Core B: Immunoassay Core

核心B项目摘要 核心B将与四个项目中的每一个合作，在小鼠和人类中提供相关的免疫测定： 项目1与赫德里克博士将使用小鼠模型来测试新的单核细胞亚群的影响， 调节动脉壁中的T和B细胞反应、动脉粥样硬化形成和先天免疫。核心将能够 测量氧化特异性表位（OSE）和ApoB-免疫复合物的IgG和IgM自身抗体， 以及总IgG和IgM血浆水平，以评估这些单核细胞亚群的影响以及与 淋巴细胞在这些测量。然后可以根据B1和B2细胞功能分析这些细胞， 博士McNamara的项目，也与Ley博士的实验中单核细胞细胞因子的释放相关。 米勒博士的项目2将使用核心来测量个体 脂蛋白，一种由我们实验室与米勒博士共同开发的测定方法， 临床数据在其章节中提供。我们已经建立的临床检测，如OxPL-apoB，Lp（a）， 还将测量IgG和IgM自身抗体以及ApoB-免疫复合物。这些措施可以 然后与冠状动脉钙（CAC）的存在和程度以及 CAC及其与心血管事件的关系。此外，它们可以与AIBP水平相关， 来自各种实验的HDL参数。 麦克纳马拉博士的项目3将评估调节它们生产能力的机制， 动脉粥样硬化保护部分如IgM至OSE。该项目预计将在B1中产生重要数据 在小鼠模型中，所有这些都与测量IgG和IgM自身抗体滴度高度相关。我们 也有能力测量这种小鼠中动脉粥样硬化保护抗体E06的存在， 获得针对E06、AB 1 -2的抗独特型抗体。最后，测量IgG和IgM抗体的总水平是 这是解释实验研究的一个重要方面，也将被衡量。此类测定法是 目前可用，并且先前已在与McNamara博士的合作中描述和发表 Ley博士的项目4将确定小鼠中ApoB特异性CD 4 T细胞的大小和性质。的 小鼠apoB-免疫复合物测定使用鼠单克隆抗体LF 3捕获鼠apoB 然后评估是否存在与apoB结合的抗体。这些分析将与此高度相关 在解释是否含有鼠apoB的免疫复合物在这些小鼠中产生的工作。 此外，我们可以用这些测定法测量鼠总apoB水平作为对照测量。

Core B Project Summary Core B will work with each of the four projects to provide relevant immunoassays in mice and in humans: Project 1 with Dr Hedrick will use mouse models to test the impact of new monocyte subsets that regulate T and B cell responses in the artery wall, on atherogenesis and innate immunity. The Core will be able to measure IgG and IgM autoantibodies to oxidation-specific epitopes (OSE) and ApoB-immune complexes, as well as total IgG and IgM plasma levels to assess the impact of these monocyte subsets and interaction with lymphocytes on these measurements. These can then be analyzed according to B1 and B2 cell function from Dr. McNamara's projects and to also correlate with monocyte cytokine release from Dr. Ley's experiments. Project 2 with Dr. Miller will use the Core to measure oxidized cholesteryl esters on individual lipoproteins, an assay that has been co-developed by our laboratory with Dr. Miller and for which preliminary clinical data is provided in his section. Clinical assays that we have established, such as OxPL-apoB, Lp(a), and IgG and IgM autoantibodies and ApoB-immune complexes will also be measured. These measures can then be related to both the presence and extent of coronary artery calcium (CAC), as well as the progression of CAC and the relationship with cardiovascular events. Furthermore, they can be correlated with AIBP levels and HDL parameters from the various experiments. Project 3 with Dr McNamara will evaluate the mechanism that regulates their ability to produce atheroprotective moieties such as IgM to OSE. This project is expected to generate significant data in B1 biology in mouse models, all of which will be highly relevant to measuring IgG and IgM autoantibody titers. We also have the ability to measure the presence of the atheroprotective antibody E06 in such mice as we have access to an anti-idiotype antibody to E06, AB1-2. Finally, measuring total levels of IgG and IgM antibodies is an important aspect in interpreting experimental studies, which will also be measured. Such assays are currently available and have been previously described and published in collaborative work with Dr. McNamara Project 4 with Dr. Ley will define the size and nature of the ApoB-specific CD4 T cell in mice. The mouse apoB-immune complex assays capture murine apoB using murine monoclonal antibody antibody LF3 and then assess the presence of a bound antibody to the apoB. These assays will be highly relevant to this work in interpreting whether immune complexes containing murine apoB are generated in these mice. Furthermore, we can measure murine total apoB levels with these assays as a control measure.