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.

白细胞分化受激素控制的控制，例如 作用以调节基因表达的复杂模式。 许多白血病 来自粒细胞或单核细胞/巨噬细胞的细胞系 视黄酸和佛波酯。 巨噬细胞发展的各个方面将 通过确定区分剂对 基因在刺激细胞系中的表达。 5-和15- 脂氧合酶（5-LO，15-LO）酶是在成熟的巨噬细胞中产生的 产生白细胞三烯和脂毒素从蛛网酸中产生。 发展 5-LO和15-LO基因的调节将通过寻找 治疗后HL-60，THP-1和U-937细胞中的可变基因表达 与不同的代理。 5-LO和15-LO mRNA的水平将是 使用Northern印迹杂交和核糖核酸酶保护测量 测定。 提出了研究核跑步，以研究5-LO的速率和 15-LO转录启动。 cDNA库的筛选策略 提出了源自刺激细胞中的mRNA，以确定 视黄酸和佛波酯诱导的基因。 具体 在分化细胞中激活的cDNA克隆最初将是 通过序列分析和确定组织特异性的特征 表达。 针对这些cDNA编码的肽提出的抗血清将是 用于免疫荧光研究以确定亚细胞定位。 选定的克隆将进一步分析以识别其功能角色 在涉及正常和病理状况的巨噬细胞中。 子集 在单核细胞衍生的泡沫细胞中表达的克隆从 将评估动脉粥样硬化病变的功能 在动脉粥样硬化过程中执行。实验操作 改变单个克隆的表达或干扰其蛋白质 产品将对它们对巨噬细胞发展的影响进行测试， 特定巨噬细胞特性的表达和泡沫中的LDL代谢 细胞。