Analysis of Apolipoprotein H in Lupus

资料(申请人提供)：罹患冠心病的风险 在系统性红斑狼疮(SLE)中，女性的发病率是女性的50倍 普通民众。传统的风险因素不足以 解释系统性红斑狼疮患者的早发冠心病。相比之下，患病率约为1%-5% 在普通美国白人人群中的抗磷脂抗体(APA)， 大约50%的SLE患者APA呈阳性。ApoH是一名校长 自身免疫性疾病患者产生APA的自身抗原。 载脂蛋白H抑制氧化型低密度脂蛋白的体外摄取 通过巨噬细胞，但在APA的存在下，它促进oxLDL的ihflow进入 巨噬细胞。由于oxLDL在巨噬细胞中的积聚被认为是 启动动脉粥样硬化的过程，这些发现表明载脂蛋白H介导的 自身免疫性疾病患者的免疫反应，如SLE，可能导致 动脉硬化。在这次更新中，我们建议审查《行政程序法》的联合作用， 氧化低密度脂蛋白抗体(抗氧化低密度脂蛋白)和载脂蛋白基因变异 作为本建议的一部分被发现)与#年冠心病的发生有关 SUE患者和非SLE患者。我们的假设是APA阳性的个体 和/或抗ox低密度脂蛋白容易发生过早冠心病，这种易感性是 被APOH基因中常见的遗传变异所修饰。该计划的目标 通过实现这五个目标来实现学习。目标1)确定和 描述自然发生的所有外显子、内含子和 变性高效液相色谱聚合酶链式反应检测APOH基因3‘端序列 系统性红斑狼疮和非系统性红斑狼疮冠心病患者以及非洲黑人的分析和DNA测序 APA检测呈阳性。目的2)确定APA的患病率及其相关性 (抗载脂蛋白、抗心磷脂、狼疮抗凝剂)和抗氧化型低密度脂蛋白 样本来自SLE患者和对照组。目标3)确定关系 APOH基因变异(在目标1中产生的数据)与 APA和抗oxLDL(在AIM 2中生成的数据)。目标4)检查关系 APOH基因变异(在目标1中产生的数据)与 系统性红斑狼疮和冠心病患者的亚临床心血管事件 非系统性红斑狼疮患者的动脉粥样硬化。目的5)进行体外诱变和 表达不同载脂蛋白H等位基因异构体的研究 低密度脂蛋白氧化的异构体特异性抑制。

DESCRIPTION (provided by applicant): The risk of coronary heart disease (CHD) in systemic lupus erythematosus (SLE) women is up to 50 times higher than in the general population. The conventional risk factors are insufficient to explain premature CHD in SLE patients. Compared to about 1-5 percent prevalence of antiphospholipid antibodies (APA) in the general U.S. white population, about 50 percent of the SLE patients are positive for APA. ApoH is a principal autoantigen for the production of APA in patients with autoimmune diseases. ApoH inhibits the in vitro uptake of oxidized low-density lipoprotein (oxLDL) by macrophages, but in the presence of APA it promotes the ihflux of oxLDL into macrophages. As the accumulation of oxLDL in macrophages is believed to initiate the atherosclerotic process, these findings suggest that apoH-mediated immune response in patients with autoimmune diseases, like SLE, may lead to atherosclerosis. In this renewal we propose to examine the joint roles of APA, antibodies to oxLDL (anti-oxLDL) and APOH genetic variation (known and discovered as part of this proposal) in relation to the occurrence of CHD in SUE and non-SLE patients. Our hypothesis is that individuals positive for APA and/or anti-oxLDL are prone to premature CHD and this susceptibility is modified by common genetic variation in the APOH gene. The objectives of the study will be achieved by fulfilling the five aims. Aim 1) identify and characterize naturally occurring common mutations in all exons, introns and the 3' region of the APOH gene by polymerase chain reaction (PCR), denaturing HPLC analysis and DNA sequencing in SLE and non-SLE CHD patients, and African blacks positive for APA. Aim 2) determine the prevalence and correlation between APA (anti-apoH, anticardiolipin, lupus anticoagulant) and anti-oxLDL in plasma samples from SLE patients and controls. Aim 3) determine the relationship between APOH genetic variation (data generated in Aim 1) and the occurrence of APA and anti-oxLDL (data generated in Aim 2). Aim 4) examine the relationship between APOH genetic variation (data generated in Aim 1) and the occurrence of subclinical cardiovascular events in SLE patients and with coronary atherosclerosis in non-SLE patients. Aim 5) perform in vitro mutagenesis and expression studies to express different apoH allelic-isoforms to evaluate isoform-specific inhibition of LDL oxidation.