EVALUATION OF POLYMORPHONUCLEAR LEUKOCYTE SUBPOPULATIONS

多形核白细胞亚群的评估

• 批准号: 3445645
• 负责人: ROGER L BERKOW
• 金额: $ 5.02万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-30 至 1987-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3445645
• 关键词: Wiskott Aldrich syndrome; blood cell count; bone marrow; burns; cell membrane; cell morphology; cell population study; chronic granulomatous disease; flow cytometry; fluorescent dye /probe; human tissue; immune adherence reaction; immunodeficiency; leukocyte activation /transformation; leukocyte activation disorder; membrane activity; membrane potentials; neutrophil; peripheral blood vessel; sickle cell anemia; splenectomy; superoxides; vascular endothelium

Subpopulations of human polymorphonuclear leukocytes (PMN) have been reported based on variations in rosetting to IgG coated erythrocytes, chemotaxis, binding of FMLP and membrane potential charge upon stimulation. No size differences have been noted among these subpopulations. Counterflow centrifugal elutriation (CCE) separates cells based on differences in size and density. Phase I of the proposal will evaluate volume dependent subpopulations of blood PMN isolated by CCE. Analysis of the early activation events of receptor ligand binding, membrane potential change, and alteration in membrane bound and cytosolic calcium upon stimulation will be performed. Superoxide has already been shown to be greater on a per cell basis in larger subgroups. Evaluation will be done to assess PMN subpopulation lag time and rate of the oxidative burst after stimulation. To determine if specific granule contents have a role in PMN subpopulations, Vit B12 binding protein will be analyzed. This analysis will assess the hypothesis that subpopulations of PMN exist in varying states of "readiness for activation". Phase II will assess the contribution of storage pool PMN to PMN subpopulations. PMN from bone marrow samples will be compared to peripheral blood PMN. PMN will also be isolated before and after a subcutaneous injection of epinephrine to assess the marginating pool of PMN. To evaluate the role of the spleen in storage of PMN and in volume dependent PMN subpopulations patients undergoing elective splenectomy will have PMN isolated before and after this procedure. These PMN will be compared to peripheral blood PMN by the methods utilized in Phase I. Analysis of shifts in subpopulations of the storage pool PMN will be performed utilizing CCE. Phase III will evaluate volume dependent PMN subpopulations of patients with altered host defenses. Wiskott-Aldrich syndrome, chronic granulomatous disease, burn patients, a patient with PMN lactoferrin deficiency, and sickle cell disease will be assessed. We believe that this information will aid in the understanding of PMN activation.

人类多形核白细胞 (PMN) 亚群已被 根据 IgG 包被红细胞玫瑰花结的变化进行报告， 趋化性、FMLP 的结合和膜电位电荷 刺激。 这些之间没有注意到尺寸差异 亚人群。 逆流离心淘洗 (CCE) 分离细胞 基于尺寸和密度的差异。 该提案的第一阶段将 评估通过 CCE 分离的血液 PMN 的体积依赖性亚群。 分析受体配体结合的早期激活事件， 膜电位变化以及膜结合和胞质的改变 将在刺激后进行钙治疗。 超氧化物已经 在较大的亚组中，以每个细胞为基础显示出更大的值。 评估 将评估 PMN 亚群的滞后时间和氧化速率 刺激后爆发。 确定特定颗粒含量是否具有 为了了解 Vit B12 结合蛋白在 PMN 亚群中的作用，我们将进行分析。 这 分析将评估以下假设：PMN 亚群存在于 “准备激活”的不同状态。 第二阶段将评估 存储池 PMN 对 PMN 子群的贡献。 来自骨骼的中性粒细胞 骨髓样本将与外周血 PMN 进行比较。 PMN 也将 在皮下注射肾上腺素之前和之后进行分离以评估 PMN 的保证金池。 评估脾脏的储存作用 PMN 和体积依赖性 PMN 亚群患者接受 选择性脾切除术在此之前和之后将分离 PMN 程序。 这些 PMN 将与外周血 PMN 进行比较 第一阶段中使用的方法。亚群变化分析 存储池 PMN 将利用 CCE 执行。 第三阶段将评估 宿主改变患者的体积依赖性中性粒细胞亚群 防御。 Wiskott-Aldrich 综合征、慢性肉芽肿病、烧伤 患者，患有 PMN 乳铁蛋白缺乏症和镰状细胞病的患者 将评估疾病。 我们相信这些信息将有助于 了解 PMN 激活。