Analysis of Food Specific T cells by a Novel Microengraving Technology

描述（由申请人提供）：食物过敏是西方世界的一个主要公共卫生问题，其患病率正以非常显著的速度增长，尤其是在儿童中。此外，我们对导致这些疾病的基本免疫机制的理解仍然很差，部分原因是我们研究这些问题的方法相对粗糙。本研究的总体目标是利用一种新的微技术开发一种表征过敏原特异性T细胞的检测方法，该技术允许对单个淋巴细胞进行高通量、敏感的分析。我们将应用这种新方法来评估食物过敏个体中食物特异性T细胞的频率，然后监测口服牛奶脱敏儿童中食物特异性T细胞功能多样性的演变。我们的技术解决了两个严重阻碍儿童过敏原特异性T细胞详细免疫学研究的挑战：检测单个克隆的功能反应所需的敏感性以及处理儿童临床样本中可用的少量细胞的能力。该项目是Love实验室（MIT）与Umetsu实验室（儿童医院）之间的合作，该实验室具有同时平行分析bbbb10 ^5单细胞分泌产物的专业知识，而Umetsu实验室（儿童医院）用于过敏患者的临床分析和免疫学评估。这个跨学科团队是独特和创新的，因为它结合了工艺工程和人类免疫学方面的大量专业知识，以解决理解和治疗食物过敏的挑战。总之，这种转化方法将提高我们对食物过敏和耐受性的免疫学基础的认识，以及免疫偏差、耐受性和过敏原特异性T细胞缺失的作用。该项目的具体目标是：1)开发一种利用单细胞微技术评估食物特异性T细胞的检测方法；2)研究口腔耐受产生的特定免疫机制。建立定量分析工具也可能导致新的临床诊断，以补充激发试验或皮肤点刺试验，并使常规免疫监测其他介入性研究的疗效成为可能。食物过敏对公众健康的负担越来越大，尤其是对我们的孩子，但是现有的脱敏治疗的机制还没有得到很好的理解。该项目将开发和应用一种基于微加工设备的新技术，以研究通过口服治疗控制牛奶快速脱敏的生物学机制。该结果对于评估该疗法的效用非常重要，并且本研究中使用的基础技术的发展也可能导致监测过敏治疗的新诊断方法。

DESCRIPTION (provided by applicant): Food allergy is a major public health problem in the Western world that is increasing in prevalence at a very significant rate, particularly among children. Moreover, our understanding of the basic immunological mechanisms that lead to these conditions remains poor, in part because our methodologies to study these problems are relatively crude. The overall goal of this research is to develop an assay for characterizing allergen-specific T cells using a novel microtechnology that allows high-throughput, sensitive profiling of individual lymphocytes. We will apply this new assay to evaluate the frequency of food-specific T cells in food- allergic individuals, and then to monitor the evolution of the functional diversity of food-specific T cells in children undergoing oral milk desensitization. Our technology addresses two challenges that significantly impede detailed immunological studies of allergen-specific T cells in children: the sensitivity required to detect functional responses from individual clones and the ability to handle the small numbers of cells that are available in clinical samples from children. This project is a collaboration between the Love lab (MIT) with expertise for simultaneous parallel analyses of secretory products from >10^5 single cells and the Umetsu lab (Children's Hospital) for clinical analysis and immunological assessments of allergy patients. This interdisciplinary team is unique and innovative in that it combines substantial expertise in both process engineering and human immunology to address the challenges of understanding and treating food allergies. Together, this translational approach will improve our knowledge of the immunological basis of food allergy and tolerance, and of the role of immune deviation, tolerance and deletion of allergen-specific T cells. The specific aims of this project are: 1) to develop an assay to evaluate food-specific T cells using a single-cell microtechnology and 2) to investigate the specific immunological mechanisms by which oral tolerance develops. Establishing quantitative analytical tools may also lead to new clinical diagnostics that complement challenge testing or skin-prick testing, and that enable routine immunological monitoring of the efficacy of other interventional studies. Food allergies are an increasing burden on public health, especially among our children, but the mechanisms by which existing treatments for desensitization operate are not well understood. This project will develop and apply a novel technology based on microfabricated devices to examine the biological mechanisms that govern rapid desensitization to milk by oral treatment. The results will be important for evaluating the utility of this therapy, and the development of the underlying technology used in this research may also lead to new diagnostics for monitoring allergy treatments.