EVALUATION OF POLYMORPHONUCLEAR LEUKOCYTE SUBPOPULATIONS

人类多形核白细胞(PMN)亚群已被 基于对包被免疫球蛋白的红细胞的玫瑰花环变化的报道， 趋化作用、FMLP结合和膜电位电荷 刺激。没有注意到它们之间的大小差异 亚群。逆流离心淋洗(CCE)分离细胞 基于大小和密度的差异。该提案的第一阶段将 评估CCE分离的血液PMN的体积依赖亚群。 受体配体结合早期激活事件的分析， 膜电位的变化以及膜结合和胞浆的变化 刺激时将进行钙化。超氧化物已经被 在较大的亚群中表现为在每个细胞的基础上更大。评估 将做为评估PMN亚群滞后时间和氧化率 刺激后爆裂。要确定特定颗粒内容是否具有 在PMN亚群中，Vit B12结合蛋白的作用将被分析。这 分析将评估PMN亚群存在于 处于不同的“激活准备状态”。第二阶段将评估 储存池PMN对PMN亚群的贡献。骨中性粒细胞 骨髓样本将与外周血PMN进行比较。PMN也将被 皮下注射肾上腺素前后分离以评估 PMN的边际池。评价脾在储藏中的作用 PMN和容量依赖的PMN亚群患者接受 择期脾切除术前后将分离中性粒细胞 程序。这些PMN将与外周血PMN进行比较 阶段I中使用的方法分析亚群的迁移 存储池PMN将使用CCE执行。第三阶段将评估 宿主改变患者中性粒细胞亚群的体积依赖关系 防御。Wiskott-Aldrich综合征、慢性肉芽肿性疾病、烧伤 1例PMN乳铁蛋白缺乏症患者和镰状细胞 将对疾病进行评估。我们相信这一信息将有助于 了解中性粒细胞的激活情况。

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.