Anti-inflammatory Cytokines in Atherosclerosis

描述(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。尽管干扰素-γ是典型的促炎细胞因子，但其在动脉粥样硬化中的作用尚不清楚。本研究的目的是通过检测IL-4和IL-10信号在炎症、巨噬细胞募集和病变形态中的作用，确定IL-4和IL-10信号在动脉粥样硬化形成过程中的作用。在目标1中，IL4和IL10受体缺陷的小鼠(IL4R和IL10R)将与易患动脉粥样硬化的LDLR-/-小鼠杂交，以评估这些细胞因子信号的丢失如何影响动脉粥样硬化。由于IL4R和IL10R在病变中的所有细胞类型上都有表达，因此对缺乏这些受体的小鼠的研究将确定细胞因子对白细胞的作用相对于病变中其他类型细胞的相对重要性。AIM 2中的研究旨在检测IL4和IL10在病变局部环境中的表达。利用巨噬细胞特异性启动子，IL4和IL10过表达的转基因小鼠将被产生并用作骨髓供体小鼠。由此产生的LDLR-/-嵌合小鼠的巨噬细胞在病变中过度表达IL4或IL10，这将为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的形成提供有价值的信息。目的3将研究IL4和IL10信号对巨噬细胞功能的影响，因为它们与动脉粥样硬化的形成有关。首先，将通过体外细胞黏附和体内细胞迁移试验来测试IL4和IL10介导巨噬细胞向病变募集的能力。此外，IL4和ILIO对细胞外基质(ECM)重塑的影响将首先通过测量这些细胞因子在巨噬细胞合成基质金属蛋白酶(MMPs)中的作用，然后通过检测没有IL4或IL10信号能力的动脉粥样硬化动物的病变形态来检验。对ECM蛋白沉积、基质金属蛋白酶表达和细胞浸润的检测将揭示这两种细胞因子在病变进展中的作用。这些研究将强调炎症在动脉粥样硬化中的重要性，并可能导致针对炎症的新治疗措施的开发，以减少动脉粥样硬化的发生。

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.