Department of Medicine, University of California, San Diego, La Jolla, CA 92093.
Cardiovascular Research Center, Department of Medicine, University of Virginia, Charlottesville, VA 22908.
J Lipid Res. 2019 Feb;60(2):436-445. doi: 10.1194/jlr.D090852. Epub 2018 Dec 18.
Atherosclerosis is associated with increased lipid peroxidation, leading to generation of multiple oxidation-specific epitopes (OSEs), contributing to the pathogenesis of atherosclerosis and its clinical manifestation. Oxidized cholesteryl esters (OxCEs) are a major class of OSEs found in human plasma and atherosclerotic tissue. To evaluate OxCEs as a candidate biomarker, we generated a novel mouse monoclonal Ab (mAb) specific to an OxCE modification of proteins. The mAb AG23 (IgG1) was raised in C57BL6 mice immunized with OxCE-modified keyhole limpet hemocyanin, and hybridomas were screened against OxCE-modified BSA. This method ensures mAb specificity to the OxCE modification, independent of a carrier protein. AG23 specifically stained human carotid artery atherosclerotic lesions. An ELISA method, with AG23 as a capture and either anti-apoAI or anti-apoB-100 as the detection Abs, was developed to assay apoAI and apoB-100 lipoproteins that have one or more OxCE epitopes. OxCE-apoA or OxCE-apoB did not correlate with the well-established oxidized phospholipid-apoB biomarker. In a cohort of subjects treated with atorvastatin, OxCE-apoA was significantly lower than in the placebo group, independent of the apoAI levels. These results suggest the potential diagnostic utility of a new biomarker assay to measure OxCE-modified lipoproteins in patients with CVD.
动脉粥样硬化与脂质过氧化增加有关，导致产生多种氧化特异性表位（OSEs），这有助于动脉粥样硬化的发病机制及其临床表现。氧化胆固醇酯（OxCEs）是人类血浆和动脉粥样硬化组织中主要的 OSE 类别之一。为了评估 OxCEs 作为候选生物标志物的能力，我们生成了一种针对蛋白质 OxCE 修饰的新型小鼠单克隆抗体（mAb）。该 mAb AG23（IgG1）是在 C57BL6 小鼠中通过免疫 OxCE 修饰的血蓝蛋白产生的，并用 OxCE 修饰的 BSA 筛选杂交瘤。这种方法确保了 mAb 对 OxCE 修饰的特异性，与载体蛋白无关。AG23 特异性染色人类颈动脉粥样硬化病变。开发了一种 ELISA 方法，使用 AG23 作为捕获抗体，使用抗 apoAI 或抗 apoB-100 作为检测抗体，以检测具有一个或多个 OxCE 表位的 apoAI 和 apoB-100 脂蛋白。OxCE-apoA 或 OxCE-apoB 与已建立的氧化磷脂-apoB 生物标志物没有相关性。在接受阿托伐他汀治疗的患者队列中，OxCE-apoA 明显低于安慰剂组，而与 apoAI 水平无关。这些结果表明，在 CVD 患者中测量 OxCE 修饰的脂蛋白的新生物标志物测定具有潜在的诊断效用。
