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

描述：（改编自研究者摘要）。 粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白介素-3 IL-3是造血生长因子（HGF）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协同作用。 我们的长期目标是 为了了解介导GM-CSF和IL-3的分子事件， 相互交叉竞争结合到细胞表面， 这两种受体;这发生在4摄氏度，表明它可能是 由于细胞表面的相互作用。 交联研究表明， GM-CSF和IL-3受体都是复杂的异二聚体或 异源三聚体蛋白。 我们希望验证高亲和力 GM-CSF和IL-3的结合需要每种受体的表达， 一个独特的和共同的亚基;以及表达这两种受体的细胞上的亚基 它们相互作用形成由共同亚基连接的GM-CSF/IL-3复合物。 我们计划通过分离编码 GM-CSF和IL-3受体亚单位。 我们将研究结构 通过转染突变体的受体亚基的功能关系 因子依赖性鼠细胞系中，并通过抗肽单克隆抗体 抗体的 我们的目标是确定高/低的结构要求 通过将cDNA共转染入 受体阴性细胞。 我们计划鉴定出与 磷蛋白与受体结合后的受体 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， Kostmann和Diamond Blackfan贫血 我们将使用PCR方法 家系外连锁分析及序列推定突变体 受体。 我们认为，这些研究HGF受体的功能， 正常和不正常的个体将提供深入了解分子 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.