Analysis of Apolipoprotein H in Lupus

描述（由申请人提供）：冠心病（CHD）的风险 在系统性红斑狼疮（SLE）的妇女高达50倍以上， 普通民众。传统的风险因素不足以 解释SLE患者的早发冠心病相比之下， 抗磷脂抗体（阿帕）在一般美国白色人群中， 约50%的SLE患者阿帕阳性。ApoH是一个主要的 自身免疫性疾病患者体内阿帕产生的自身抗原。 ApoH抑制氧化低密度脂蛋白（oxLDL）的体外摄取 巨噬细胞，但在阿帕的存在下，它促进了oxLDL进入 巨噬细胞由于巨噬细胞中oxLDL的积累被认为是 启动动脉粥样硬化过程，这些发现表明apoH介导的 自身免疫性疾病（如SLE）患者的免疫反应可能导致 动脉粥样硬化在这次更新中，我们建议审查阿帕的联合作用， oxLDL抗体（抗oxLDL）和APOH遗传变异（已知和 作为本提案的一部分发现）与CHD的发生有关， SUE和非SLE患者。我们的假设是阿帕阳性的个体 和/或抗氧化低密度脂蛋白倾向于早发冠心病，这种易感性 由APOH基因中常见的遗传变异修饰。的目标 通过实现这五个目标来实现学习。（1）识别和 表征所有外显子、内含子和外显子中天然存在的常见突变， 通过聚合酶链反应（PCR）、变性HPLC测定APOH基因的3'区 SLE和非SLE CHD患者以及非洲黑人中的分析和DNA测序 阿帕阳性目的2）确定阿帕的患病率和与 （抗apoH、抗心磷脂、狼疮抗凝剂）和血浆中的抗oxLDL 来自SLE患者和对照的样品。（3）确定关系 APOH遗传变异（目标1中生成的数据）与 阿帕和抗oxLDL（目标2中生成的数据）。（4）检查关系 APOH遗传变异（目标1中生成的数据）与 SLE患者和冠心病患者的亚临床心血管事件 非SLE患者的动脉粥样硬化。目的5）进行体外诱变， 表达研究，以表达不同的apoH等位基因亚型， LDL氧化的亚型特异性抑制。

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.