Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria.
J Lipid Res. 2012 Jul;53(7):1316-26. doi: 10.1194/jlr.M025445. Epub 2012 Apr 16.
Autoantibodies specific for malondialdehyde-modified LDL (MDA-LDL) represent potential biomarkers to predict cardiovascular risk. However, MDA-LDL is a high variability antigen with limited reproducibility. To identify peptide mimotopes of MDA-LDL, phage display libraries were screened with the MDA-LDL-specific IgM monoclonal Ab LRO4, and the specificity and antigenic properties of MDA mimotopes were assessed in vitro and in vivo. We identified one 12-mer linear (P1) and one 7-mer cyclic (P2) peptide carrying a consensus sequence, which bound specifically to murine and human anti-MDA monoclonal Abs. Furthermore, MDA mimotopes were found to mimic MDA epitopes on the surface of apoptotic cells. Immunization of mice with P2 resulted in the induction of MDA-LDL-specific Abs, which strongly immunostained human atherosclerotic lesions. We detected IgG and IgM autoAbs to both MDA mimotopes in sera of healthy subjects and patients with myocardial infarction and stable angina pectoris undergoing percutaneous coronary intervention, and the titers of autoAbs correlated significantly with respective Ab titers against MDA-LDL. In conclusion, we identified specific peptides that are immunological mimotopes of MDA. These mimotopes can serve as standardized and reproducible antigens that will be useful for diagnostic and therapeutic applications in cardiovascular disease.
自身抗体特异性的丙二醛修饰的 LDL (MDA-LDL) 代表潜在的生物标志物来预测心血管风险。然而,MDA-LDL 是一个高变异性抗原,其重现性有限。为了鉴定 MDA-LDL 的肽模拟物,用 MDA-LDL 特异性 IgM 单克隆抗体 LRO4 筛选噬菌体展示文库,并在体外和体内评估 MDA 模拟物的特异性和抗原特性。我们鉴定出一个 12 个氨基酸的线性 (P1) 和一个 7 个氨基酸的环化 (P2) 肽,其携带一个共识序列,特异性结合鼠和人抗 MDA 单克隆抗体。此外,MDA 模拟物被发现模拟凋亡细胞表面的 MDA 表位。用 P2 免疫小鼠可诱导 MDA-LDL 特异性 Abs,该 Abs 强烈免疫染色人动脉粥样硬化病变。我们在健康受试者和心肌梗死及稳定性心绞痛患者的血清中检测到针对这两种 MDA 模拟物的 IgG 和 IgM 自身抗体,并且自身抗体的滴度与针对 MDA-LDL 的相应 Ab 滴度显著相关。总之,我们鉴定出 MDA 的特异性肽模拟物。这些模拟物可以作为标准化和可重现的抗原,将有助于心血管疾病的诊断和治疗应用。