Cloning of monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E-deficient mice - Demonstration of epitopes of oxidized low density lipoprotein in human plasma
Cloning of monoclonal autoantibodies to epitopes of oxidized lipoproteins from apolipoprotein E-deficient mice - Demonstration of epitopes of oxidized low density lipoprotein in human plasma
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DOI：
10.1172/jci118853
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发表时间：
1996-08-01
影响因子：
15.9
通讯作者：
Witztum, JL
中科院分区：
文献类型：
--
作者：
Palinski, W;Horkko, S;Witztum, JL
当低密度脂蛋白经历脂质过氧化时，可能会形成许多反应产物，进而与脂质、脱辅基蛋白和蛋白质反应，产生免疫原性新表位。识别氧化低密度脂蛋白模型表位的自身抗体，例如丙二醛-赖氨酸，存在于血浆和人类动脉粥样硬化病变中。 由于 apo E 缺陷小鼠产生特别高滴度的此类自身抗体，我们使用它们的脾脏克隆了 13 种针对氧化 LDL 不同表位的单克隆抗体（“EO 抗体”），结合和竞争性 RIA 证明，即使在最初选择用于与相同筛选抗原结合的 EO 抗体之间，精细特异性也存在显着差异，例如，某些 EO 选择与丙二醛-LDL结合的抗体还识别铜氧化的LDL、丙烯醛-LDL或由花生四烯酸或亚油酸氧化产物修饰的LDL。 在apo E缺陷小鼠中发现了花生四烯酸或亚油酸氧化产物，表明各自的抗原在体内形成。在人循环LDL上也发现了一些EO单克隆抗体识别的表位，每种EO单克隆抗体对兔和人动脉粥样硬化病变进行了免疫染色，其中一些产生了不同的染色 晚期病变的模式。总之，这表明天然单克隆抗体识别脂蛋白氧化过程中形成的复杂结构的不同表位，或在不同病变部位独立形成的表位。我们的数据表明，体内发生了对氧化脂蛋白的大量不同表位的深刻免疫反应。 “天然”单克隆自身抗体的可用性应有助于识别诱导这种反应的特定表位。
Many reactive products may be formed when LDL undergoes lipid peroxidation, which in turn can react with lipids, apoproteins, and proteins, generating immunogenic neoepitopes, Autoantibodies recognizing model epitopes of oxidized low density lipoprotein, such as malondialdehyde-lysine, occur in plasma and in atherosclerotic lesions of humans and animals, Because apo E-deficient mice develop particularly high titers of such autoantibodies, we used their spleens to clone 13 monoclonal antibodies to various epitopes of oxidized LDL (''EO antibodies''), Binding and competitive RIAs demonstrated significant differences in fine specificity even between EO antibodies initially selected for binding to the same screening antigen, For example, some EO antibodies selected for binding to malondialdehyde-LDL also recognized copper oxidized LDL, acrolein-LDL, or LDL modified by arachidonic or linoleic acid oxidation products, Circulating IgG and IgM autoantibodies binding to copper-oxidized LDL, 4-hydroxynonenal-LDL, acrolein-LDL, and LDL modified with arachidonic or linoleic acid oxidation products were found in apo E-deficient mice, suggesting that the respective antigens are formed in vivo., Epitopes recognized by some of the EO monoclonal antibodies were also found on human circulating LDL, Each of the EO monoclonal antibodies immunostained rabbit and human atherosclerotic lesions, and some of them yielded distinct staining patterns in advanced lesions. Together, this suggests that the natural monoclonal antibodies recognize different epitopes of complex structures formed during oxidation of lipoproteins, or epitopes formed independently at different lesion sites, Our data demonstrate that a profound immunological response to a large number of different epitopes of oxidized lipoproteins occurs in vivo. The availability of ''natural'' monoclonal autoantibodies should facilitate the identification of specific epitopes inducing this response.