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首页> 外文期刊>Clinical immunology: The official journal of the Clinical Immunology Society >Immunochemical characterization of purified human oxidized low-density lipoprotein antibodies.
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Immunochemical characterization of purified human oxidized low-density lipoprotein antibodies.

机译：纯化的人类氧化型低密度脂蛋白抗体的免疫化学表征。

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The goal of this study was to characterize the isotypes and reactivity of human autoantibodies to copper oxidized LDL (oxLDL). Forty-six purified oxLDL antibodies contained immunoglobulins of the three major isotypes, with a predominance of IgG, subclasses 1 and 3. These IgG isotypes are known to interact with FcRgammaI and to activate the complement system and thus are potentially able to activate macrophages and cause foam cell formation. The same purified antibodies were tested for cross-reactivity with malondialdehyde (MDA)-, glycated (Glyc)-, and native (n)LDL and cardiolipin. Absorption with oxLDL resulted in a decrease of reactivity of 77.2 +/- 4.7%. Absorption with MDA-LDL resulted in a wider range of reduction of reactivity values, ranging from 50 to 87%, possibly reflecting differences in the degree of MDA modification. Absorption with Glyc- and nLDL caused a minor decrease in the reactivity of antibodies to oxLDL (5.9 +/- 7.1 and 6.8 +/- 6. 4%, respectively), comparable to the reduction of reactivity (2.1 +/- 4.0%) measured after absorption with transferrin, an irrelevant protein used as a negative control. These results suggest that oxLDL antibodies recognize primarily MDA epitopes. To determine whether purified oxLDL antibodies also recognize other epitopes known to be generated during copper oxidation of LDL, such as 4-hydroxynonenal (HNE)- and N(varepsilon)(carboxymethyl)-lysine (CML), two additional sets of experiments were carried out. First, we monitored the formation of CML-, MDA-lysine, and HNE-lysine at different times during copper oxidation of two LDL pools. Both pools showed simultaneous increases in protein modification, as indicated by increasing fluorescence emission at 430 nm, and in immunoreactivity with oxLDL antibodies, coinciding closely with MDA modification of lysine groups. Second, we assessed whether the reactivity of oxLDL antibodies could be blocked by absorption with CML- or HNE-LDL. HNE-LDL did not react with isolated oxLDL antibodies. Highly modified CML-LDL (>90% of lysine residues modified) reduced the reactivity of oxLDL antibodies, but only by 25.5%. Finally, we investigated the possible cross-reactivity of oxLDL antibodies with cardiolipin. Seventeen purified oxLDL antibodies were used in this study, which showed that absorption with oxLDL or nLDL did not affect their reactivity with immobilized cardiolipin. Copyright 2000 Academic Press.
机译：这项研究的目的是表征人类自身抗体对铜氧化的LDL（oxLDL）的同种型和反应性。四十六种纯化的oxLDL抗体包含三种主要同种型的免疫球蛋白，主要为IgG，亚类1和3。这些IgG同种型已知与FcRgammaI相互作用并激活补体系统，因此有可能激活巨噬细胞并引起泡沫细胞形成。测试了相同的纯化抗体与丙二醛（MDA）-，糖化（Glyc）-和天然（n）LDL和心磷脂的交叉反应性。 oxLDL的吸收导致反应活性降低77.2 +/- 4.7％。 MDA-LDL的吸收导致反应性值的降低范围更广，范围从50％到87％，这可能反映了MDA修饰程度的差异。糖基和nLDL的吸收导致抗体对oxLDL的反应性轻微降低（分别为5.9 +/- 7.1和6.8 +/- 6. 4％），与反应性的降低（2.1 +/- 4.0％）相当用转铁蛋白（一种无关蛋白，用作阴性对照）吸收后测定。这些结果表明oxLDL抗体主要识别MDA表位。为了确定纯化的oxLDL抗体是否还识别LDL铜氧化过程中已知产生的其他表位，例如4-羟基壬烯醛（HNE）-和N（varepsilon）（羧甲基）-赖氨酸（CML），还进行了两套实验出来。首先，我们在两个LDL库的铜氧化过程中的不同时间监控CML-，MDA-赖氨酸和HNE-赖氨酸的形成。这两个库均显示出蛋白质修饰的同时增加（如430 nm处荧光发射的增加所指示）以及与oxLDL抗体的免疫反应性，这与赖氨酸基团的MDA修饰非常吻合。其次，我们评估了用CML-或HNE-LDL吸收是否可以阻断oxLDL抗体的反应性。 HNE-LDL不与分离的oxLDL抗体反应。高度修饰的CML-LDL（修饰了90％以上的赖氨酸残基）降低了oxLDL抗体的反应性，但仅降低了25.5％。最后，我们研究了oxLDL抗体与心磷脂的可能的交叉反应性。在该研究中使用了十七种纯化的oxLDL抗体，这表明oxLDL或nLDL的吸收不会影响它们与固定的心磷脂的反应性。版权所有2000学术出版社。

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