Multiple Autoantigens, Multiple Epitopes of Type 1 Diabetes

项目摘要 我们研究的首要目标是开发新的胰岛自身免疫生物标志物，并将其翻译成 预防人类1型糖尿病（T1D）的新发现。在上一个融资期间，我们开发了一个 检测胰岛和非胰岛自身抗原的血清自身抗体的多重测定，具有优异的灵敏度 和预测价值相比，我们的传统的放射性结合分析。拟议的研究将扩大 相关自身抗原的谱与那些后修饰的（PTM）以及提供 深入了解T1D的自然史中天然与PTM结合自身抗体何时发展。我们最近 这些发现加强了自身反应性T细胞对（原）胰岛素反应的证据，包括那些 由杂合胰岛素肽（HIP）和缺陷核糖体胰岛素产物（DRiPs）激活，在剩余的 T1D器官供体的胰岛。然而，我们仍然缺乏强有力的T细胞生物标志物，可以反映 自身反应性T细胞的活性，特别是在胰岛自身免疫的最早阶段或在免疫调节期间 以防止进展为临床糖尿病。检测胰岛自身抗体和自身反应性T细胞， 天然的和修饰的胰岛自身抗原，结合对它们之间关系的更好理解， 提高我们对T1D发病机制的认识，并改善对T1D各阶段进展的预测。 在具体目标1中，我们将优化和多重自身抗体测定天然和PTM修饰的胰岛 自身抗原，以确定这些抗体在儿童纵向样本中的时间发展 从出生到胰岛自身免疫和临床糖尿病的发展进行前瞻性随访。具体目标2 专注于开发从全血DNA测量的胰岛自身免疫的非细胞T细胞生物标志物 使用从T1D器官供体的残余胰岛获得的T细胞受体序列。成功 该提案的完成将导致用于测量胰岛自身抗体的可扩展的测定， 自身反应性T细胞，并提高对自身抗体和T细胞免疫的时间的理解 在T1D的发展过程中对天然和免疫后修饰的胰岛自身抗原的反应。

Project Summary The overarching goal of our research is to develop new biomarkers of islet autoimmunity and to translate these discoveries to the prevention of human type 1 diabetes (T1D). During the last funding period, we developed a multiplexed assay to detect serum autoantibodies to islet and non-islet autoantigens with excellent sensitivity and predictive value when compared to our traditional radio-binding assays. The proposed studies will broaden the spectrum of relevant autoantigens to those that are post-translationally modified (PTM) as well as provide insights into when native versus PTM binding autoantibodies develop in the natural history of T1D. Our recent findings have strengthened the evidence for autoreactive T cells responding to (pro)insulin, including those activated by hybrid insulin peptides (HIPs) and defective ribosomal insulin products (DRiPs), within the residual pancreatic islets of T1D organ donors. However, we still lack robust T cell biomarkers that could reflect the activity of autoreactive T cells, especially at the earliest stages of islet autoimmunity or during immunomodulation to prevent progression to clinical diabetes. Assays for islet autoantibodies and autoreactive T cells that measure both natural and modified islet autoantigens, combined with a better understanding of their relationship, will enhance our knowledge of T1D pathogenesis and improve prediction of progression through the stages of T1D. In specific aim 1, we will optimize and multiplex autoantibody assays to native and PTM modified islet autoantigens to determine the temporal development of these antibodies in longitudinal samples from children prospectively followed from birth to development of islet autoimmunity and clinical diabetes. Specific aim 2 focuses on developing a non-cellular T cell biomarker of islet autoimmunity measured from whole blood DNA using the T cell receptor sequences obtained from residual islets of T1D organ donors. The successful completion of this proposal will result in scalable assays for the measurement of islet autoantibodies and autoreactive T cells, and improved understanding regarding the timing of autoantibody and T cell immune responses to native and post-translationally modified islet autoantigens during the development of T1D.