ANTISERA AND MONOCLONAL-ANTIBODIES SPECIFIC FOR EPITOPES GENERATED DURING OXIDATIVE MODIFICATION OF LOW-DENSITY-LIPOPROTEIN
ANTISERA AND MONOCLONAL-ANTIBODIES SPECIFIC FOR EPITOPES GENERATED DURING OXIDATIVE MODIFICATION OF LOW-DENSITY-LIPOPROTEIN
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DOI：
10.1161/01.atv.10.3.325
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发表时间：
1990-05-01
期刊：
影响因子：
--
通讯作者：
WITZTUM, JL
中科院分区：
文献类型：
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作者：
PALINSKI, W;YLAHERTTUALA, S;WITZTUM, JL
越来越多的证据表明，低密度脂蛋白（LDL）必须进行修饰，以诱导泡沫细胞的形成。一种这样的修饰，LDL的氧化，产生许多能够与脱辅基蛋白B的赖氨酸残基缀合的氧化脂肪酸的高反应性短链长度的双链片段。通过免疫动物与同源的丙二醛修饰的低密度脂蛋白（MDA-LDL），4-羟基壬烯醛-LDL（4-HNE-LDL），和Cu++氧化的LDL，我们开发了多价和单克隆抗体对三个表位中发现的氧化修饰的LDL。本文描述了一种特异于MDA-赖氨酸的抗血清和单克隆抗体（分别为MAL-2和MDA-2）以及一种特异于4-HNE-赖氨酸的抗血清和单克隆抗体（分别为HNE-6和NA-59）。此外，针对在LDL的Cu++氧化过程中产生的尚未确定的表位开发了单克隆抗体（OLF 4 - 3C 10）。用这些抗体，我们证明了MDA-赖氨酸和4-HNE-赖氨酸加合物在铜诱导的LDL体外氧化过程中在载脂蛋白B上形成。这些抗体用于氧化脂蛋白在进行性严重动脉粥样硬化病变中的免疫细胞化学显示的应用在配套文章中描述。这些抗体应证明在研究氧化修饰的脂蛋白以及其他氧化修饰的蛋白质在动脉粥样硬化形成中的作用有用。
Increasing evidence indicates that low density lipoprotein (LDL) has to be modified to induce foam cell formation. One such modification, oxidation of LDL, generates a number of highly reactive short chain-length aldehydic fragments of oxidized fatty acids capable of conjugating with lysine residues of apoprotein B. By immunizing animals with homologous malondialdehyde-modified LDL (MDA-LDL), 4-hydroxynonenal-LDL (4-HNE-LDL), and Cu++-oxidized LDL, we developed polyvalent and monoclonal antibodies against three epitopes found in oxidatively modified LDL. The present article characterizes an antiserum and monoclonal antibody (MAL-2 and MDA2, respectively) specific for MDA-lysine, and an antiserum and monoclonal antibody (HNE-6 and NA59, respectively) specific for 4-HNE-lysine. In addition, a monoclonal antibody (OLF4-3C10) was developed against an as yet undefined epitope generated during Cu++ oxidation of LDL. With these antibodies, we demonstrated that MDA-lysine and 4-HNE-lysine adducts develop on apo-lipoprotein B during copper-induced oxidation of LDL in vitro. The application of these antibodies for immunocytochemical demonstration of oxidized lipoproteins in atherosclerotic lesions of progressive severity is described in the companion article. These antibodies should prove useful in studying the role of oxidatively modified lipoproteins as well as other oxidatively modified proteins in atherogenesis.