD 44将通过单细胞和多细胞分析。 在嗜酸性粒细胞的发展过程中的彩色流式细胞术。这些受体 其可能参与细胞-细胞或ECM-细胞相互作用， 将在细胞因子的最佳和次佳条件下进行研究 刺激.这些研究将提供直接的相关性， 精氨酸刺激，粘附分子受体表达和 嗜酸性粒细胞生成从这些研究中获得的数据应提供 粘附分子双向作用机制的研究进展 的发展和调节，并提出独特的 嗜酸性粒细胞增多症的治疗干预途径 在哮喘和过敏性疾病中观察到的炎症反应。

Experiments are proposed to examine the effects of adhesion molecule-regulated cell-cell and cell-extracellular matrix (ECM) interactions on cytokine-directed eosinopoiesis of human progenitor cells. Studies of the expression of the differentiation antigens, CD34 and CD33, and mRNA encoding eosinophil major basic protein (MBP) and eosinophil peroxidase (EPO) during eosinopoiesis will be performed using a newly developed preparation of cultured eosinophils derived from human umbilical cord blood. Studies will be performed utilizing multi-color flow cytometry, progenitor-ECM coculture and a newly developed endothelial cell adhesion assay to assess the expression and functionality of adhesion molecules on developing and mature cultured eosinophils. Data from fully mature, cultured cells will be compared to cells extracted from the peripheral blood from normal human volunteers. The central hypothesis of the first series of studies is that a specific CD34 + subpopulation of progenitor cells present in the mononuclear fraction of human umbilical cord blood defines a population of committed precursor cells responsive to the hematopoietic cytokines, stem cell factor (SCF), granulocyte-macrophage colony- stimulating factor (GM-CSF), interleukin (IL)-3 and IL-5. Experiments are proposed to isolate subpopulations of progenitor cells in high purity and to culture these cells in the presence of different concentrations of single cytokines and in various combinations and sequences of cytokines. Studies will reveal proliferative and differentiative responses of the various subpopulations under these conditions. We will perform parallel studies of mRNA expression and granule protein mediator and lipid mediator production by quiescent and activated cells produced in these studies. A second series of studies will characterize the surface adhesion molecule ontogeny of developing eosinophils in vitro and their interactions with ECM molecules and/or with other cells present in the cord blood mononuclear cell fraction. Using the cord blood system of eosinopoiesis, specific surface molecules, ICAM-1 LFA-1, VLA-4 and CD44 will be analyzed by single and multi- color flow cytometry during eosinophil development. These receptors which may be involved in cell-cell or ECM-cell interactions also will be studies under optimal and suboptimal conditions of cytokine stimulation. These studies will provide direct correlations between cytokine-stimulation, adhesion molecule receptor expression and eosinopoiesis. Data derived from these studies should provide insight into the bi-directional mechanisms of adhesion molecule development and regulation in eosinopoiesis and suggest unique avenues of therapeutic intervention in the eosinophilic inflammatory responses seen in asthma and allergic disease.