NK CELL GENESIS FROM HUMAN BONE MARROW PROGENITORS

来自人类骨髓祖细胞的 NK 细胞起源

• 批准号: 3200103
• 负责人: EVA LOTZOVA
• 金额: $ 15.83万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3200103
• 关键词: RNA splicing; bone marrow; cell cycle; cell differentiation; cell mediated lymphocytolysis test; clone cells; cytokine receptors; flow cytometry; hematopoietic stem cells; human tissue; interleukin 2; leukopoiesis; lymphokines; monoclonal antibody; natural killer cells; polymerase chain reaction; tissue /cell culture

Natural killer (NK) cells play an important function in antitumor and anti-microbial defense, in regulation of lymphoid and hematopoietic cells, in production of a variety of cytokines, and represent the dominant effector cells involved in lymphokine-activated killer cell (LAK) phenomenon. Despite the biological and clinical significance of NK cells, very little attention has been given to defining NK cell lineage and understanding the maturational events in NK cell development. Our proposal is aimed at bridging this gap. It is now possible to study NK cells lineage and development as a result of the recent advances in recombinant DNA technology, molecular approaches and availability of a plethora of monoclonal antibodies, cytokines and genetic probes. The specific aims of our investigations are to study: (1) Generation of NK cells from human bone marrow CD34+ progenitors and their subsets, including hematopoietic stem cells (HSC), using long-term bone marrow culture (LTBMC) systems; (2) Stepwise characterization of NK cell differentiation events in LTBMC and subsequent clonal analysis of NK cell progenitors; LTBMC will be analyzed for: morphology and for acquisition of cytoplasmic and cell surface antigens during maturation, using two and three-color flow cytometry; expression of different forms of NK cell tumor recognition structure (NK-TR), as determined by alternate mRNA splicing, using PCR technique; zeta subunit, using cell-permeabilization procedure; frequency of NK progenitors, using limiting dilution assay; an attempt will be made to prepare monoclonal antibodies against NK cell progenitors. (3) Events in development of NK cell cytotoxic and regulatory functions in LTBMC, including development of NK cell cytotoxic mechanism, acquisition of binding/lytic activity, granzyme and cytolysin systems, ability to produce cytokines and regulate hematopoiesis. This step-wise analysis of NK cell maturation, starting with the most primitive stem cell compartment, will allow us to "map", for the first time, NK differentiation pathway(s). This information will help to answer the questions of NK cell lineage and will be instrumental in determining the maturational pathways involved in acquisition of some of the NK cell biological functions. By "mapping" of normal NK cell matura- tion, it may be possible to detect divergent pathways of differentiation or asynchronous structure/function relationships in cancer patients, especially leukemia.

自然杀伤（NK）细胞在抗肿瘤和抗肿瘤治疗中发挥重要作用， 抗微生物防御，调节淋巴和造血 细胞，在生产各种细胞因子，并代表 淋巴因子激活的杀伤细胞中的优势效应细胞 (LAK)现象 尽管NK的生物学和临床意义 细胞，很少有人注意到定义NK细胞谱系 并了解NK细胞发育中的成熟事件。 我们 该提案旨在弥合这一差距。 现在可以研究NK 细胞谱系和发展的最新进展， 重组DNA技术、分子方法和 过多的单克隆抗体、细胞因子和遗传探针。 的 本研究的具体目的是研究：（1）NK的产生 来自人骨髓CD 34+祖细胞及其亚群的细胞， 包括造血干细胞（HSC）， （2）NK细胞的逐步表征 LTBMC中的分化事件和随后的NK细胞克隆分析 将分析LTBMC的形态学和采集 细胞质和细胞表面抗原在成熟过程中，使用两个和 三色流式细胞术;不同形式NK细胞的表达 肿瘤识别结构（NK-TR），由替代mRNA确定 剪接，使用PCR技术; ζ亚基，使用细胞透化 程序; NK祖细胞的频率，使用有限稀释法; 我们将尝试制备抗NK细胞的单克隆抗体 祖先(3)NK细胞细胞毒性和 LTBMC中的调节功能，包括NK细胞毒性的发展 机制，结合/裂解活性的获得，颗粒酶和溶细胞素 系统，产生细胞因子和调节造血的能力。 这 NK细胞成熟的逐步分析，从最 原始干细胞隔室，将使我们能够“地图”，第一次 时间、NK分化途径。 这些信息将有助于 回答NK细胞谱系的问题，并将有助于 确定成熟的途径参与收购的一些 NK细胞的生物学功能。 通过“映射”正常NK细胞成熟- 因此，有可能检测分化的不同途径， 或癌症患者中的异步结构/功能关系， 尤其是白血病