Expression and characterization of the chemokine receptors CCR2 and CCR5 in mice
Expression and characterization of the chemokine receptors CCR2 and CCR5 in mice
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DOI：
10.4049/jimmunol.166.7.4697
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发表时间：
2001-04-01
影响因子：
4.4
通讯作者：
Schlöndorff, D
中科院分区：
文献类型：
--
作者：
Mack, M;Cihak, J;Schlöndorff, D
趋化因子受体CCR2和CCR5在单核/巨噬细胞和T细胞的募集中发挥重要作用。为了更好地理解这两种受体在小鼠炎症性疾病模型中的作用，并在将这些数据与人类相关联时识别潜在问题，我们已经生成了针对小鼠CCR2和CCR5的单克隆抗体。在小鼠中，CCR2在单核细胞和2-15%的T细胞上均匀表达，与人类的表达模式非常相似。与人类相比，小鼠NK细胞CCR5高度阳性。此外，CCR5在3-10%的CD4和10-40%的cd8阳性T细胞上表达，在单核细胞上检测较弱。使用免疫复合物肾炎模型，我们检查了炎症对趋化因子受体表达的影响，发现炎症肾脏中CCR5（+）和CCR2(+) T细胞富集10倍。通过配体诱导CCR2和CCR5在原代白细胞上的内化，检测了各种趋化因子的活性和单克隆抗体的拮抗特性。Ab MC-21（抗ccr2）使小鼠单核细胞趋化蛋白1的活性降低了95%，而Ab MC-68（抗ccr5）阻断了99%以上的巨噬细胞炎症蛋白1 α和RANTES活性。MC-21和MC-68有效阻断了配体与CCR2和CCR5的结合，IC50分别为0.09和0.6-1.0 mug/ml。与这些体外数据有很好的相关性，MC-21几乎完全阻止了巯基乙酸盐诱导的腹膜炎中单核细胞的涌入。因此，这两种抗体都是进一步研究CCR2和CCR5在小鼠疾病模型中的作用的有用试剂。
The chemokine receptors CCR2 and CCR5 play important roles in the recruitment of monocytes/macrophages and T cells. To better understand the role of both receptors in murine models of inflammatory diseases and to recognize potential problems when correlating these data to humans, we have generated mAbs against murine CCR2 and CCR5. In mice CCR2 is homogeneously expressed on monocytes and on 2-15% of T cells, closely resembling the expression pattern in humans. In contrast to humans, murine NK cells are highly CCR5 positive. In addition, CCR5 is expressed on 3-10% of CD4 and 10-40% of CD8-positive T cells and is weakly detectable on monocytes. Using a model of immune complex nephritis, we examined the effects of inflammation on chemokine receptor expression and found a 10-fold enrichment of CCR5(+) and CCR2(+) T cells in the inflamed kidneys. The activity of various chemokines and the antagonistic properties of the mAbs were measured by ligand-induced internalization of CCR2 and CCR5 on primary leukocytes. The Ab MC-21 (anti-CCR2) reduced the activity of murine monocyte chemotactic protein 1 by 95%, whereas the Ab MC-68 (anti-CCR5) blocked over 99% of the macrophage-inflammatory protein 1 alpha and RANTES activity. MC-21 and MC-68 efficiently blocked the ligand binding to CCR2 and CCR5 with an IC50 of 0.09 and 0.6-1.0 mug/ml, respectively. In good correlation to these in vitro data, MC-21 almost completely prevented the influx of monocytes in thioglycollate-induced peritonitis. Therefore, both Abs appear as useful reagents to further study the role of CCR2 and CCR5 in murine disease models.