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

人类 GM-CSF 和多种 CSF 的来源和作用

• 批准号: 3188673
• 负责人: COLIN A SIEFF
• 金额: $ 22.16万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-01 至 1996-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3188673
• 关键词: T lymphocyte; antibody receptor; antiserum; autoradiography; binding proteins; bone marrow; bone marrow disorder; bone marrow transplantation; cell bank /registry; cell differentiation; cell growth regulation; cell type; colony stimulating factor; complementary DNA; embryo /fetus cell /tissue; flow cytometry; gel electrophoresis; gene expression; genetic manipulation; granulocyte; growth factor receptors; hematopoietic growth factor; hematopoietic stem cells; human subject; human tissue; immunochemistry; interleukin 3; laboratory rabbit; ligands; molecular cloning; monoclonal antibody; mutagens; neoplastic transformation; northern blottings; nucleic acid probes; nucleic acid sequence; phosphoproteins; polymerase chain reaction; protein biosynthesis; radioimmunoassay; receptor; transfection

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.

描述：（根据研究者的摘要进行了改编）。 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具有协同作用。 我们的长期目标是 了解介导GM-CSF和IL-3显示的分子事件 互惠交叉竞争，以结合轴承的细胞表面 两个受体；这发生在4度C，表明它可能是 由于在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 异三聚合物蛋白。 我们希望检验高亲和力的假设 GM-CSF和IL-3的结合需要表达每个受体的表达 独特和一个共同的亚基；并且在表达两个受体的细胞上 它们相互作用以形成由公共亚基链接的GM-CSF/IL-3复合物。 我们计划通过隔离编码的cDNA来专门检验该假设 GM-CSF和IL-3受体亚基。 我们将研究结构 通过转染突变体的受体亚基的功能关系 进入因子依赖性鼠细胞系和抗肽单克隆 抗体。 我们旨在确定高/低的结构要求 通过共转染cDNA的亲和力结合和交叉竞争 受体负细胞。 我们计划确定与 配体结合后通过磷蛋白和受体结合的受体 免疫沉淀分析。 最后，我们旨在调查受体 遗传性骨髓衰竭综合症患者的功能，例如 科斯曼和钻石黑凡贫血。 我们将使用PCR方法携带 在受影响的家庭和推定突变体的序列分析中分析 受体。 我们认为，这两种HGF受体功能的这种研究 正常和异常个体将洞悉分子 调节造血并导致新的治疗可能性。