DIFFERENTIATIVE PROGRAMS OF LYMPHOID PROGENITOR CELLS

淋巴祖细胞的分化程序

• 批准号: 3169773
• 负责人: Tucker W LeBien
• 金额: $ 20.66万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 1995-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3169773
• 关键词: Abelson leukemia virus; B lymphocyte; CD antigens; DNA directed DNA polymerase; SDS polyacrylamide gel electrophoresis; acute lymphocytic leukemia; autoradiography; cell differentiation; cell growth regulation; clone cells; cytokine; disease /disorder model; flow cytometry; fluorescence microscopy; gel electrophoresis; gene expression; gene rearrangement; hematopoietic stem cells; host neoplasm interaction; human tissue; immunofluorescence technique; immunoglobulin genes; immunoregulation; leukocyte activation /transformation; leukopoiesis; lymphocytic leukemia; membrane proteins; molecular cloning; molecular oncology; monoclonal antibody; neoplastic transformation; neuropeptides; neutrophil; nucleic acid sequence; oncogenes; pediatric neoplasm /cancer; peptidases; polymerase chain reaction; protein sequence; receptor binding; tissue /cell culture; transforming virus

The major objective of this project is to study the cellular and molecular events associated with the differentiation of normal and malignant human lymphoid progenitor cells (LPC). The study will emphasize the clonal expansion of progenitor cells in non-T, non-B acute lymphoblastic leukemia (ALL), and their nonmalignant counterparts present in bone marrow (BM). Emphasis will be placed on the use of three monoclonal antibodies recently produced in our laboratory, designated BA-1, BA-2, and BA-3, that appear to bind to normal and malignant LPC at various stages of lymphoid cell development. Four specific aims have been or are being pursued: (1) A large number of lymphohematopoietic cell sources have been analyzed to determine the relationship between the aforementioned monoclonal antibodies and two intracellular markers of LPC, i.e., terminal deoxynucleotidyl transferase (TdT) and cytoplasmic IgM (CIgM). The cell populations were assayed by single or double fluorochrome staining using fluorescent microscopy and flow cytofluorimetry. The results of these studies, which identified candidate lymphoid progenitor cells, have been published. (2) The phorbol ester TPA has been used to study the induction of differentiation in normal and malignant LPC. TPA-induced changes were analyzed by immunofluorescence and biosynthetic labeling followed by SDS-PAGE. We have attempted to determine if there is a sequential (differentiation-linked?) expression of cell surface structures recognized by monoclonal antibodies and their relationship to the intracellular markers of LPC, i.e., TdT and CIgM. The results suggest that acquisition/loss of certain markers may be consistent with differentiation-linked events in lymphoid cell precursors. (3) Major emphasis is currently placed on examining the function that cell surface molecules gp40/BA-I, p24/BA-2, and gp100/CALLA/BA-3 subserve to the cells on which they are expressed. (4) Of particular interest is our recent observation that gp100/CALLA is strikingly decreased on the surface of neutrophils from patients with severe thermal injuries. Since neutrophils from these patients exhibit various dysfunctions (i.e., in hemotaxis), we are examining the possible role of gp100/CALLA in neutrophil function. (4) Studies are underway to clone the genes encoding gp40/BA-1, p24/BA-2, and gp/100/CACCA/BA-3 using mouse L cell transfection and subtractive hybridization. (LB)

该项目的主要目的是研究细胞和分子 与正常和恶性人的分化相关的事件 淋巴样祖细胞（LPC）。 该研究将强调克隆 非-T非-B急性淋巴细胞白血病中祖细胞的扩展 （全部）及其在骨髓（BM）中存在的非恶性对应物。 最近将重点放在三种单克隆抗体上 在我们的实验室中生产，指定为BA-1，BA-2和BA-3，似乎是 在淋巴样细胞的各个阶段与正常和恶性LPC结合 发展。 已经或正在追求四个具体目标：（1） 已经分析了大量的淋巴瘤性细胞来源 确定上述单克隆抗体之间的关系 LPC的两个细胞内标记，即末端脱氧核苷酸 转移酶（TDT）和细胞质IgM（CIGM）。 细胞群是 用荧光染色单或双荧光色素染色测定 显微镜和流式细胞荧光法。 这些研究的结果， 已确定的候选淋巴样祖细胞已发表。 （2） 凤凰酯TPA已用于研究诱导 正常和恶性LPC的分化。 TPA引起的变化是 通过免疫荧光和生物合成标记分析 SDS-PAGE。 我们试图确定是否有顺序 （分化连接？）识别的细胞表面结构的表达 通过单克隆抗体及其与细胞内的关系 LPC的标记，即TDT和CIGM。 结果表明 某些标记的获取/丢失可能与 淋巴样细胞前体中分化连接的事件。 （3）专业 目前重点放在检查细胞表面的功能上 分子GP40/BA-I，P24/BA-2和GP100/CARLA/BA-3对细胞进行介绍 他们在上面表达。 （4）特别感兴趣的是我们最近的 观察到gp100/calla在表面上大幅下降 严重热损伤患者的中性粒细胞。 由于中性粒细胞 从这些患者中表现出各种功能障碍（即在炎症中），我们 正在研究GP100/Calla在中性粒细胞功能中的可能作用。 （4） 正在进行的研究，以克隆编码GP40/BA-1，P24/BA-2和 GP/100/CACCA/BA-3使用小鼠L细胞转染和减法 杂交。 （磅）