GROWTH FACTORS IN NORMAL AND NEOPLASTIC HEMATOPOIESIS

正常和肿瘤造血中的生长因子

• 批准号: 3169705
• 负责人: JOHN W ADAMSON
• 金额: $ 15.24万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3169705
• 关键词: RNA; binding proteins; cell growth regulation; cellular oncology; colony stimulating factor; complementary DNA; embryo /fetus tissue /cell culture; enzyme linked immunosorbent assay; feline leukemia /sarcoma virus; fibroblasts; gel electrophoresis; gene expression; genetic manipulation; genetic transcription; glucose 6 phosphate dehydrogenase deficiency; glycosylation; granulocyte; hematopoiesis; hematopoietic stem cells; high performance liquid chromatography; human subject; human tissue; hybridomas; immunomodulators; interferons; interleukin 1; interleukin 3; laboratory mouse; laboratory rabbit; lipopolysaccharides; macrophage; molecular cloning; molecular oncology; monoclonal antibody; myeloproliferative neoplasm; neoplastic growth; nucleic acid probes; oligonucleotides; point mutation; protein biosynthesis; thrombin; transfection; tumor necrosis factor alpha; tumor necrosis factor beta; ultracentrifugation; vascular endothelium

The goals of the research program are to characterize hematopoietic growth factors which influence the proliferation of early human progenitors, both normal and neoplastic. Four areas will be emphasized. First, we will continue studies of the regulation of growth factors release form mesenchymal cells. This will include Northern analyses of the kinds of growth factors released from cells such as lymphocytes, fibroblasts, endothelial cells, and smooth muscle cells. Probes for human granulocyte /macrophage (GM), granulocyte (G), macrophage (M) and multi- (interleukin-3) colony-stimulating factors (CSFs) will be employed. The effects of various mediators of immune and inflammatory responses will be studied and will include interleukin1, lipoppolysaccharide, tumor necrosis factor, lymphotoxin, and thrombin. Relative transcription rates of the genes for the CSFs will be measured to determine if they are differentially expressed by the various mediators of inflammation. Second, we will explore the structure/function relationship of human GM- and G-CSF at the protein and molecular levels. For studies at the protein level, we will generate monoclonal antibodies to human GM- and G-CSF by purifying them from mammalian cells transfected with cDNA for these factors. At the molecular level, sitespecific mutagenesis will be carried out to determine which element of the gene are important for biological function of the intact protein. Third, we will study the effect of purified recombinant GM- and G-CSF, alone and together, on progenitor cells obtained from the peripheral blood and marrow of normal volunteers and patients with myeloproliferative disorders. To assess the nature of the interaction of the growth factors with the progenitor cells, the cells will be grown at low concentrations under serumfree conditions. Fourth, we will seek to clone the cDNA for a potentially novel growth factor produced by feline leukemia virusinfected embtyonic cat fibroblasts. The strategy will involve subtraction hybridization to enrich mRNA for the growth factor, generation of a cDNA library in an expression vector, and identification of useful clones by biological assays.

研究计划的目标是确定 影响骨髓间充质干细胞增殖的造血生长因子 早期的人类祖先，包括正常的和肿瘤的。四个领域 将会被强调。首先，我们将继续研究 间充质细胞释放生长因子的调控。 这将包括对各种增长因素的诺斯分析 从淋巴细胞、成纤维细胞、内皮细胞等细胞释放 细胞和平滑肌细胞。人粒细胞探针 巨噬细胞(GM)、粒细胞(G)、巨噬细胞(M)和多核细胞 (IL-3)集落刺激因子(CSF) 受雇的。不同免疫调节因子对机体免疫功能的影响 将研究炎症反应，并将包括 白介素1、脂多糖、肿瘤坏死因子、 淋巴毒素和凝血酶。相对转录速率 将对CSF的基因进行测量以确定它们是否 由不同的调解人表达的不同 发炎。 第二，我们将探索结构/功能关系 人GM-和G-CSF在蛋白质和分子水平的表达。为 在蛋白质水平上的研究，我们将产生单克隆 纯化抗人GM-和G-CSF抗体的研究 将这些因子的c DNA导入哺乳动物细胞。在… 将在分子水平上进行定点突变 来确定基因中的哪些元素对 完整蛋白质的生物学功能。 第三，我们将研究纯化的重组GM-And的作用 G-CSF单独和共同作用于从人外周血中获得的祖细胞 正常志愿者和患者的外周血和骨髓 患有骨髓增生性疾病。以评估该事件的性质 生长因子与祖细胞的相互作用 细胞将在低浓度的无血清条件下生长 条件。 第四，我们将试图克隆一个潜在的小说的cdna 猫白血病病毒感染产生的生长因子 胚胎猫成纤维细胞。这一策略将涉及减法 杂交富集生长因子的mRNA代 在表达载体中构建一个cDNA库，并对其进行鉴定 通过生物检测获得有用的克隆。