EVALUATION OF POLYMORPHONUCLEAR LEUKOCYTE SUBPOPULATIONS

人类多形核白细胞（PMN）的亚群已被证实是 根据对IgG包被的红细胞的玫瑰花结的变化报道， 趋化性、FMLP结合和膜电位电荷 刺激. 在这些之间没有注意到尺寸差异 亚群 逆流离心淘洗（CCE）分离细胞 基于大小和密度的差异。 该提案的第一阶段将 评价通过CCE分离的血液PMN的体积依赖性亚群。 受体配体结合的早期活化事件的分析， 膜电位变化，以及膜结合和细胞溶质 将进行刺激后的钙。 超氧化物已经 显示出在较大的子组中每个细胞的基础上更大。 评价 将进行评估PMN亚群的滞后时间和氧化速率， 在刺激后爆裂。 为了确定特定的颗粒含量是否具有 分析Vit B12结合蛋白在PMN亚群中的作用。 这 分析将评估PMN亚群存在于 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PMN 将骨髓样本与外周血PMN进行比较。 PMN也将 在皮下注射肾上腺素之前和之后分离，以评估 PMN的边缘池。 评价脾脏在贮藏中的作用 PMN和容量依赖性PMN亚群患者接受 选择性脾切除术将在此之前和之后分离PMN procedure. 将这些PMN与外周血PMN进行比较， 第一阶段使用的方法。 人口亚群变化分析 将使用CCE执行存储池PMN。 第三阶段将评估 宿主改变患者的体积依赖性PMN亚群 的防御合作. Wiskott-Aldrich综合征，慢性肉芽肿病，烧伤 患者，PMN乳铁蛋白缺乏症患者，镰状细胞 将对疾病进行评估。 我们认为，这些信息将有助于 了解PMN激活。

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.