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Purification and characterization of burst-promoting activity

爆发促进活性的纯化和表征

基本信息

批准号：
61560094
负责人：
RYUZO Sasaki
金额：
$ 1.22万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

Two distinct growth factors are needed for erythropoiesis. Erythropoietin (Ep) promotes both the growth of late erythroid precursor cells (CFU-E) and their maturation into proerythroblasts in which hemoglobin (Hb) synthesis is initiated. The other factor, burst-promoting activity (BPA) acts on the early precursor cells (BFU-E) and is responsible for differentiation of these cells into CFU-E with high responsiveness to Ep. This project was aimed at purifying BPA from a human source and exploring the effects of the two factors, BPA and Ep, on erythroid burst-colony formation of human peripheral blood mononuclear cells in methlycellulose cultures. Ep was isolated from the urine of patients with aplastic anemia and BPA was found in the resiue obtained after removal of Ep. Extensive purification of BPA yielded a preparation of BPA completely free from Ep. Reddish bursts were formed with the addition of Ep alone. Addition of BPA not only elevated the number of bursts but also grealty reduced the amount of Ep reqiured for burst formation. The presence of BPA alone in cultures did not permit bursts to form but did permit the growth of small colonies that did not contain Hb. Addition of Ep to these small colonies led to the formation of erythroid bursts. Administration of Ep to the culture could be delayed for 6 days without decreasing the number of bursts if the cultures were initiated in the presence of BPA; in the absence of BPA, BFU-E were repidly lost if Ep was not provided at the start of the culture. BPA produced larger bursts than those formed in the presence of Ep alone. Microassays of Hb in the bursts indicated that BPA increased the amount of Hb per burst. This increase could not be entirely explained by the augumentation in cell number per burst but was partly ascribable to the increased amount of Hb. per cell.

红细胞生成需要两种不同的生长因子。促红细胞生成素（Ep）促进晚期红系前体细胞（CFU-E）的生长及其成熟为原成红细胞，其中血红蛋白（Hb）的合成开始。另一个因子，爆发促进活性（BPA）作用于早期前体细胞（BFU-E），并负责这些细胞分化为对Ep具有高反应性的CFU-E。本项目旨在从人源中纯化BPA，并探讨BPA和Ep这两种因子对甲基纤维素培养的人外周血单个核细胞红系爆发集落形成的影响。从再生障碍性贫血患者尿中分离出Ep，在去除Ep后的残留物中发现BPA。BPA的广泛纯化产生完全不含Ep的BPA制剂。仅添加Ep即可形成红色爆发。BPA的加入不仅增加了爆发的数量，而且大大减少了爆发形成所需的Ep量。在培养物中单独存在BPA不允许形成爆发，但允许不含Hb的小菌落生长。向这些小集落中加入Ep导致红系爆发的形成。如果在BPA存在的情况下开始培养，Ep的给药可以延迟6天而不减少爆发的数量;在没有BPA的情况下，如果在培养开始时不提供Ep，BFU-E会迅速丢失。BPA产生更大的爆发比那些形成的EP单独存在。对爆发中的Hb的微量测定表明，BPA增加了每次爆发的Hb量。这种增加不能完全解释为细胞数量的增加，但部分归因于增加量的Hb。每个细胞。