PHYSICIAN SCIENTIST AWARD

单核细胞刺激可产生白介素1(IL-1) 用许多不同的化合物完成的。然而，对这一问题的洞察却很少 对IL-1产生的负调控是可行的。因此，这 研究提案是一个分两个阶段的项目，旨在阐明对 这种单倍体的生产。 在第一阶段，我们将开始在单核细胞中鉴定IL-1的时间进程 蛋白质和基因的表达，2)不同刺激对基因表达的影响 IL-1基因的差异表达；3)IL-1的药理抑制 蛋白质或基因表达，4)花生四烯酸代谢产物在 IL-1蛋白或基因表达，5)淋巴细胞及淋巴细胞产物 参与调节IL-1蛋白或基因的表达。实验将会 在脂多糖或其他物质刺激的外周血单核细胞上进行 IL-1诱导剂。抑制实验将评估常用的 免疫调节药物(地塞米松和环孢素A)、环氧合酶 吲哚美辛和脂氧合酶对花生四烯酸的抑制作用 新陈代谢。分析IL-1蛋白表达将需要使用 单核细胞分离的标准组织培养技术，小鼠胸腺细胞 白介素1功能定量的生物测定法及Northern分析 使用针对IL-1mRNA的合成寡核苷酸的细胞RNA。 在第二阶段，免疫控制机制的应用将是 适用于肺泡巨噬细胞和结节病，我们假设 IL-1调节异常可能导致永久性残疾。我们会 在正常肺泡巨噬细胞和结节病中鉴定1)时间进程 对IL-1蛋白和基因表达的影响；2)不同刺激对大鼠脑内IL-1蛋白和基因表达的影响 IL-1基因的差异表达；3)IL-1的药理抑制 蛋白质和基因表达，4)正常淋巴细胞的作用，他们的 IL-1蛋白和基因的调节产物及其在结节病中的作用 表情。这些研究将通过使用标准 小鼠胸腺细胞分析和IL-1RNA的Northern分析。 随着对药理相关抑制剂的详细研究，新的 然后，治疗方案将变得可用于免疫调节 肺病，如结节病。对IL-1产生的调节可能是 在改变这些患者的预后和功能损害方面非常重要 疾病。

Monocyte stimulation resulting in interleukin-1 (IL-1) production can be accomplished with many different compounds. Yet little insight into the negative regulation of IL-1 production is available. Therefore, this research proposal is a two-phase project aimed at elucidating control of production of this monokine. In phase one, we will begin to identify in monocytes 1) time course of IL-1 protein and gene expression, 2) the effects of different stimuli on differential IL-1 gene expression, 3) pharmacological inhibition of IL-1 protein or gene expression, 4) the role of arachidonic acid metabolites in IL-1 protein or gene expression, 5) lymphocytes and lymphocyte products' place in regulation of IL-1 protein or gene expression. Experiments will be performed on peripheral blood monocytes stimulated with LPS or other IL-1 inducers. Inhibition experiments will evaluate commonly used immunomodulatory drugs (dexamethasone and cyclosporin A), cyclooxygenase (indomethacin) and lipoxygenase (U60) inhibition of arachidonic metabolism. Analysis of IL-1 protein expression will require use of standard tissue culture techniques for monocyte isolation, mouse thymocyte bioassays for IL-1 functional quantification and Northern analysis of total cellular RNA using a synthetic oligonucleotide directed against IL-1 mRNA. In phase two, application of immunological control mechanisms will be applied to alveolar macrophage and sarcoidosis, where we hypothesize that abnormal IL-1 regulation may lead to permanent disability. We will identify in normal alveolar macrophages and in sarcoidosis 1) time course of IL-1 protein and gene expression, 2) the effects of different stimuli on differential IL-1 gene expression, 3) pharmacological inhibition of IL-1 protein and gene expression, 4) the role of normal lymphocytes, their products and those in sarcoidosis in regulating IL-1 protein and gene expression. These studies will be accomplished through use of standard mouse thymocyte assays and Northern analysis of IL-1 RNA. With pharmacologically relevant inhibitors studied in detail, new therapeutic options will then become available for immunologically mediated lung disease, such as sarcoidosis. Regulation of IL-1 production may be very important in altering prognosis and functional impairment in these diseases.