CYTOADHESION MOLECULES IN CYTOKINE-DIRECTED EOSINOPOIESIS

建议用实验来检验粘附力的影响。 分子调控的细胞-细胞和细胞-细胞外基质 细胞因子诱导的人祖细胞嗜酸性粒细胞生成的相互作用 细胞。分化抗原表达的研究， CD34和CD33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 使用一种新开发的培养物制剂 来源于人脐血的嗜酸性粒细胞。研究将会 利用多色流式细胞术，祖细胞-ECM 共培养和一种新的内皮细胞黏附试验 评估黏附分子在细胞表面的表达和功能 发育和成熟培养的嗜酸性粒细胞。数据来自完全成熟的、 培养的细胞将与从细胞中提取的细胞进行比较 正常人外周血。中环 第一系列研究的假设是一种特定的CD34 +单个核造血祖细胞亚群 人类脐带血的一部分定义了一个 承诺的前体细胞对造血学有反应 细胞因子、干细胞因子、粒细胞-巨噬细胞集落- 刺激因子(GM-CSF)、白细胞介素3(IL-3)和IL-5。 建议进行实验以分离祖细胞亚群 高纯度的细胞，并在有 不同浓度的单一细胞因子和不同的 细胞因子的组合和序列。研究将揭示 不同细胞的增殖和分化反应 在这些条件下的亚群。我们将并行执行 MRNA表达及颗粒蛋白介体和脂质的研究 由静止和激活的细胞产生的介体 这些研究。第二系列研究将描述 人嗜酸性粒细胞发育过程中表面黏附分子的个体发育 体外及其与细胞外基质分子和/或其他细胞外基质分子的相互作用 脐血单个核细胞组分中存在的细胞。vbl.使用 脐带血系统的嗜酸性粒细胞，特殊的表面分子， ICAM-1、LFA-1、VLA-4和CD44将用单项和多项分析 嗜酸性粒细胞发育过程中的彩色流式细胞术。这些受体 它还可能参与细胞-细胞或细胞外基质-细胞间的相互作用 将在细胞因子的最佳和次优条件下进行研究 刺激。这些研究将提供 细胞因子刺激、黏附分子受体表达和 嗜酸性粒细胞生成。从这些研究中得出的数据应该提供 深入了解黏附分子的双向作用机制 嗜酸性粒细胞的发育和调控，并提示独特 嗜酸性粒细胞增多症的治疗干预途径 哮喘和过敏性疾病中的炎症反应。

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.