CELLULAR AND HUMORAL MODULATION OF HEMOPOIESIS

我们将评估参与体液和细胞免疫的因素， 调节正常个体和患有 造血系统疾病，包括急性骨髓性白血病（AML）， 骨髓增生异常综合征（MDS）。 主要重点将放在 评价（1）纯化的之间的体外相互作用 重组人多肽造血生长因子 和人类造血细胞，和（2）富集的作用 辅助骨髓和血细胞群在生成HGF中的作用 在白细胞介素2（IL 2）和其他免疫调节剂的影响下， 调制器。 我们正在使用重组HGF文库， 单克隆抗体，并已开发出各种细胞 分离技术、配体结合程序和无血清 细胞增殖、分化和克隆形成测定， 进行这些研究。 关于（1）激素的具体目标 在特定细胞水平上， 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） 造血细胞增殖和分化，或者直接 或通过改变受体结合作用以刺激集落 因子（GM-CSF）或促红细胞生成素。 我们将评估假设 多种生长因子，包括胰岛素和IGFs， 原位提供细胞能力和进展因子， 造血由于造血祖细胞（HPC）的存活率是 部分通过HGF对葡萄糖转运和ATP的作用介导 我们将研究这一代人之间的相互作用， 代谢活性促有丝分裂激素胰岛素、IGF和CSF， 探讨造血调控机制。 研究使用 补充的无血清培养基将用人骨髓细胞进行 红系细胞系和富集的HPC作为靶细胞， 精确确定功能效果和关系 在HGF之间 此外，我们将评估这些细胞产生IGFs的情况。 细胞，以评估可能的自分泌或旁分泌的作用， 这些激素对白血病细胞和正常细胞生长的影响。 (2)我们将 检查IL 2在通过富集HGF来增强HGF提供中的作用 T细胞和非T细胞亚群，特别强调效应细胞 自然杀伤（NK）细胞。 我们将评估产量 增殖因子（GM-CSF，BPA）和分化因子（G-CSF， DF）与静息T细胞和NK细胞（缺乏TAC+ IL 2受体）相比 这种活化的Tac+）细胞以评估IL 2受体的作用 产生HGF。 在标准化技术以定义HGF之后， 正常骨髓细胞的产生、结合和反应性， 白血病细胞系，我们将（3）评估这些参数， 来自造血系统疾病患者的细胞，特别是AML 和MDS，并将根据这些基本的 细胞生长调节缺陷。

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.