INTERLEUKIN 13 STIMULATION OF HUMAN MONOCYTES

白细胞介素 13 对人类单核细胞的刺激

• 批准号: 6389312
• 负责人: Martha K Cathcart
• 金额: $ 25.93万
• 依托单位: CLEVELAND CLINIC FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-05-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6389312
• 关键词: atherosclerotic plaque; biological signal transduction; blood lipoprotein; cytokine receptors; enzyme activity; gel electrophoresis; human subject; immunologic assay /test; interleukin 13; leukocyte activation /transformation; linoleate; lipoxygenase; low density lipoprotein; mitogen activated protein kinase; monocyte; oxidized lipid; radiotracer; tissue /cell culture

Monocytes are a subset of white blood cells critical for normal immune function. They are important components of inflammatory responses and are believed to significantly contribute to atherosclerotic lesion pathology. In atherosclerotic lesions, monocyte/macrophages have been shown to express a lipid oxidizing enzyme, 15-lipoxygenase (15-LO), that produces potent inflammatory mediators. Unique among the products of this enzyme are oxidized linoleate which we have shown to be the predominant oxidized fatty acid in atherosclerotic lesions. Normal monocytes do not express 15-LO, but are induced to do so by exposure to the cytokine IL-13 and expression of this enzyme is believed to correlate with disease progression in animal models of atherosclerosis. The studies proposed in this application will explore the IL-13 triggered signal transduction pathways in primary human monocytes and relate them to the downstream expression of 15-LO. Studies in Aim I will characterize the IL-13 receptor complex in human monocytes. Next, in Aim 2, the association and activation of Jaks with these receptor components will be assessed and activated Stats will be identified. The activation of Ser/Thr kinases will be evaluated (in Aim 3), expanding on our preliminary data that MAP kinases are activated by IL-13. The role of Jaks in this activation process will be investigated. Studies in Aim 4 will explore the role of Ser/Thr phosphorylation in regulating Jak/Stat phosphorylation and function. The biologic relevance of each of the activated pathways to the expression of 15-lipoxygenase and the related enhancement of monocyte-mediated LDL oxidation is the topic to be examined in studies described in Aim 5. In this final aim, additional investigations will be conducted to determine the role of the identified, IL-13 responsive pathways to general monocyte function with the goal.of identifying signaling pathways that are selectively involved in 15-LO expression but not required for other essential monocyte functions such as phagocytosis and superoxide anion production. The novelty of these studies is derived from the unique approaches to be employed to specifically dissect the roles of these pathways in intact human monocytes by virtue of the fact that monocytes are very conducive to antisense oligodeoxyribonucleotide manipulation of specific protein expression. These studies will substantially enhance our understanding of the signaling mechanisms induced upon exposure of monocytes to IL-13 and may suggest specific and selective means for modulating this response.

单核细胞是对正常免疫功能至关重要的白细胞的子集。它们是炎症反应的重要组成部分，被认为会显着有助于动脉粥样硬化病变病理学。在动脉粥样硬化病变中，单核细胞/巨噬细胞已被证明可以表达脂质氧化酶，15-脂氧酶（15-LO），可产生有效的炎症介质。在该酶的产物中，独特的是氧化的Linoleate，我们证明是动脉粥样硬化病变中主要的氧化脂肪酸。正常的单核细胞不表达15-LO，而是通过暴露于细胞因子IL-13而引起的，并且该酶的表达与动脉粥样硬化动物模型中的疾病进展相关。本应用中提出的研究将探索原代人单核细胞中IL-13触发信号转导途径，并将其与15-LO的下游表达相关联。目标我将在人单核细胞中表征IL-13受体复合物。接下来，在AIM 2中，将评估JAKS与这些受体成分的关联和激活，并确定激活的统计数据。将评估SER/THR激酶的激活（在AIM 3中），并扩展了我们的初步数据，即IL-13激活了映射激酶。将研究JAKS在激活过程中的作用。 AIM 4中的研究将探讨Ser/THR磷酸化在调节JAK/Stat磷酸化和功能中的作用。每种活化途径与15脂氧合酶表达的生物学相关性以及单核细胞介导的LDL氧化的相关增强是在目标5中描述的研究中要研究的主题。在此最终目标中，将进行其他调查，以确定鉴定出识别的识别的识别的途径，以确定识别的途径，该途径与该目标相关的作用，以确定识别该目标的作用，该途径识别出识别的途径，该途径与该目标相关。其他必要的单核细胞功能（例如吞噬作用和超氧化阴离子产生）所需的所需。这些研究的新颖性来自用于特异性地剖析这些途径在完整人单核细胞中的作用的独特方法，这是因为单核细胞非常有利于对特定蛋白质表达的反义寡脱氧核苷酸操纵。这些研究将大大增强我们对单核细胞暴露于IL-13诱导的信号传导机制的理解，并可能提出调节此响应的特定和选择性手段。