CELLULAR MODULATION OF HEMOPOIESIS

我们将评估与体液和细胞有关的因素 正常人和慢性阻塞性肺疾病患者的造血调控 造血疾病，包括急性髓系白血病(AML)和 骨髓增生异常综合征(MDS)。主要重点将放在 评价(1)纯化的人血清白蛋白在体外的相互作用 重组人多肽造血生长因子(HGFs) 和人的造血细胞，以及(2)浓缩的作用 人成纤维细胞生长因子生成中的副骨髓和血细胞群体 在白介素2(IL2)和其他免疫的影响下 调制器。我们正在使用一个重组HGFs文库， 并已研制出多种单抗和细胞 分离技术、配体结合程序和无血清 细胞增殖、分化和克隆形成试验 进行这些研究。关于(1)荷尔蒙的具体目标 相互作用，是在特定细胞水平上确定的 表面受体，中胚层刺激激素的作用 胰岛素和胰岛素样生长因子(IGFI&II)的调节 造血细胞的增殖和分化，或者直接 或通过改变受体的结合作用来刺激集落 因子(GM-CSF)或促红细胞生成素。我们将评估这一假说 包括胰岛素和胰岛素样生长因子在内的多种生长因子相互作用 在现场提供细胞能力和进展因子 造血术。随着造血祖细胞(HPC)的存活 肝细胞生长因子对葡萄糖转运和三磷酸腺苷的影响 一代，我们将研究 代谢活性促有丝分裂激素胰岛素、IGFS和CSF 探讨造血调控的机制。研究使用 补充的无血清培养液将与人髓系一起进行 并以红系细胞和富集型HPC为靶细胞，以更多 准确确定功能效果和关系 在HGFs之间。此外，我们将通过以下方式评估IGFS的产量 细胞以评估其可能的自分泌或旁分泌作用 这些荷尔蒙对白血病和正常细胞生长有促进作用。(2)我们会 研究IL2在通过富集化促进HGFs供应方面的作用 T和非T细胞亚群，特别强调效应器 自然杀伤(NK)细胞功能。我们将对产量进行评估 增殖因子(GM-CSF、BPA)和分化因子(G-CSF、 通过静息T和NK细胞(缺乏TAC+IL2受体)与 这种被激活的Tac+细胞评估IL2受体的作用 在生成HGFs方面。在标准化技术定义HGF之后 正常髓系细胞产生、结合和反应 白血病细胞株，我们将(3)评估这些参数 来自我们的造血疾病患者的细胞，特别是急性髓系白血病 和MDS，并将根据这些基本情况对这些疾病进行分类 生长调节中的细胞缺陷。

We will evaluate factors involved in the humoral and cellular modulation of hemopoiesis in normal individuals and those with hemopoietic diseases, including acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). Major emphasis will be placed on evaluating (1) the in vitro interactions between purified recombinant human polypeptide hemopoietic growth: factors (HGFs) and human hemopoietic cells, and (2) the roles of enriched populations of accessory marrow and blood cells in generating HGFs under the influence of interleukin 2 (IL2) and other immune modulators. We are using a library of recombinant HGFs and monoclonal antibodies and have developed a variety of cell separation techniques, ligand binding procedures and serum-free cell proliferation, differentiation and clonogenic assays to perform these studies. Specific objectives regarding (1) hormonal interactions, are to determine, at the level of specific cell surface receptors, the role of the mesodermal stimulatory hormones insulin and insulin-like growth factors (IGFI & II) for modulating hemopoietic cell proliferation and differentiation, either directly or via alteration of receptor binding action for colony stimulating factor (GM-CSF) or erythropoietin. We will evaluate the hypothesis that multiple growth factors, including insulin and IGFs, interact in situ to provide cellular competence and progression factors for hemopoiesis. As hemopoietic progenitor cell (HPC) survival is partly mediated through HGF effects on glucose transport and ATP generation, we will examine the interactions between the metabolically active mitogenic hormones insulin, IGFs and CSFs to probe mechanisms of hemopoietic regulation. Studies using supplemented serum-free medium will be performed with human myeloid and erythroid cell lines and enriched HPCs as target cells to more precisely determine the functional effects and relationships between HGFs. Further, we will assess production of IGFs by these cells to evaluate the possible autocrine or paracrine roles of these hormones for leukemic vs normal cell growth. (2) We will examine the role of IL2 in enhancing provision of HGFs by enriched T and non T- cell subsets, with particular emphasis on the effector function of natural killer (NK) cells. We will assess production of proliferative (GM-CSF, BPA) and differentiative factors (G-CSF, DF) by resting T and NK cells (which lack TAC+ IL2 receptors) vs such activated Tac+) cells to evaluate the role of IL2 receptors in generating HGFs. After standardizing techniques to define HGF production, binding and responsiveness by normal myeloid cells and leukemic cell lines, we will (3) evaluate these parameters for cells from our patients with hemopoietic diseases, particularly AML and MDS, and will classify these disorders according to such basic cellular defects in growth regulation.