IN VITRO STUDIES OF HEMATOPOIETIC REGULATION

这项研究涉及到一些方面的造血调节。 特别强调荷尔蒙和细胞的体外评估 互动。具体研究概括在三个方面： 1.造血细胞生长中的激素和细胞相互作用。我们 将研究血小板衍生生长因子(PDGF)和 红系骨髓基质细胞释放的生长因子， 人粒细胞/巨噬细胞和巨核细胞集落形成。这个 这些相互作用的靶细胞(早期或晚期红系祖细胞)和 将定义潜在的附属单元。体外抑制物 慢性肾功能衰竭(CRF)的造血功能将被评估并 与尿毒症和贫血的程度相关 肾功能的范围。我们还将治疗一些慢性肾功能衰竭患者 富含促红细胞生成素(EP)的人血浆定量评估其 回应。被发现具有活性的抑制剂将被部分分离出来，并 他们对造血细胞生长的影响进行了更详细的研究 我们使用了人类和小鼠的造血细胞系。 2.EP提纯。人类尿液EP量的增加将是 纯化的抗体被提升到氨基末端序列以及 纯化的激素，并进行免疫亲和层析以增加 活性荷尔蒙的恢复。从贫血的狒狒肾脏中提取的mRNA已经被 提取并制备了cDNA文库。我们将采取各种策略 为了标记纯化的激素，准备放射免疫分析，并鉴定 含有EP编码序列的基因组材料的克隆。 3.造血多能干细胞克隆性障碍。我们会 继续使用葡萄糖-6-磷酸脱氢酶(G6PD)作为标志物 关注原发性血小板增多症患者，并使用人类 巨核细胞集落形成细胞试验以及一种 多能祖细胞。

The studies address aspects of the regulation of hematopoiesis with specific emphasis on the in vitro assessment of hormonal and cellular interactions. Specific studies are outlined in three general areas: 1. Hormonal and cellular interactions in hematopoietic cell growth. We will study the influence of platelet derived growth factor (PDGF) and growth factors released by marrow stromal cells on erythroid, granulocyte/macrophage, and megakaryocytic colony formation in man. The target cell for these interactions (early or late erythroid progenitor) and potential accessory cells will be defined. Inhibitors of in vitro hematopoieses in chronic renal failure (CRF) will be assessed and correlated with the extent of uremia and anemia in patients with a wide range in renal function. We will also treat some patients with CRF with erythropoietin (Ep)-rich human plasma to quantitatively assess their response. Inhibitors found to be active will be partially isolated and a more detailed study of their effect on hematopoietic cell growth carried out employing human and murine hematopoietic cell lines. 2. Ep purification. Increased amounts of human urinary Ep will be purified, antibodies raised to the amino terminal sequences as well as to purified hormone, and immunoaffinity chromatography performed to increase recoveries of active hormone. mRNA from anemic baboon kidney has been extracted and a cDNA library prepared. Various strategies will be employed to label the purified hormone, prepare a radioimmunoassay, and identify clones of genomic material containing Ep-coding sequences. 3. Clonal disorders of the hematopoietic multipotent stem cell. We will continue to use glucose-6-phosphate dehydrogenase (G6PD) as a marker focusing on patients with essential thrombocytosis and employing the human megakaryocyte colony-forming cell assay as well as an assay for a multipotent progenitor.