Up-regulation of adhesion molecule expression in glomerular endothelial cells by anti-myeloperoxidase antibody
Up-regulation of adhesion molecule expression in glomerular endothelial cells by anti-myeloperoxidase antibody
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DOI：
10.1093/ndt/gfl555
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发表时间：
2007-01-01
影响因子：
6.1
通讯作者：
Suzuki, Kazuo
中科院分区：
文献类型：
--
作者：
Nagao, Tomokazu;Matsumura, Mimiko;Suzuki, Kazuo
背景抗中性粒细胞髓过氧化物酶胞浆抗体（MPO-ANCA）与少免疫性新月体肾炎有关。它刺激致敏的中性粒细胞粘附于肾小球内皮细胞（GECs），从而释放活性氧和其他有毒物质，并最终损害GECs。虽然，MPO-ANCA的致病作用尚未完全了解，我们假设MPO-ANCA通过增加粘附分子的表达来调节GEC功能。在小鼠GEC（mGEC）中评价多克隆兔抗重组小鼠MPO抗体（抗rmMPO IgG）对粘附分子表达的影响。采用实时荧光定量逆转录聚合酶链反应（RT-PCR）和细胞间粘附分子-1（ICAM-1）细胞ELISA法检测mGEC与抗rmMPO IgG或同种型对照孵育后ICAM-1的表达。实时RT-PCR分析显示，用100 μ g/ml抗rmMPO IgG处理使ICAM-1、血管细胞粘附分子-1和E-选择素的mRNA表达分别增加约12.5、7.5和10.5倍。ICAM-1细胞ELISA也证实ICAM-1表达增加。ICAM-1表达的增强是由抗rmMPO IgG的抗原特异性介导的。此外，mGEC中有几种蛋白质可与抗rmMPO IgG特异性免疫沉淀。这些结果表明，抗MPO抗体不仅激活中性粒细胞，而且还激活GEC，表明抗rmMPO IgG诱导的GEC的直接激活有助于中性粒细胞粘附于GEC，从而在少免疫性肾小球肾炎的开始和进展中增加肾小球中性粒细胞浸润。
Background. Anti-neutrophil cytoplasmic antibody directed against myeloperoxidase (MPO-ANCA) has been implicated in pauci-immune crescentic glomerulonephritis. It stimulates primed neutrophils to adhere to glomerular endothelial cells (GECs), thereby releasing reactive oxygen and other toxic substances and ultimately damaging the GECs. Though, a pathogenic role for MPO-ANCA is not fully understood, we hypothesized that MPO-ANCA modulates GEC functions by the increases in expression of adhesion molecules.Methods. A polyclonal rabbit anti-recombinant mouse MPO antibody (anti-rmMPO IgG) was evaluated in mouse GEC (mGEC) for its effect on adhesion molecule expression. The primary culture of mGEC was incubated with anti-rmMPO IgG or isotype control and the expression of intercellular adhesion molecules-1 (ICAM-1) was evaluated by real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis and ICAM-1 cell ELISA.Results. The real-time RT-PCR analysis showed that a treatment with 100 mu g/ml anti-rmMPO IgG increased the expression of mRNAs for ICAM-1, vascular cell adhesion molecule-1 and E-selectin by approximately 12.5, 7.5 and 10.5-fold, respectively. ICAM-1 cell ELISA also substantiated increased expression of ICAM-1. This enhancement of ICAM-1 expression was mediated by the antigen specificity of anti-rmMPO IgG. In addition, there were several proteins in mGEC specifically immunoprecipitated with anti-rmMPO IgG.Conclusions. These results showed that anti-MPO antibody activates not only neutrophils, but also GEC, indicating that anti-rmMPO IgG-induced direct activation of GEC contributes to neutrophil adhesion to GEC, thereby increasing glomerular neutrophil infiltration in initiation and progression of pauci-immune glomerulonephritis.