Functional phenotyping of leukocyte reprogramming in Type 1 diabetes

描述（由申请人提供）：1型糖尿病（T1 D）影响美国超过100万人{来源ADA http：//www.diabetes.org/about-diabetes.jsp}，通常在儿童中诊断，可导致失明、心脏病和肾衰竭。T1 D干预的主要焦点是自身反应性T细胞的检测和表征，其在攻击产生胰岛素的胰岛细胞中起核心作用。为了使T1 D的医疗干预最有效，应该在症状发作之前检测和治疗疾病。目前用于鉴定自身反应性T1 D特异性T细胞的靶标的方法是缓慢的，技术要求高，劳动和试剂密集型的，并且消耗大量T细胞来测试有限数量的靶标。样本量是T1 D的一个非常重要的考虑因素，在测试临床样本时通常是一个限制因素，特别是当幼儿和青少年无法提供大样本时。该SBIR应用的目的是开发一种自动化T细胞分析测定，以鉴定和功能表征自身反应性T1 D抗原特异性CD 4+和CD 8 + T细胞，该T细胞能够使用少至2 ml的患者样本同时检测数百至数千个靶标。所提出的系统是基于集成专利申请中的MHC-肽阵列技术与自动化流动池检测和分析系统。该项目的创新方法将结合联合收割机一个独立的样品盒，其中抗原特异性CD 4+和CD 8+自身反应性T细胞与高亲合力MHC-肽单体结合，并通过表面等离子体共振与相关的高灵敏度表面等离子体共振增强检测由鉴定的细胞亚群分泌的细胞因子进行检测。我们的目标是创建一个T细胞分析系统，提供高内容筛选，多参数表征能力，并结合了最先进的集成样品处理，易于使用。目前，还没有可用的系统提供广泛的能力，使用的简单性和有限的样本量要求，作为提出的T细胞分析系统使用MHC-肽阵列在对抗T1 D。

DESCRIPTION (provided by applicant): Type 1 Diabetes (T1D) affects more than 1 million people in the United States {Source ADA http://www.diabetes.org/about-diabetes.jsp} and is usually diagnosed in children and can lead to blindness, heart disease and kidney failure. A major focus of intervention for T1D is on the detection and characterization of auto-reactive T cells, which play a central role in the attack on insulin producing islet cells. For medical intervention for T1D to be most effective, the disease should be detected and treated before the onset of symptoms. Current methods to identify the targets of auto-reactive T1D-specific T cells are slow, technically demanding, labor- and reagent-intensive, and consume large numbers of T cells to test limited numbers of targets. Sample size is an extremely important consideration in T1D and is often a limiting factor when testing clinical samples, especially as young children and adolescents are unable to give large samples. The objective of this SBIR application is to develop an automated T cell analysis assay to identify and functionally characterize auto-reactive T1D antigen-specific CD4+ and CD8+ T cells capable of testing hundreds to thousands of targets simultaneously using as little as 2ml of patient sample. The proposed system is based upon integrating patent pending MHC-peptide array technology with an automated flow cell detection and analysis system. The innovative approach to this project will combine a self-contained sample cartridge in which antigen-specific CD4+ and CD8+ auto-reactive T cells bind to high avidity MHC-peptide monomers and are detected via surface plasmon resonance with correlated highly sensitive surface plasmon resonance-enhanced detection of cytokines secreted by identified cell subpopulations. The goal is to create a T cell analysis system that offers high content screening, multi-parameter characterization capability and incorporates state-of- the-art integrated sample handling for ease-of-use. At present, there are no systems available that offer the breadth of capabilities, the simplicity of use and limited sample size requirements as the proposed T cell analysis system using MHC-peptide arrays in the fight against T1D.