Anti-inflammatory Cytokines in Atherosclerosis

动脉粥样硬化中的抗炎细胞因子

• 批准号: 6984242
• 负责人: WILLIAM A BOISVERT
• 金额: $ 40.94万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6984242
• 关键词: atherosclerosis; biological signal transduction; cell adhesion; cell migration; cytokine receptors; digital imaging; enzyme linked immunosorbent assay; extracellular matrix; genetically modified animals; green fluorescent proteins; inflammation; interleukin 10; interleukin 4; intravital microscopy; laboratory mouse; leukocyte activation /transformation; macrophage; metalloendopeptidases; pathologic process; polymerase chain reaction; receptor expression; western blottings

DESCRIPTION (provided by applicant): There is little doubt that inflammation plays a major role in atherosclerosis. As mediators of inflammation, cytokines released from T lymphocytes have the ability to determine the inflammatory phenotype of the host. Although the prototypic pro-inflammatory cytokine, interferon-y has been implicated in atherosclerosis the role of the prototypic anti-inflammatory cytokines interleukin (IL) 4 and 10 is not as clear. The objective of this proposal is to determine the participation of IL4 and IL10 signaling in the atherogenic process by examining their effects on inflammation, macrophage recruitment and lesion morphology. In aim 1 mice deficient in IL4 and IL10 receptors (IL4R and IL10R) will be crossed onto the atherosclerosis-prone LDLR-/- mice to assess how the loss of signaling by these cytokine affects atherosclerosis. Because IL4R and IL10R are expressed on all cell types in the lesion, studies with mice lacking these receptors will determine the relative importance of the cytokine action on leukocytes versus other cell types in the lesion. Studies in aim 2 are designed to examine the expression of IL4 and IL10 in the local environment of the lesion. Using a macrophage-specific promoter, IL4 and IL10 overexpressing transgenic mice will be generated and used as bone marrow donor mice. The resulting chimeric LDLR-/- mice with macrophages overexpressing IL4 or IL10 in the lesion will provide valuable information on whether the presence of cytokines in the lesion environment can suppress inflammation and reduce lesion formation. Aim 3 will examine the influence of IL4 and IL10 signaling on well established functions of macrophages as they relate to atherogenesis. First, the ability of IL4 and IL10 to mediate macrophage recruitment to the lesions will be tested by in vitro cell adhesion as well as in vivo cell migration assays. In addition, IL4 and ILIO's influence on extracellular matrix (ECM) remodeling will be examined by first measuring the role of these cytokines in synthesis of matrix metalloproteinases (MMP) by the macrophages, and second, by examining the lesion morphology in atherosclerotic animals with no IL4 or IL10 signaling capability. Measurements of ECM protein deposition, MMP expression and cell infiltration will reveal the participation of these two cytokines in lesion progression. These studies will highlight the importance of inflammation in atherosclerosis and may lead to development of new therapeutic measures targeting inflammation to reduce atherogenesis.

描述（由申请人提供）：毫无疑问，炎症在动脉粥样硬化中起主要作用。作为炎症介质，T淋巴细胞释放的细胞因子具有决定宿主炎症表型的能力。虽然原型促炎细胞因子干扰素-γ与动脉粥样硬化有关，但原型抗炎细胞因子白细胞介素（IL）4和10的作用尚不清楚。本提案的目的是通过检查IL 4和IL 10信号对炎症、巨噬细胞募集和病变形态的影响来确定IL 4和IL 10信号在致动脉粥样硬化过程中的参与。在目的1中，将IL 4和IL 10受体（IL 4 R和IL 10 R）缺陷的小鼠与动脉粥样硬化易感性LDLR-/-小鼠杂交，以评估这些细胞因子的信号传导损失如何影响动脉粥样硬化。由于IL 4 R和IL 10 R在病变中的所有细胞类型上表达，因此对缺乏这些受体的小鼠的研究将确定细胞因子对病变中白细胞与其他细胞类型作用的相对重要性。目的2中的研究旨在检测IL 4和IL 10在病变局部环境中的表达。使用巨噬细胞特异性启动子，将产生IL 4和IL 10过表达转基因小鼠并用作骨髓供体小鼠。所得的在病变中具有过表达IL 4或IL 10的巨噬细胞的嵌合LDLR-/-小鼠将提供关于病变环境中细胞因子的存在是否可以抑制炎症和减少病变形成的有价值的信息。目的3将检查IL 4和IL 10信号传导对巨噬细胞的既定功能的影响，因为它们与动脉粥样硬化形成有关。首先，将通过体外细胞粘附以及体内细胞迁移测定来测试IL 4和IL 10介导巨噬细胞募集至病变的能力。此外，IL 4和IL 10对细胞外基质（ECM）重塑的影响将通过以下方式来检查：首先测量这些细胞因子在巨噬细胞合成基质金属蛋白酶（MMP）中的作用，其次，检查没有IL 4或IL 10信号传导能力的动脉粥样硬化动物中的病变形态。测量ECM蛋白沉积、MMP表达和细胞浸润将揭示这两种细胞因子在病变进展中的参与。这些研究将突出炎症在动脉粥样硬化中的重要性，并可能导致开发针对炎症的新治疗措施以减少动脉粥样硬化形成。