EOSINOPHIL COLONY FORMING CELLS IN EOSINOPHILIC DISEASES

嗜酸性粒细胞增多症是各种疾病状态的特征，范围从 哮喘和特应性皮炎与寄生虫感染和 高嗜酸性粒细胞综合征(HES)。影响干细胞的因素 对嗜酸性粒细胞路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是未知的。嗜酸性粒细胞 集落形成细胞(EO-cfc)是最主要的集落形成细胞。 外周血正常，但循环中的嗜酸性粒细胞仅占1-3% 正常循环中的白细胞数量。这件事的意义 循环EO-CFC在纸巾生产中的差异和作用 嗜酸性粒细胞增多症尚未确定。患有HES的患者是不同的 根据他们的临床表现，血嗜酸粒细胞水平和 他们的预后。在我们拟议的研究中，我们将对骨骼中的EO-CFC进行量化 HES患者骨髓和外周血细胞集落形成的检测 对外源和自体来源的反应能力 集落刺激因子(CSF)。普通氯氟化碳的实验将 以确定患有HES的患者是否产生过量的 EO-CSF的数量。HES患者外周血中EO-CFCs的能力 将通过检查它们的生长情况来测试半自主增殖 在不理想的培养条件下的潜力和他们的能力 将HES注射到裸鼠体内，复制HES的特征。接下来，我们将 利用光密度细胞组分制备抗EO-CFCs的单抗 从软琼脂或神话纤维培养物中培养的嗜酸性粒细胞集落 免疫原。将对抗EO-CFC的单抗进行反应性筛选 抗成熟的嗜酸性粒细胞、多形核细胞、成纤维细胞和 粒细胞-巨噬细胞CFs。抗体选择EO-CFC的能力 混合的细胞群体将利用荧光来确定 标记和非标记细胞的活化分选和软琼脂培养 细胞。使用荧光抗体技术，单抗将 被用作探针来检测组织中是否存在EO-CFC， 通常含有大量的嗜酸性粒细胞。最后，我们将测试 单抗单独和与蓖麻毒素结合的能力 链-A在体外选择性地杀死正常和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.