Analysis of Food Specific T cells by a Novel Microengraving Technology

通过新型微雕刻技术分析食物特异性 T 细胞

• 批准号: 8039134
• 负责人: John Christopher Love
• 金额: $ 19.3万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8039134
• 关键词: Address; Allergens; Allergic; Antigens; Biological; Biological Assay; CD8B1 gene; Cell Count; Cell Separation; Cells; Characteristics; Child; Clinical; Clinical assessments; Collaborations; Complement; Devices; Diagnostic; Enzymes; Event; Evolution; Exposure to; Fluorescence-Activated Cell Sorting; Food; Food Analysis; Food Hypersensitivity; Frequencies; Genotype; Glycoproteins; Goals; Human Engineering; Hypersensitivity; Immune; Immunologic Monitoring; Immunology; Individual; Knowledge; Lead; Link; Longitudinal Studies; Love; Lymphocyte; Measures; Methodology; Methods; Milk; Monitor; Oral; Patients; Pediatric Hospitals; Prevalence; Process; Public Health; Reaction; Research; Role; Sampling; Sensitivity and Specificity; Stimulus; Surface; T-Lymphocyte; Techniques; Technology; Testing; To specify; Translating; Western World; analytical tool; base; cytokine; desensitization; food allergen; improved; innovation; insight; interest; new technology; novel; novel diagnostics; oral tolerance; peripheral blood; research study; response; skin prick test; therapy development; translational approach

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.

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，其患病率以非常显著的速度增加，特别是在儿童中。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开发一种使用新型微技术表征过敏原特异性T细胞的测定方法，该方法允许对单个淋巴细胞进行高通量，灵敏的分析。我们将应用这种新的检测方法来评估食物过敏个体中食物特异性T细胞的频率，然后监测接受口服牛奶脱敏的儿童中食物特异性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：检测单个克隆功能反应所需的灵敏度，以及处理儿童临床样本中少量细胞的能力。该项目是Love实验室（MIT）与Umetsu实验室（儿童医院）之间的合作，该实验室具有对来自>10^5个单细胞的分泌产物进行同时平行分析的专业知识，用于过敏患者的临床分析和免疫学评估。这个跨学科的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，这种翻译方法将提高我们对食物过敏和耐受性的免疫学基础以及免疫偏离、耐受性和过敏原特异性T细胞缺失的作用的认识。该项目的具体目标是：1）开发一种使用单细胞显微技术评估食物特异性T细胞的测定方法; 2）研究口服耐受性形成的特异性免疫机制。建立定量分析工具也可能导致新的临床诊断，补充挑战试验或皮肤点刺试验，并使其他干预性研究的有效性的常规免疫监测。食物过敏是公共卫生的一个越来越大的负担，特别是在我们的儿童中，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对牛奶快速脱敏的生物机制。这些结果对于评估这种疗法的实用性非常重要，并且本研究中使用的基础技术的发展也可能导致用于监测过敏治疗的新诊断。