SOURCES AND ACTIONS OF HUMAN GMCSF AND MULTI-CSF

在过去的两年里，五个人类的基因 造血生长因子已被克隆出来。重组 制备了蛋白质并对其进行了鉴定。虽然很多都是 在基因和蛋白质水平上已知这些因素，有 关于合成它们的细胞的识别信息较少， 调节合成的生理机制，以及 它们的作用机制。我们计划调查他的亲属 通过比较已知的产生肝细胞生长因子的细胞 在非刺激条件下合成不同因素的能力 条件和刺激后用各种诱导剂。 这将通过Northern blots分析RNA来实现， 免疫印迹和免疫沉淀中的蛋白质，以及 GMCSF 1例，用放射免疫法测定。我们打算 鉴定组织切片和细胞旋体制剂中的产生细胞 利用强大的原位RNA进行造血器官的研究 与免疫酶结合的杂交技术 染色。我们将开始研究其作用机制。 高纯度放射性标记巨噬细胞集落刺激因子对肝细胞生长因子的影响 正常和白血病细胞上的受体结合。我们的长期目标 目的是通过电泳法鉴定受体的特性。 含有交联型受体的增溶膜 放射性标记的GMCSF。最后，我们希望克隆该基因 利用生物素标记的GMCSF受体，荧光 微球与荧光激活的小鼠L细胞分选 用基因组转化后表达受体的细胞 DNA我们相信，这些研究将构成 定义合理的标准的重要步骤 巨噬细胞集落刺激因子和其他生长因子的治疗应用 变得有空。此外，我们希望深入了解 增殖和分化的机制 在正常细胞和白血病细胞中控制。

During the past two years, the genes for five human hematopoietic growth factors have been cloned. Recombinant proteins were produced and characterized. Although much is known about these factors at a gene and protein level, there is less information on the identify of the cells that synthesize them, the physiological mechanism by which synthesis is regulated, and their mechanism of action. We plan to investigate the relative important of cells known to produce HGF's by comparing their capacity to synthesize the different factors under unstimulated conditions and after stimulation with a variety of inducing agents. This will be accomplished by analyzing RNA by Northern blots, protein in Western blots and immunoprecipitations and, in the case of GMCSF, by radioimmune assay, as well. We intend to identify producer cells in tissue sections and cytospin preparations of hematopoietic organs by using powerful in situ RNA hybridization techniques combined with immunoenzymatic staining. We will begin to investigate the mechanism of action of HGF's by using highly purified radiolabeled GMCSF to study receptor binding on normal and leukemic cells. Our longer term aims are to characterize the receptor by electrophoresis of solubilized membranes containing receptor crosslinked to radiolabeled GMCSF. Finally, we wish to clone the gene for the GMCSF receptor by using biotinylated GMCSF, fluorescent microsphere and fluorescence activated cell sorting of murine L cells that express the receptor after transformation with genomic DNA. It is our conviction that these studies will constitute important steps toward defining rational criteria for the therapeutic use of GMCSF and the other growth factors as they become available. Furthermore, we hope to gain insight into the mechanism by which proliferation and differentiation are controlled in normal and leukemic cells.