Exosome-induced innate and antigen-specific immune responses in Type 1 Diabetes

1 型糖尿病中外泌体诱导的先天性和抗原特异性免疫反应

• 批准号: 8837261
• 负责人: YANG D. DAI
• 金额: $ 14.36万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-02 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8837261
• 关键词: Exosome; induced; innate; antigen; specific

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) is a T cell-mediated, tissue-specific autoimmune disease, in which the class II MHC genes play a major role controlling the disease susceptibility. This suggests that islet autoantigens and their short peptides presented on the MHC molecules are the key elements leading to the destruction of the insulin-producing beta cells in the pancreatic islets. Since self-tolerance mechanism in the thymus normally is effective in removing strong autoreactive T cells, peripheral circulating T cells are purged to avoid self- reactivity; however, autoimmune disease does occur despite of rare incidence. We hypothesized that certain inflammatory triggers may initiate the autoimmune responses in the islets by enhancing the immunogenic activity of the islet autoantigens. We have studied one candidate islet antigen, glutamic acid decarboxylase 65 KDa (GAD65), for several years and learned that unique antigen delivery and processing pathways are required to induce disease-causing or diabetogenic effector T cells, and demonstrated that a diabetes- associated candidate gene, Slc11a1, could alter the pathways of processing islet antigens in dendritic cells (DC). We recently isolated one type of microparticles/vesicles, namely exosomes (EXO), from the culture supernatants of mouse insulinomas and primary islet glial cells, and found that the EXO contains strong proinflammatory materials that could stimulate various antigen presenting cells including DC to produce inflammatory cytokines and upregulate MHC and costimulatory molecules; more importantly, the EXO collected from the primary islet glial cells also expresses several known islet autoantigens including GAD65. We thus propose a novel hypothesis that EXO may act as a unique type of endogenous adjuvant and antigen carrier that can condition APC to alter the processing/presenting of the autoantigens and thus, to activate the autoreactive T cells. In this proposal, we will first perform basic biochemistry analysis for the EXO released by the primary islet cells, and study the requirements of different TLRs in the EXO-induced innate responses to characterize the nature of the proinflammatory materials enclosed in the EXO. We will compare the islet glial cells and their EXO between diabetes-susceptible NOD strain and resistant strains to address whether abnormal or excess EXO production could be one diabetes-causative event. Then, we will use EXO as an antigen carrier to study how autoreactive effector T cells specific for islet antigens, particularly GAD65, are induced following processing the EXO in DC. We predict that increased leakage of islet antigens via EXO secretion under stress or inflammatory situation is the key for activating highly diabetogenic T cells. Finally, we will continue studying the Slc11a1 gene for its roles in DC activation and antigen processing. We will use EXO as a vehicle for activating DC and delivering antigens. We anticipate that EXO can upregulate Slc11a1 function in DC and thus promote induction of the disease-causing Th1 cells.

描述（由申请人提供）：1型糖尿病（T1 D）是一种T细胞介导的组织特异性自身免疫性疾病，其中II类MHC基因在控制疾病易感性方面发挥主要作用。这表明，胰岛自身抗原及其在MHC分子上呈递的短肽是导致胰岛中产生胰岛素的β细胞破坏的关键因素。由于胸腺中的自身耐受机制通常有效去除强自身反应性T细胞，因此清除外周循环T细胞以避免自身反应性;然而，尽管发生率很低，但自身免疫性疾病确实发生。我们假设某些炎症触发因子可能通过增强胰岛自身抗原的免疫原性活性来启动胰岛中的自身免疫应答。我们已经研究了一种候选胰岛抗原，谷氨酸脱羧酶65 kDa（GAD 65），多年来，并了解到独特的抗原递送和加工途径需要诱导致病或致糖尿病效应T细胞，并证明糖尿病相关的候选基因Slc 11 a1，可以改变树突状细胞（DC）中加工胰岛抗原的途径。我们最近从小鼠胰岛素瘤和原代胰岛胶质细胞的培养上清中分离到一种微粒/囊泡，即exosomes（EXO），发现EXO含有强烈的促炎物质，能刺激包括DC在内的多种抗原呈递细胞产生炎性细胞因子，上调MHC和共刺激分子;更重要的是，从原代胰岛神经胶质细胞收集的EXO也表达几种已知的胰岛自身抗原，包括GAD 65。因此，我们提出了一个新的假设，EXO可能作为一种独特类型的内源性佐剂和抗原载体，可以条件APC改变自身抗原的加工/呈递，从而激活自身反应性T细胞。在这个提议中，我们将首先对原代胰岛细胞释放的EXO进行基本的生物化学分析，并研究EXO诱导的先天性反应中不同TLR的需求，以表征EXO中封闭的促炎物质的性质。我们将比较糖尿病易感的NOD菌株和耐药菌株之间的胰岛胶质细胞和它们的EXO，以解决异常或过量的EXO产生是否可能是糖尿病的一个致病事件。然后，我们将使用EXO作为抗原载体来研究在DC中处理EXO后如何诱导对胰岛抗原特异性的自身反应性效应T细胞，特别是GAD 65。我们预测，在应激或炎症情况下，通过EXO分泌增加的胰岛抗原泄漏是激活高度致糖尿病性T细胞的关键。最后，我们将继续研究Slc 11 a1基因在DC活化和抗原加工中的作用。我们将使用EXO作为活化DC和递送抗原的载体。我们预期EXO可以上调DC中Slc 11 a1的功能，从而促进致病性Th 1细胞的诱导。