Regulation of Chemokine Expression In Vivo

体内趋化因子表达的调节

• 批准号: 6640235
• 负责人: THOMAS A. HAMILTON
• 金额: $ 33.3万
• 依托单位: CLEVELAND CLINIC FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6640235
• 关键词: T lymphocyte; chemokine; gene expression; genetically modified animals; inflammation; laboratory mouse; protein biosynthesis; skin hypersensitivity; surgical wound; wound healing

DESCRIPTION (provided by the applicant): Inflammation following tissue injury is an essential process that protects the organism against challenge by environmental pathogens and orchestrates the restoration of tissue architecture and homeostasis. A major aspect of this activity is dependent upon the sequential infiltration of inflammatory neutrophils and monocytes that is regulated, in part, by the production of chemoattractant cytokines or chemokines. Studies conducted in cultured cell lines have demonstrated that many cell types are capable of producing chemokines in response to a broad spectrum of agents. Our laboratory and others have demonstrated that chemokine expression is regulated through cell type- and stimulus-specific alterations in transcription and mRNA stability that depend upon specific nucleotide regulatory sequences in DNA and mRNA present in individual chemokine genes. Recent findings from our laboratory show that chemokine expression in vivo occurs in a complex multiphase pattern that is likely to involve the participation of distinct cell types and stimuli in a temporally ordered fashion. We cannot, however, accurately predict which individual stimuli, cell types, and mechanisms are responsible for the pattern of chemokine gene expression seen in any given inflammatory setting. The overall goal of this proposal, therefore, is to determine how the regulatory mechanisms identified from our cell culture models translate into the complex pattern of cell type- and stimulus-specific chemokine gene expression observed in vivo. This will be accomplished with two experimental objectives. 1. We will use mouse models of surgical injury or contact hypersensitivity in the skin to determine (a) the primary responding cell populations in each phase of response and (b) the pro-inflammatory and/or anti-inflammatory mediators responsible for regulating expression of KC and MIP-2 chemokine genes. 2. We will use transgenic mice expressing reporter transgenes containing wild type or mutant versions of defined regulatory sequences to determine their importance in controlling cell type- and stimulus-specific chemokine gene transcription and mRNA stability in vivo.

描述（由申请人提供）：组织损伤后的炎症是保护生物体免受环境病原体攻击的基本过程，并协调组织结构和体内平衡的恢复。这种活性的一个主要方面是依赖于炎症中性粒细胞和单核细胞的顺序浸润，这在一定程度上是由趋化因子或趋化因子的产生所调节的。在培养细胞系中进行的研究表明，许多类型的细胞能够对广泛的药物产生趋化因子。我们的实验室和其他人已经证明，趋化因子的表达是通过细胞类型和刺激特异性的转录和mRNA稳定性改变来调节的，这取决于个体趋化因子基因中存在的DNA和mRNA中的特定核苷酸调节序列。我们实验室最近的研究结果表明，趋化因子在体内的表达以复杂的多相模式发生，可能涉及不同细胞类型和刺激在时间上有序的参与。然而，我们不能准确地预测在任何给定的炎症环境中，是哪种个体刺激、细胞类型和机制导致了趋化因子基因表达模式。因此，本提案的总体目标是确定从我们的细胞培养模型中确定的调节机制如何转化为体内观察到的细胞类型和刺激特异性趋化因子基因表达的复杂模式。这将通过两个实验目标来实现。1. 我们将使用手术损伤或皮肤接触性过敏的小鼠模型来确定(a)每个反应阶段的主要应答细胞群和(b)负责调节KC和MIP-2趋化因子基因表达的促炎和/或抗炎介质。2. 我们将使用转基因小鼠表达含有野生型或突变型定义的调控序列的报告基因，以确定它们在控制细胞类型和刺激特异性趋化因子基因转录和mRNA稳定性中的重要性。