REGULATION OF MACROPHAGE DEVELOPMENT

巨噬细胞发育的调节

• 批准号: 3087798
• 负责人: KAREN Simpson MOULTON
• 金额: $ 8.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-01-01 至 1995-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3087798
• 关键词: alveolar macrophages; atherosclerosis; cell differentiation; cytogenetics; developmental genetics; eicosanoid metabolism; gene expression; genetic regulation; genetic transcription; human tissue; immunofluorescence technique; laboratory rabbit; lipoxygenase; low density lipoprotein; macrophage; molecular cloning; neoplastic cell culture for noncancer research; nuclear runoff assay; phorbols; retinoids

White blood cell differentiation is controlled by hormone like factors that act to regulate complex patterns of gene expression. A number of leukemia cell lines from the granulocyte or monocyte/macrophage with agents such as retinoic acid and phorbol esters. Aspects of macrophage development will be studied by determining the effects of differentiating agents on the expression of genes in the stimulated cell lines. The 5- and 15- lipoxygenase (5-LO, 15-LO) enzymes are produced in mature macrophages and produce leukotrienes and lipoxins from arachadonic acid. The developmental regulation of the 5-LO and 15-LO genes will be investigated by looking for variable gene expression in HL-60, THP-1, and U-937 cells after treatment with differentiating agents. Levels of 5-LO and 15-LO mRNA will be measured using Northern blot hybridization and ribonuclease protection assays. Nuclear run on studies are proposed to study the rates of 5-LO and 15-LO transcription initiation. Screening strategies for cDNA libraries derived from mRNA in stimulated cells are proposed to identify a network of genes that are induced by retinoic acid and phorbol esters. The specific cDNA clones activated in differentiated cells will be initially characterized by sequence analysis and determining tissue specific expression. Antisera raised against peptides coded by these cDNAs will be used in immunofluorescence studies to determine subcellular localization. Selected clones will be further analyzed to identify their functional roles in macrophages involved in normal and pathologic conditions. The subset of clones expressed in monocyte derived foam cells isolated from atherosclerotic lesions will be assessed for the functions they might perform in the process of atherogenesis. Experimental manipulations that alter the expression of individual clones or interfere with their protein products will be tested for their effects on macrophage development, the expression of specific macrophage properties, and LDL metabolism in foam cells.

白细胞分化受激素样因子控制， 调节基因表达的复杂模式。 一些白血病 来自粒细胞或单核细胞/巨噬细胞的细胞系，例如 视黄酸和佛波酯。 巨噬细胞发育的各个方面将 通过确定分化剂对 受刺激的细胞系中基因的表达。 5- 和 15- 脂氧合酶（5-LO、15-LO）在成熟的巨噬细胞中产生， 从花生四烯酸产生白三烯和脂氧素。 发展性的 5-LO 和 15-LO 基因的调控将通过寻找 治疗后 HL-60、THP-1 和 U-937 细胞中的可变基因表达 与分化剂。 5-LO 和 15-LO mRNA 的水平将为 使用 Northern blot 杂交和核糖核酸酶保护进行测量 化验。 建议进行核运行研究以研究 5-LO 和 15-LO转录起始。 cDNA文库的筛选策略 提议从受刺激细胞中的 mRNA 衍生来识别网络 由视黄酸和佛波酯诱导的基因。 具体的 在分化细胞中激活的 cDNA 克隆最初将被 通过序列分析和确定组织特异性来表征 表达。 针对这些 cDNA 编码的肽产生的抗血清将 用于免疫荧光研究以确定亚细胞定位。 将进一步分析选定的克隆以确定其功能作用 参与正常和病理条件下的巨噬细胞。 的子集 在单核细胞来源的泡沫细胞中表达的克隆，分离自 将评估动脉粥样硬化病变的功能 在动脉粥样硬化形成过程中发挥作用。实验操作 改变单个克隆的表达或干扰其蛋白质 将测试产品对巨噬细胞发育的影响， 泡沫中特定巨噬细胞特性的表达和 LDL 代谢 细胞。