Ablation of the Ccr2 Gene Exacerbates Polyarthritis in Interleukin-1 Receptor Antagonist-Deficient Mice
Ablation of the Ccr2 Gene Exacerbates Polyarthritis in Interleukin-1 Receptor Antagonist-Deficient Mice
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DOI：
10.1002/art.30106
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发表时间：
2011-01-01
影响因子：
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通讯作者：
Mukaida, Naofumi
中科院分区：
文献类型：
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作者：
Fujii, Hiroshi;Baba, Tomohisa;Mukaida, Naofumi
Objective.类风湿关节炎（RA）的发病机制涉及细胞因子和趋化因子。鉴于关节内巨噬细胞浸润在RA中的作用，本研究旨在探讨巨噬细胞大量表达的趋化因子受体CCR 2在IL 1 Rn缺陷小鼠（RA小鼠模型）中的致病作用。对IL 1 rn缺陷和IL 1 rn和Ccr 2双缺陷小鼠进行关节炎的临床评估和组织学检查。骨密度测量采用计算机断层扫描。免疫组化和流式细胞术分析关节浸润细胞的类型。抗酒石酸酸性磷酸酶染色后定量关节中的破骨细胞。细胞因子和趋化因子水平用酶联免疫吸附试验和多重悬浮阵列测定。用免疫荧光双标记法检测趋化因子和破骨细胞生成因子的表达。将抗小鼠CXCR 2抗体注射到Il 1 rn和Ccr 2双缺陷小鼠体内进行阻断实验。Ccr 2基因的消融实际上加剧了关节炎和关节内破骨细胞的生成，而它增强了IL 1 Rn缺陷小鼠关节内中性粒细胞的积聚，但没有增强巨噬细胞的积聚。浸润的中性粒细胞表达破骨细胞生成因子RANKL和ADAM-8，从而增强Il 1 rn和Ccr 2双缺陷小鼠关节内破骨细胞生成。此外，双缺陷小鼠表现出增强的嗜酸性趋化因子角质形成细胞趋化因子和巨噬细胞炎性蛋白2（MIP-2）的表达，与IL-1 rn缺陷小鼠相比。最后，针对CXCR 2（角质形成细胞趋化因子和MIP-2的受体）的中和抗体显著减轻了IL 1 rn和Ccr 2双缺陷小鼠的关节炎。我们的研究结果表明，CCR 2介导的信号可以通过负调节中性粒细胞浸润来调节IL 1 rn缺陷小鼠的关节炎。
Objective. The pathogenesis of rheumatoid arthritis (RA) involves cytokines and chemokines. Given the role of intraarticular macrophage infiltration in RA, this study was undertaken to address the pathogenic role of CCR2, a chemokine receptor that is abundantly expressed by macrophages, in Il1rn-deficient mice, a mouse model of RA.Methods. Il1rn-deficient and Il1rn and Ccr2-double-deficient mice were subjected to clinical assessment of arthritis and histologic examination. Bone mineral density was measured with computed tomography. The types of cells infiltrating joints were determined by immunohistochemical analysis and flow cytometric analysis. Osteoclasts in joints were quantified after tartrate-resistant acid phosphatase staining. Cytokine and chemokine levels were measured by enzyme-linked immunosorbent assay and multiplex suspension array assay. The expression patterns of chemokines and osteoclastogenic factors were determined by double-color immunofluorescence analysis. Anti-mouse CXCR2 antibody was injected into Il1rn and Ccr2-double-deficient mice for blocking experiments.Results. Ablation of the Ccr2 gene actually exacerbated arthritis and intraarticular osteoclastogenesis, while it enhanced intraarticular neutrophil but not macrophage accumulation in Il1rn-deficient mice. Infiltrated neutrophils expressed the osteoclastogenic factors RANKL and ADAM-8, thereby augmenting intraarticular osteoclastogenesis in Il1rn and Ccr2-double-deficient mice. Moreover, the double-deficient mice exhibited enhanced expression of the neutrophilic chemokines keratinocyte chemoattractant and macrophage inflammatory protein 2 (MIP-2), compared with Il1rn-deficient mice. Finally, neutralizing antibodies to CXCR2, the receptor for keratinocyte chemoattractant and MIP-2, dramatically attenuated arthritis in Il1rn and Ccr2-double-deficient mice.Conclusion. Our findings indicate that CCR2-mediated signals can modulate arthritis in Il1rn-deficient mice by negatively regulating neutrophil infiltration.