CHARACTERIZATION OF ACUTE MYELOGENOUS LEUKEMIA STEM CELL

该项目涉及白血病集落形成细胞的研究。 急性髓系白血病患者的群体(L-氟氯化碳)。在……里面 本研究的一个方面，L-CFCs表达的细胞表面抗原 将在急性髓系白血病患者中使用一组髓系特异性的 单抗和补体介导的细胞毒作用。患者会 在随后的复发时进行连续检查，以确定L-氟氯化碳 表型随时间变化稳定。这些研究的结果将在那时 与一系列旨在检验诱导性的实验相比较 用多种已知作用于髓系的药物促进L-CFCs的成熟 细胞分化。接受检测的试剂包括重组伽马 干扰素、1，25二羟基维生素D3和顺式维甲酸。中的更改 将检查表面抗原、增殖能力和形态。 L-氯氟化碳暴露在最佳浓度下的成熟诱导作用 探员们。这些研究旨在检查急性髓细胞白血病中的L-氟氯化碳是否 对分化因子作出反应，并确定是否有反应 表现为与正常骨髓生成相似的细胞变化 (即，更分化的细胞表型的表达和减少 在细胞增殖中)。 另一个方面是将两者的细胞表面抗原性展示相关联 有癌基因表达的正常和白血病髓系细胞。表达方式 将研究三种髓系相关细胞癌基因(c-onc)： N-ras、c-myc和c-fos。癌基因表达将通过以下方式进行量化 从正常髓系提取的c-onc RNA的探针杂交 亚群(单核细胞和中性粒细胞)和白血病原始细胞 来自急性髓系白血病患者的人群。癌基因蛋白的表达 产品将通过使用抗血清进行免疫荧光研究来确定 癌基因蛋白，并通过免疫沉淀研究放射性标记的 细胞裂解物。这些研究的目的是关联基因的表达 正常和非癌组织中髓系细胞表面抗原模式的癌基因 判断恶性细胞是否与 差异化状态。结果将加深我们对白血病的理解 AML中的细胞生物学，并旨在阐明成熟的 髓系白血病中出现的缺陷。通过确定 L-氟氯化碳人口的成熟能力、替代办法 治疗，包括免疫治疗和成熟诱导治疗，可以 结果。

This project involves the study of the leukemia-colony forming cell population (L-CFC) in patients with acute myelogenous leukemia (AML). In one aspect of this study, the cell surface antigens expressed by the L-CFC will be examined in patients with AML using a panel of myeloid-specific monoclonal antibodies and complement-mediated cytotoxicity. Patients will be examined serially on subsequent relapses to determine whether the L-CFC phenotype is stable with time. The findings from these studies will then be compared to a series of experiments designed to examine the inducibility to maturation of the L-CFC using various agents known to effect myeloid cell differentiation. The agents to be tested include recombinant gamma interferon, 1,25 dihydroxyvitamin D3, and cis-retinoic acid. Changes in surface antigens, proliferative potential, and morphology will be examined in the L-CFC exposed to optimal concentrations of the maturation-inducing agents. The studies are designed to examine whether the L-CFC in AML is responsive to agents of differentiation, and to determine whether response is manifested in cellular changes similar to those of normal myelopoiesis (i.e., expression of a more differentiated cell phenotype and a reduction in cellular proliferation). Another aspect is to correlate the cell surface antigenic display of both normal and leukemic myeloid cells with oncogene expression. The expression of three myeloid-associated cellular oncogenes (c-onc) will be studied: N-ras, c-myc and c-fos. Oncogene expression will be quantified by hybridization of probes to c-onc RNA extracted from normal myeloid subpopulations (monocytes and neutrophils) and from leukemia blast cell populations from patients with AML. Expression of oncogene protein products will be determined by immunofluorescence studies using antisera to the oncogene proteins, and by immunoprecipitation studies of radiolabelled cell lysates. The aim of these studies is to correlate the expression of oncogenes with patterns of myeloid cell-surface antigens on normal and malignant cells to determine whether there is a relationship with differentiation status. Results will further our understanding of leukemia cell biology in AML, and are directed towards clarifying the maturational defect that occurs in myelogenous leukemia. By determining the maturational capabilities of the L-CFC population, alternative approaches to therapy, including immunotherapy and maturation-inducing therapy, may result.