GROWTH FACTORS IN NORMAL AND NEOPLASTIC HEMATOPOIESIS

研究计划的目标是描述 造血生长因子，影响增殖， 早期人类祖细胞，包括正常的和肿瘤性的。 四个领域 将被强调。 首先，我们会继续研究 调节间充质细胞释放生长因子。 这将包括对各种生长因子的北方分析 从细胞如淋巴细胞、成纤维细胞、内皮细胞 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- （白细胞介素-3）集落刺激因子（CSF）将被 就业。 各种免疫介质的作用， 将研究炎症反应，包括 白细胞介素1，脂多糖，肿瘤坏死因子， 光敏素和凝血酶。 相对转录率 的基因的CSF将被测量，以确定他们是否 在不同的介质中有不同的表达 炎症 第二，我们将探讨结构/功能的关系， 人GM-和G-CSF在蛋白质和分子水平。 为 在蛋白质水平的研究，我们将产生单克隆 人GM-和G-CSF的抗体， 用这些因子的cDNA转染的哺乳动物细胞。 在 在分子水平上，将进行位点特异性诱变， 以确定基因的哪些元件对 完整蛋白质的生物学功能。 第三，我们将研究纯化的重组GM-和 G-CSF，单独和一起，对从 正常志愿者和患者的外周血和骨髓 骨髓增生性疾病 为了评估 生长因子与祖细胞的相互作用， 细胞将在无血清条件下以低浓度生长， 条件 第四，我们将寻求克隆cDNA， 猫白血病病毒感染产生的生长因子 胚胎猫成纤维细胞。 战略将涉及减法 杂交以富集生长因子的mRNA，产生 在表达载体中的cDNA文库，以及鉴定 通过生物学测定获得有用的克隆。

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.