THE SOURCES AND ACTIONS OF HUMAN GM-CSF AND MULTI-CSF

描述：(改编自调查人员摘要)。 粒细胞巨噬细胞集落刺激因子与白细胞介素3 白介素3(IL-3)是造血生长因子家族的两个成员。 它们刺激原始细胞的增殖和分化 造血干细胞和成熟细胞的功能；此外， 它们在体外和体内都有协同作用。我们的长期目标是 了解介导GM-CSF和IL-3表达的分子事件 与细胞表面结合的互惠交叉竞争 两种受体；这发生在零下4摄氏度，表明它可能是 由于细胞表面的相互作用。交联性研究表明， GM-CSF和IL-3受体都是复杂的异二聚体或 异三聚体蛋白质。我们希望检验高亲和力的假设 GM-CSF和IL-3的结合需要对每个受体表达一种 唯一和共同的亚基；以及在表达这两种受体的细胞上的 它们相互作用，形成由共同亚基连接的GM-CSF/IL-3复合体。 我们专门计划通过分离编码的cDNA来检验这一假说 GM-CSF是IL-3受体的一个亚单位.我们将调查结构 转导突变体后受体亚基的功能关系 转化成因子依赖的小鼠细胞系，并用抗肽单抗 抗体。我们的目标是确定高/低的结构要求 共转染cDNAs的亲和结合和交叉竞争 受体阴性细胞。我们计划找出与之相关的蛋白质 磷蛋白与受体配基结合后的受体 免疫沉淀分析。最后，我们的目标是研究受体 遗传性骨髓衰竭综合征患者的功能 科斯特曼和戴蒙德·布莱克凡贫血。我们将使用聚合酶链式反应的方法进行 家系外源连锁分析及序列推测突变 感受器。我们认为，这种对HGF受体功能的研究在这两种情况下 正常和不正常的个体将提供对分子的洞察 调节造血，并导致新的治疗可能性。

DESCRIPTION: (Adapted from investigator's abstract). Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-3 (IL-3) are two members of a family of hematopoietic growth factors (HGFs). They stimulate the proliferation and differentiation of primitive hematopoietic stem cells, and the function of mature cells; furthermore, they act synergistically both in vitro and in vivo. Our long-term goal is to understand the molecular events that mediate GM-CSF and IL-3 show reciprocal cross competition for binding to the surface of cells that bear both receptors; this occurs at 4 degrees-C, indicating that it is probably due to interaction at the cell surface. Crosslinking studies suggest that both the GM-CSF and the IL-3 receptors are complex heterodimeric or heterotrimeric proteins. We wish to test the hypothesis that high affinity binding of GM-CSF and IL-3 requires expression, for each receptor, of a unique and a common subunit; and that on cells that express both receptors they interact to form a GM-CSF/IL-3 complex linked by the common subunit. We plan specifically to test this hypothesis by isolating cDNAs that encode the GM-CSF an IL-3 receptor subunits. We will investigate structure function relationships of the receptor subunits by transfection of mutants into factor-dependent murine cell lines and by antipeptide monoclonal antibodies. We aim to determine the structural requirements for high/low affinity binding and cross competition by cotransfection of cDNA's into receptor negative cells. We plan to identify proteins that associate with the receptor after ligand binding by phosphoprotein and receptor immunoprecipitation analyses. Last, we aim to investigate receptor function in patients with inherited bone marrow failure syndromes such as Kostmann's and Diamond Blackfan anemia. We will use PCR methods to carry out linkage analysis in affected families and sequence putative mutant receptors. We believe that such studies of HGF receptor function in both normal and abnormal individuals will provide insight into the molecular regulation of hematopoiesis and lead to new therapeutic possibilities.