Malle E, Hazell L, Stocker R, Sattler W, Esterbauer H, Waeg G
Karl-Franzens University, Institutes of Medical Biochemistry, Graz, Austria.
Arterioscler Thromb Vasc Biol. 1995 Jul;15(7):982-9. doi: 10.1161/01.atv.15.7.982.
Oxidation of LDL is thought to contribute to the early stages of atherogenesis. Because myeloperoxidase is present in atherosclerotic lesions and can produce the strong oxidant hypochlorous acid (HOCl), which converts LDL into its high-uptake atherogenic form in vitro, we raised polyclonal and monoclonal antibodies (MoAbs) against HOCl-modified LDL (HOCl-LDL). Characterization of the polyclonal anti-human HOCl-LDL Abs showed that they cross-reacted strongly with 4-hydroxynonenal-, malondialdehyde-, and Cu(2+)-oxidized LDL. Similarly, polyclonal and some monoclonal Abs against aldehyde- and Cu(2+)-modified LDL cross-reacted with HOCl-LDL. In contrast to the polyclonal Abs, two selected hybridoma cell line supernatants containing MoAbs raised against HOCl-LDL (MoAb-A and MoAb-B) did not cross-react with either native LDL or aldehyde- or Cu(2+)-modified LDL. MoAb-A (clone 1B10A11, subtype IgG1 kappa) recognized an epitope that appeared to be specific for HOCl-LDL and depended on the tertiary structure of the (lipo)protein, as judged by a lack of cross-reactivity with HOCl-modified human and bovine serum albumin and a loss of reactivity associated with lipoprotein denaturation. MoAb-B (clone 2D10G9, subtype IgG2b kappa), on the other hand, gave identical titration curves with HOCl-LDL and HOCl-modified albumins, suggesting that this antibody recognized epitopes that are commonly generated on proteins that have been oxidized with HOCl. Thus, MoAb-A and MoAb-B may be useful tools for the investigation of a possible role for HOCl-mediated damage to (lipo)proteins in atherosclerosis and other inflammatory diseases.
低密度脂蛋白（LDL）的氧化被认为在动脉粥样硬化形成的早期阶段起作用。由于髓过氧化物酶存在于动脉粥样硬化病变中，并且能够产生强氧化剂次氯酸（HOCl），后者在体外可将LDL转化为其高摄取性的致动脉粥样硬化形式，因此我们制备了针对HOCl修饰的LDL（HOCl-LDL）的多克隆抗体和单克隆抗体（MoAbs）。多克隆抗人HOCl-LDL抗体的特性表明，它们与4-羟基壬烯醛、丙二醛和铜（2+）氧化的LDL发生强烈交叉反应。同样，针对醛和铜（2+）修饰的LDL的多克隆抗体和一些单克隆抗体也与HOCl-LDL发生交叉反应。与多克隆抗体不同，两种选定的含有针对HOCl-LDL产生的MoAbs的杂交瘤细胞系上清液（MoAb-A和MoAb-B）与天然LDL或醛或铜（2+）修饰的LDL均无交叉反应。MoAb-A（克隆1B10A11，亚型IgG1 κ）识别的表位似乎对HOCl-LDL具有特异性，并且依赖于（脂）蛋白的三级结构，这是通过与HOCl修饰的人和牛血清白蛋白缺乏交叉反应以及与脂蛋白变性相关的反应性丧失来判断的。另一方面，MoAb-B（克隆2D10G9，亚型IgG2b κ）与HOCl-LDL和HOCl修饰的白蛋白给出相同的滴定曲线，这表明该抗体识别在被HOCl氧化的蛋白质上通常产生的表位。因此，MoAb-A和MoAb-B可能是用于研究HOCl介导的（脂）蛋白损伤在动脉粥样硬化和其他炎症性疾病中可能作用的有用工具。
