CELLULAR MODULATION OF HEMOPOIESIS

了解人类造血调节的相关因素 需要评估相对均匀的细胞之间的相互作用 人口和纯增长因素。 我们利用细胞分离技术 提供获得显著富集的细胞的方法的步骤 来自骨髓的细胞群，其中异质细胞共存。 我们有 建立了用于检测增殖的体外克隆形成试验， 多能分化（CFU-GEMM）、红系分化（BFU-E）和 粒细胞-单核细胞（CFU-GM）人干细胞。 特别有用的是， 免疫学技术的加入， 回收抗原确定的造血细胞和细胞调节 它们的增殖和分化。 我们将扩大使用 免疫“淘选”（抗体介导的平板结合技术）， 结合密度和粘附分离程序，提供 相对同质的细胞群，用于产生刺激性或 抑制生长物质并提供这些物质的靶细胞 因素 此外，使用纯化的生长因子将允许更多的细胞生长。 精确描述监管相互作用，特别是 发生在骨髓造血微环境中。 我们计划 特别是为了扩展我们最近的研究， 与人类造血调节有关的生物学变量：（1） 刺激性和抑制性造血生长因子的产生 确定的免疫亚群;（2）检测生产和 粒细胞生成分化诱导因子的鉴定 （称为GM-CSF-D，分化因子-DF或MGI-2），与 以前定义明确的增殖诱导因子（GM-CSF-P），通过使用 能够被诱导分化的靶细胞（人 白血病细胞株HL-60）的骨髓靶向富集的反应性 细胞对多种免疫系统修饰剂（例如，白细胞介素），其他 物质和纯化的生长因子;和（4）的影响， “成熟诱导”物质，视黄酸和维生素D， 造血生长因子的产生和对造血生长因子的反应 包括干细胞自我复制。 这些研究将与 正常骨髓细胞以获得生理基线值， 来自骨髓性白血病、白血病前期、其他患者的细胞 骨髓增生性和粒细胞生成性疾病， 这些疾病的病理生理机制。 （IS）

Understanding factors involved in the regulation of human hematopoiesia requires evaluation of the interaction between relatively homogeneous cell populations and pure growth factors. We are utilizing cell separation procedures to provide methods for obtaining markedly enriched cell populations from marrow wherein heterogeneous cells coexist. We have established in vitro clonogenic assays for detecting proliferation and differentiation of pluripotent (CFU-GEMM), erythroid (BFU-E), and granulocyte-monocyte (CFU-GM) human stem cells. Of particular utility has been the addition of immunologic techniques to separate selectively and recover antigenically defined hematopoietic cells and cells modulating their proliferation and differentiation. We will extend our use of immunologic "panning" (an antibody-mediated plate-binding technique) in combination with density and adherence separation procedures to provide relatively homogeneous cell populations for generating stimulatory or inhibitory growth substances and to provide target cells for these factors. In addition, the use of purified growth factors will permit more precise delineation of regulatory interactions, particularly those occurring within the marrow hematopoietic microenvironment. We plan specifically to extend our recent studies and investigate several critical biologic variables involved with human hematopoietic regulation: (1) production of stimulatory and inhibitory hematopoietic growth factors by defined immunologic subsets; (2) detection of production and characterization of a differentiation-inducing factor for granulopoiesis (termed GM-CSF-D, differentiation factor-DF, or MGI-2), separate from the previously well-defined proliferation-inducing factor (GM-CSF-P), by use of target cells capable of being induced into differentiating (the human leukemic cell line HL-60); (3) the responsiveness of enriched marrow target cells to a variety of immune system modifiers (e.g., interleukins), other substances, and purified growth factors; and (4) the effects of the "maturation-inducing" substances, retinoic acid and vitamin D, on altering production of, and responsiveness to, hematopoietic growth factors including stem cell self-replication. These studies will be performed with normal marrow cells to obtain physiologic baseline values and with marrow cells from patients with myeloid leukemia, preleukemia, other myeloproliferative and granulopoietic disorders to assess possible pathophysiological mechanisms in these diseases. (IS)