EARLY HEMATOPOIETIC CELL REGULATORS OF HUMAN ORIGIN

本项目的目的是研究早期造血的作用。 细胞调节因子，如促进红细胞爆裂的活性 (双酚A)巨核细胞集落刺激因子与集落 形成单位脾(CFU-S)生存促进因子(CFU-S-F)的作用 关于造血细胞的复制、承诺和分化 祖细胞。它意在追求以下目标：孤立， 逐步提纯和鉴定人类来源的相关因子 尿液和人单核细胞条件培养液的技术 包括粒度选择性超滤，分级沉淀， 层析和超速离心法。研究黄连的作用机理 这些因素，例如关于可逆性、时间进程、剂量 靶细胞预孵育过程中的依赖性 含促红细胞生成素(EPO)刺激的悬浮培养 在半固体培养中的细胞或对其进行细胞分析 CFU-S的内容。确定红系数量的增加是否 冲击地压形成单位(BFU-E)还是CFU-S所带来的这些因素 基于细胞复制或细胞激活(敏化)。 调查因素的目标细胞的性质(直接与 间接作用，单能与多能祖细胞)。研究一下 各因素在不同阶段的特殊性 净化。研究指导制定《公约》的机制 并确定其生理和临床意义。至 这一末端细胞群体将被分离，并对分离株进行 在体外进行适当的刺激。双酚A、脑脊液、粒-巨噬细胞集落形成单位S-F含量 贫血、红细胞增多症和血小板紊乱患者的尿液 从不同的原因和正常的个人将被测量以检测 可能与其他血液学参数相关。鲜血 患者的单个核细胞将被测试产生双酚A的能力 在PHA治疗后，也用于对BPA的反应能力 直接和预孵化BFU-E检测，以了解 这些测量提供了临床上有用的信息。基于 在这个项目的过程中获得的信息分析了这些因素 将进一步发展。

The objective of this project is to study the roles early hematopoietic cell regulatory factors such as erythroid burst promoting activities (BPA's) megakaryocyte colony stimulating factor (MEF-CSF) and colony forming unit spleen (CFU-S) survival enhancing factor (CFU-S-F) play with respect to the replication, commitment and differentiation of hematopoietic progenitor cells. It is intended to pursue the following aims: Isolate, progressively purify and characterize relevant factors derived from human urine and from human mononuclear cell conditioned media by techniques involving size selective ultrafiltration, fractional precipitation, chromatography and ultracentrifugation. Study the mechanism of action of these factors, e.g. with respect to reversibility, time course, dose dependence by procedures involving preincubation of the target cells in suspension culture with a factor followed by erythropoietin (EPO) challenge of the cells in semi-solid medium culture or assay of the cells for their content of CFU-S. Determine whether the increase in numbers of erythroid burst forming units (BFU-E) or CFU-S brought about by these factors is based on cell replication or on cell activation (sensitization). Investigate the nature of the target cells for the factors (direct vs. indirect action, unipotent vs. multipotent progenitor cells). Study the specificity of the factors of the factors, at various stages of purification. Study the mechanisms governing the elaboration of the factors and ascertain their physiological and clinical significance. To this end cell populations will be fractionated and the isolates subjected to appropriate stimuli in vitro. The BPA, MEG-CSF and CFU-S-F content of urine from patients with anemias, polycythemias and platelet disorders due to different causes and from normal individuals will be measured to detect possible correlations with other hematological parameters. The blood mononuclear cells of patients will be tested for the ability to produce BPA after PHA treatment and also for the ability to respond to BPA in the direct and the preincubation BFU-E assays in order to find out whether these measurements provide clinically useful information. Based on information gained in the course of this project assays for the factors will be further developed.