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。 重组 蛋白质的生产和表征。 虽然很多 在基因和蛋白质水平上了解这些因素， 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 他们的行动机制。 我们计划调查 重要的是，通过比较已知产生HGF的细胞， 在未受刺激的情况下综合不同因素的能力 条件和用各种诱导剂刺激后。 这将通过北方印迹分析RNA来完成， 蛋白质免疫印迹和免疫沉淀， GMCSF的情况下，通过放射免疫测定，以及。 我们打算 鉴定组织切片和细胞离心涂片制备物中的生产细胞 利用强大的原位RNA 杂交技术结合免疫酶 染色 我们将开始研究 通过使用高度纯化的放射性标记的GMCSF来研究HGF 受体结合正常和白血病细胞。我们的长期 目的是通过电泳来表征受体， 含有交联受体的增溶膜， 放射性标记的GMCSF。 最后，我们希望克隆基因， 通过使用生物素化的GMCSF，荧光 荧光激活细胞分选技术 用基因组转化后表达受体的细胞 DNA. 我们坚信，这些研究将构成 为制定合理的标准采取重要步骤， GMCSF和其他生长因子的治疗用途， 变得可用。 此外，我们希望了解 增殖和分化的机制是 在正常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.