LEUKEMIA INHIBITOR SUPPRESSES DIFFERENTIATION & IMMUNITY

白血病抑制剂抑制分化

• 批准号: 3173550
• 负责人: J. W. none CHIAO
• 金额: $ 18.21万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-03-01 至 1990-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3173550
• 关键词: B lymphocyte; cell differentiation; cell growth regulation; cellular oncology; cytotoxicity; human tissue; leukocyte antigen typing; leukopoietic factor; myelogenous leukemia; neoplasm /cancer diagnosis; tissue /cell culture

We have successfully induced leukemic cells from myelogenous leukemia patienta and HL-60 promyelocyte-cell line to differentiate and mature to macrophages in a liquid culture assay. Conditioned culture medium (CM) of PHA-activated normal T cells contains the maturation-inducing activity. Analysis of total DNA content of single cells by flow cytometry demonstrated clearly that differentiated macrophages possess the same aneuploid DNA stemline as the undifferentiated patient cells, indicating leukemic cells can differentiate independently of genetic abnormality. The developmental process revealed that CM first induced most of the leukemic cells for one cell cycle division, then suppressed the DNA replication preceding mature cell expression. This process has also been delineated by reduction of RNA content, development of adherent cells, morphology, and acquisition of membrane receptors and functions. We hope these analyses of proliferation and maturation will represent a new and practical method to study leukemia. We are intending to study leukemic-patient cells with simultaneous application of the methods probing proliferation and differentiation defects. This includes the investigation of a lack of capacity to stop leukemic cells from replicating by comparing the suppressive activity of CM of patients with normal proliferation of HL-60 cells. Microassay technique to quantitate replicating cell number by autoradiography will be used. A possible excess or lack of proliferative stimulation will be studied by cross-testing CM of patients and of normal controls with patient and normal cells. Similar methods will be used with the culture assay to analyze whether there is a lack of CM maturation-inducing activity for their own cells, thereby blocking their differentiation. The development of isolated immature cells of normal bone marrow could be compared. We also intend to identify the subset of T cells producing the activities for cell proliferation, differentiation, and suppression of proliferation. Combining assays of proliferation and differentiation on the same cells will enable us to understand the controls of the leukemic cell growth. (MI)

我们成功地从骨髓性白血病中诱导出白血病细胞 Patienta 和 HL-60 早幼粒细胞系分化并成熟为 液体培养测定中的巨噬细胞。 条件培养基 (CM) PHA 激活的正常 T 细胞具有成熟诱导活性。 流式细胞术分析单细胞总DNA含量 清楚地表明分化的巨噬细胞具有相同的 非整倍体 DNA 干系作为未分化的患者细胞，表明 白血病细胞可以独立于遗传异常而分化。 这 发育过程显示，CM首先诱发了大部分白血病 细胞进行一个细胞周期分裂，然后抑制DNA复制 成熟细胞表达之前。 这个过程也被描述为 RNA含量减少、贴壁细胞发育、形态学和 获得膜受体和功能。 我们希望这些分析 扩散和成熟将代表一种新的、实用的方法 研究白血病。 我们打算研究白血病患者细胞 同时应用探测增殖和 分化缺陷。 这包括调查缺乏 通过比较阻止白血病细胞复制的能力 HL-60 增殖正常的患者的 CM 抑制活性 细胞。 微量测定技术通过以下方法定量复制细胞数量 将使用放射自显影术。 可能存在过度或缺乏增殖 将通过对患者和正常人的 CM 进行交叉测试来研究刺激 用患者和正常细胞进行对照。 类似的方法将用于 培养检测分析是否存在CM缺乏 自身细胞的成熟诱导活性，从而阻止它们 差异化。 正常骨分离未成熟细胞的发育 骨髓可以比较。 我们还打算鉴定 T 细胞的子集 产生细胞增殖、分化等活性 抑制增殖。 结合增殖分析和 相同细胞的分化将使我们能够理解控制 白血病细胞的生长。 （MI）