Slot M C, Theunissen R, van Paassen P, Damoiseaux J G M C, Tervaert J W Cohen
Department of Clinical and Experimental Immunology, University Hospital Maastricht, Maastricht, The Netherlands.
Clin Exp Immunol. 2007 Aug;149(2):257-64. doi: 10.1111/j.1365-2249.2007.03420.x. Epub 2007 May 22.
Many patients surviving vasculitis are prone to accelerated atherosclerosis and often have enhanced levels of antibodies to oxidized low-density lipoprotein (oxLDL). To measure anti-oxLDL antibodies, oxidation of LDL is achieved with copper (Cu) or malondialdehyde (MDA). Because, in vivo, LDL may be oxidized with myeloperoxidase (MPO) or its product hypochlorite, we measured anti-hypochlorite LDL antibodies in patients with vasculitis, haemodialysis patients and healthy controls. A newly developed enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies to oxLDL as modified by hypochlorite. Results are compared with data obtained by standard LDL oxidation using MDA-LDL or Cu-LDL as substrate. Results were compared between anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) patients (n = 93), haemodialysis (HD) patients (n = 59) and healthy controls (HC; n = 43). Furthermore, patients with MPO-ANCA-associated vasculitis (n = 47) were compared with patients with proteinase 3 (PR3)-ANCA associated vasculitis (n = 46). Optimal cut-off points were determined by receiver operator characteristic (ROC) curve analysis. Anti-oxLDL antibodies are enhanced in AAV patients (MDA-LDL and hypochlorite-LDL) and in HD patients (hypochlorite-LDL), when compared to HC. Furthermore, patients with MPO-ANCA-associated vasculitis had higher levels of antibodies to hypochlorite-LDL than patients with PR3-ANCA-associated vasculitis. Our newly developed assay, in which hypochlorite-LDL is used as substrate, seems a more sensitive assay than traditional assays to measure oxLDL antibodies. Furthermore, our results suggest that enhanced MPO-mediated LDL oxidation occurs in patients with MPO-ANCA.
许多血管炎幸存者易患动脉粥样硬化加速,且往往具有较高水平的氧化型低密度脂蛋白(oxLDL)抗体。为了检测抗oxLDL抗体,用铜(Cu)或丙二醛(MDA)使低密度脂蛋白(LDL)氧化。由于在体内,LDL可能被髓过氧化物酶(MPO)或其产物次氯酸盐氧化,我们检测了血管炎患者、血液透析患者和健康对照者的抗次氯酸盐LDL抗体。使用新开发的酶联免疫吸附测定(ELISA)来检测经次氯酸盐修饰的oxLDL抗体。将结果与使用MDA-LDL或Cu-LDL作为底物的标准LDL氧化所获得的数据进行比较。在抗中性粒细胞胞浆抗体(ANCA)相关血管炎(AAV)患者(n = 93)、血液透析(HD)患者(n = 59)和健康对照者(HC;n = 43)之间比较结果。此外,将MPO-ANCA相关血管炎患者(n = 47)与蛋白酶3(PR3)-ANCA相关血管炎患者(n = 46)进行比较。通过受试者操作特征(ROC)曲线分析确定最佳截断点。与HC相比,AAV患者(MDA-LDL和次氯酸盐-LDL)和HD患者(次氯酸盐-LDL)中的抗oxLDL抗体增加。此外,MPO-ANCA相关血管炎患者的抗次氯酸盐-LDL抗体水平高于PR3-ANCA相关血管炎患者。我们新开发的以次氯酸盐-LDL作为底物的检测方法,似乎比传统检测方法更灵敏地检测oxLDL抗体。此外,我们的结果表明,MPO-ANCA患者中发生了MPO介导的LDL氧化增强。