REGULATION OF MACROPHAGE DEVELOPMENT

白色血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， 视黄酸和佛波醇酯。 巨噬细胞发育的各个方面 通过确定分化剂对 基因在刺激的细胞系中的表达。 5和15- 脂氧合酶（5-LO，15-LO）在成熟巨噬细胞中产生， 从花生四烯酸中产生白三烯和脂氧素。 发育 5-LO和15-LO基因的调控将通过寻找 处理后HL-60、THP-1和U-937细胞中的可变基因表达 差异化代理。 5-LO和15-LO mRNA的水平将被 使用北方印迹杂交和核糖核酸酶保护进行测量 测定。 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 从刺激的细胞中的mRNA衍生的，提出了确定一个网络， 由视黄酸和佛波酯诱导的基因。 具体 在分化的细胞中激活的cDNA克隆最初将被激活。 通过序列分析和确定组织特异性 表情 针对由这些cDNA编码的肽产生的抗血清将被免疫。 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，以确定其功能作用 在正常和病理条件下参与的巨噬细胞中。 的子集 克隆表达于分离的单核细胞衍生的泡沫细胞， 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它们的蛋白质 产品将测试其对巨噬细胞发育的影响， 特定巨噬细胞特性的表达和泡沫中的LDL代谢 细胞

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.