Dynamic immune cell accumulation during flow-induced atherogenesis in mouse carotid artery: an expanded flow cytometry method.
Dynamic immune cell accumulation during flow-induced atherogenesis in mouse carotid artery: an expanded flow cytometry method.
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DOI：
10.1161/atvbaha.111.242180
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发表时间：
2012-03
期刊：
影响因子：
--
通讯作者：
Jo H
中科院分区：
文献类型：
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作者：
Alberts-Grill N;Rezvan A;Son DJ;Qiu H;Kim CW;Kemp ML;Weyand CM;Jo H
炎症在动脉粥样硬化中起核心作用。然而，动脉粥样硬化形成过程中动脉壁中白细胞的组成和数量的详细变化尚未完全了解，部分原因是缺乏合适的方法和动物模型。我们开发了一种10-荧光染料，13-参数流式细胞术的方法来定量7个主要的白细胞亚群在一个单一的消化动脉壁样本。ApoE−/−小鼠接受左颈动脉（LCA）部分结扎，并喂食高脂饲料4至28天。单核细胞/巨噬细胞、树突状细胞、粒细胞、NK细胞和CD 4 T细胞早在4d就显著浸润LCA。单核/巨噬细胞和树突状细胞在7 - 14天之间减少，而T细胞数量保持稳定。白细胞数在第7天达到峰值，在第14天形成动脉粥样硬化之前。B细胞于14 d进入LCA。对照组右侧颈动脉和假结扎组LCAs未见明显浸润。PCR和ELISA阵列显示，促炎细胞因子和趋化因子的表达分别在结扎后7天和14天达到峰值。这是第一次定量描述白细胞数量和组成的寿命小鼠动脉粥样硬化。这些结果表明，在动脉粥样硬化的发生和发展过程中，扰动的血流诱导动脉壁中快速和动态的白细胞积聚。
Inflammation plays a central role in atherosclerosis. However, the detailed changes in the composition and quantity of leukocytes in the arterial wall during atherogenesis are not fully understood due in part to the lack of suitable methods and animal models. We developed a 10-fluorochrome, 13-parameter flow cytometry method to quantitate 7 major leukocyte subsets in a single digested arterial wall sample. ApoE−/− mice underwent left carotid artery (LCA) partial ligation and fed high-fat diet for 4 to 28 days. Monocyte/macrophages, dendritic cells, granulocytes, NK cells, and CD4 T-cells significantly infiltrated the LCA as early as 4d. Monocyte/macrophages and dendritic cells decreased between 7d and 14d, while T-cell numbers remained steady. Leukocyte numbers peaked at 7d, preceding atheroma formation at 14d. B-cells entered LCA by 14d. Control right carotid and sham-ligated LCAs showed no significant infiltrates. PCR and ELISA arrays showed that expression of pro-inflammatory cytokines and chemokines peaked at 7 and 14-days post-ligation, respectively. This is the first quantitative description of leukocyte number and composition over the life span of murine atherosclerosis. These results show that disturbed flow induces rapid and dynamic leukocyte accumulation in the arterial wall during the initiation and progression of atherosclerosis.