Effect of early diet and virus infections on immune regulation and the development islet autoimmunity -Trial to Reduce IDDM in the Genetically at Risk (TRIGR)

早期饮食和病毒感染对免疫调节和胰岛自身免疫发育的影响 - 减少遗传风险 (TRIGR) 中 IDDM 的试验

基本信息

批准号：
8970936
负责人：
Suvi Mirjami Virtanen
金额：
$ 109.58万
依托单位：
NATIONAL INSTITUTE FOR HEALTH & WELFARE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-10 至 2019-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Effect of early diet and virus infections on immune regulation and the development islet autoimmunity -Trial to Reduce IDDM in the Genetically at Risk (TRIGR). The study will yield important new information about the main environmental candidate risk factors in the pathogenesis of type 1 diabetes (T1D) utilizing the setup of TRIGR (dietary intervention). The aim is to evaluate associations between n-3 fatty acids, vitamin D, cow's milk exposure and viral infections with indicators of immune regulation and inflammation as well as induction of islet autoimmunity. Islet autoimmunity is defined as repeated positivity fo at least two diabetes-associated autoantibodies out of four ones measured. The current study will be based on biosamples collected in the TRIGR cohort. TRIGR study is an international double-blind randomized clinical trial of 2159 infants with HLA-conferred disease susceptibility and a first-degree relative with T1D recruited between 2002-2007 in 15 countries. In TRIGR an extensively hydrolyzed casein formula is compared to regular cow's milk based one. All subjects are followed until the youngest child will be 10-year-old. Blood samples are collected at 3 to 12 months' intervals. We measure from serum in cohort or nested case-control design: fatty acid composition with gas chromatography; 25-OH-vitamin D concentration with chemiluminescent mircoparticle immunoassay; virus antibodies with plaque neutralization and enzyme immune assay; cow's milk antibodies with enzyme-linked immunosorbent assay; antibodies to dietary bovine insulin by using the competitive radio-immunoassay with radioactive and cold insulin; and cytokines and chemokines with the Milliplex MAP kit. When studying cell-mediated responses to Coxsackievirus B, ß-lactoglobulin and bovine insulin, we use frozen peripheral blood mononuclear cells (PBMC), from which we measure Treg Th1, Th2 and Th17 markers, FoxP3, CTLA-4, IFN-g, IL-4, IL-5, IL-13, IL-17 and IL-22, using RT-qPCR methodology in cow's milk protein stimulated PBMCs. We will also analyze from frozen PBMCs the epigenetic modulation of two genes (FOXP3, IL-2) responsible for the development of regulatory T-cells. The methods are very well established and well-functioning in our laboratories. The analysis can start straight after the samples are received from TRIGR. This application covers the costs for the analysis of serum 25-OH-vitamin D concentration, serum fatty acids, immune regulation, inflammatory markers, and virus infections. It also covers coordination, data collection, management and statistical analysis as well as sample collection and aliquation for the current study.

描述（由申请人提供）：早期饮食和病毒感染对免疫调节和胰岛自身免疫发育的影响 - 减少遗传风险中 IDDM 的试验（TRIGR）该研究将产生有关主要环境候选风险因素的重要新信息。利用 TRIGR（饮食干预）的设置来研究 1 型糖尿病 (T1D) 的发病机制，目的是通过指标评估 n-3 脂肪酸、维生素 D、牛奶暴露与病毒感染之间的关联。胰岛自身免疫被定义为测量的四种糖尿病相关自身抗体中至少有两种呈阳性。当前的研究将基于 TRIGR 研究中收集的生物样本。是一项国际双盲随机临床试验，招募了 2159 名具有 HLA 赋予疾病易感性的婴儿以及患有 T1D 的一级亲属2002 年至 2007 年，TRIGR 对水解酪蛋白配方奶粉与普通牛奶配方奶粉进行了比较，直到最小的孩子 10 岁为止，每隔 3 至 12 个月采集一次血液样本。我们通过队列或巢式病例对照设计测量血清：使用气相色谱法测定脂肪酸组成；使用化学发光微粒测定 25-OH-维生素 D 浓度；免疫测定；用斑块中和和酶免疫测定法测定牛奶抗体；用放射性和冷胰岛素以及细胞因子和趋化因子进行竞争性放射免疫测定法测定膳食牛胰岛素抗体；在研究细胞介导的对柯萨奇病毒 B、β-乳球蛋白和牛胰岛素的反应时，我们使用冷冻外周血单核细胞 (PBMC)，从中测量 Treg我们使用 RT-qPCR 方法在牛奶蛋白刺激的 PBMC 中检测 Th1、Th2 和 Th17 标记物、FoxP3、CTLA-4、IFN-g、IL-4、IL-5、IL-13、IL-17 和 IL-22。还将分析冷冻 PBMC 中负责调节性 T 细胞发育的两个基因（FOXP3、IL-2）的表观遗传调节。这些方法在我们的研究中已经非常成熟且运作良好。从 TRIGR 收到样品后即可立即开始分析。该应用程序涵盖了血清 25-OH-维生素 D 浓度、血清脂肪酸、免疫调节、炎症标志物和病毒感染的分析费用。当前研究的协调、数据收集、管理和统计分析以及样本收集和分配。