EOSINOPHIL COLONY FORMING CELLS IN EOSINOPHILIC DISEASES

嗜酸性粒细胞增多症是多种疾病状态的特征， 哮喘和特应性皮炎，寄生虫感染和 嗜酸性粒细胞增多综合征（HES）。 影响干细胞的因素 嗜酸性粒细胞线的承诺和随后的过度 这些疾病中的嗜酸性粒细胞生成在很大程度上未被探索。 嗜酸性 集落形成细胞（EO-CFC）是主要的集落形成细胞， 外周血正常，但循环嗜酸性粒细胞仅占1-3% 正常循环中白细胞的数量。 其意义 差异和循环EO-CFC在组织生产中的作用 嗜酸性粒细胞增多症尚未确定。 HES患者具有异质性 根据他们的临床表现，血液嗜酸性粒细胞增多症的水平， 他们的预测。 在我们提出的研究中，我们将定量骨中的EO-CFC 骨髓和外周血HES患者，并检测其集落形成 对外源性和自体来源的 集落刺激因子（CSF）。 对普通CFC的实验将在 以确定HES患者是否产生过量 大量的EO-CSF。 HES患者的EO-CFC 将通过检查它们的生长情况来测试半自主增殖 在次优培养条件下的潜力， 当注射到裸鼠中时，再现HES的特征。 接下来我们就 使用低密度细胞组分生产抗EO-CFC的单克隆抗体 从在软琼脂或甲基纤维素培养物中生长的嗜酸性粒细胞菌落， 免疫原 将筛选抗-EO-CFC单克隆抗体的反应性 针对成熟嗜酸性粒细胞、多形核细胞、成纤维细胞和 粒细胞-巨噬细胞CFC。 抗体从细胞中选择EO-CFC的能力 将利用荧光测定混合细胞群 标记和未标记的活化细胞分选和软琼脂培养 细胞 使用荧光抗体技术，单克隆抗体将 用作探针检测组织中EO-CFC的存在， 通常含有大量的嗜酸性粒细胞。 最后，我们将测试 单克隆抗体单独和与蓖麻毒蛋白偶联时 链-A在体外选择性杀死正常和HES EO-CFC。

Eosinophilia is characteristic of a variety of disease states ranging from asthma and atopic dermatitis to parasitic infestations and the hypereosinophilic syndrome (HES). The factors influencing stem cell commitment to the eosinophil line and subsequent excessive eosinophilopoiesis in these diseases are largely unexplored. Eosinophil colony-forming cells (EO-CFC) are the predominant colony-forming cell in normal peripheral blood, yet, circulating eosinophilis constitute only 1-3% of the normal circulating leukocyte population. The significance of this discrepancy and the role of circulating EO-CFC in production of tissue eosinophilia have not been determined. Patients with HES are heterogeneous in terms of their clinical presentations, level of blood eosinophilia and their prognosis. In our proposed studies we will quantitate EO-CFC in bone marrow and peripheral blood patients with HES and test their colony-forming ability in response to exogenous and autologous sources of colony-stimulating factor (CSF). Experiments on normal CFC will be conducted to determine whether patients with HES produce excessive quantities of EO-CSF. The ability of EO-CFC from HES patients to proliferate semiautonomously will be tested by examination of their growth potential under suboptimal culture conditions and by their ability to reproduce features of HES when injected into nude mice. Next, we will produce monoclonal antibody to EO-CFC using light density cell fractions from eosinophil colonies grown in soft agar or mythylcellulose culture as the immunogen. Monoclonal anti-EO-CFC will be screened for its reactivity against mature eosinophils, polymorphonuclear cells, fibroblasts and granulocyte-macrophage CFC. The antibody's ability to select EO-CFC from a mixed population of cells will be determined utilizing fluorescence activated cell sorting and soft agar culture of labeled and unlabeled cells. Using a fluorescent antibody technique the monoclonal antibody will be used as a probe to detect the presence of EO-CFC in tissues which normally contain large numbers of eosinophils. Lastly, we will test the ability of the monoclonal antibody both alone and when conjugated to ricin chain-A to selectively kill normal and HES EO-CFC in vitro.