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

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(申请人提供):早期饮食和病毒感染对免疫调节的影响和胰岛自身免疫的发展-减少遗传风险中的IDDM的试验(TRIGR)。这项研究将利用TRIGR(饮食干预)的设置,提供关于1型糖尿病(T1D)发病机制中主要环境候选危险因素的重要新信息。其目的是通过免疫调节和炎症指标以及胰岛自身免疫诱导来评估n-3脂肪酸、维生素D、牛奶暴露与病毒感染之间的关系。胰岛自身免疫的定义是在测量的四种糖尿病相关自身抗体中,至少有两种重复阳性。目前的研究将基于在TRIGR队列中收集的生物样本。TRIGR研究是一项国际双盲随机临床试验,在15个国家和地区招募了2159名患有人类白细胞抗原相关疾病易感性的婴儿,并且是T1D的一级亲属。在TRIGR中,广泛水解的酪蛋白配方与普通牛奶配方进行了比较。所有受试者都被跟踪,直到最小的孩子10岁。每隔3至12个月采集一次血样。我们采用队列或嵌套病例对照设计对血清进行检测:用气相色谱测定脂肪酸组成;用化学发光镜片免疫分析法测定25-OH-维生素D浓度;用斑块中和法和酶免疫法测定病毒抗体;用酶联免疫吸附分析法测定牛奶抗体;用放射免疫法和冷胰岛素竞争性放射免疫法测定饲料中抗牛胰岛素的抗体;用Milliplex MAP试剂盒测定细胞因子和趋化因子。在研究细胞对柯萨奇病毒B、乳球蛋白和牛胰岛素的反应时,我们使用冰冻的外周血单个核细胞(PBMC),用RT-qPCR方法检测牛乳蛋白刺激的PBMC中的Treg Th1、Th2和Th17标志、FoxP3、CTLA-4、IFN-g、IL-4、IL-5、IL-13、IL-17和IL-22。我们还将分析冷冻的PBMC中负责调节性T细胞发育的两个基因(FOXP3和IL-2)的表观遗传调节。这些方法在我们的实验室中得到了很好的确立和运作。分析可以在从TRIGR收到样品后立即开始。这项应用涵盖了分析血清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成本。它还包括本研究的协调、数据收集、管理和统计分析以及样本收集和整理。

项目成果

期刊论文数量(3)
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