Functional phenotyping of leukocyte reprogramming in Type 1 diabetes

描述(申请人提供)：1型糖尿病(T1D)在美国影响着100多万人{来源：ADA http://www.diabetes.org/about-diabetes.jsp}，通常在儿童中被诊断出来，可能导致失明、心脏病和肾衰竭。对T1D的干预的一个主要焦点是检测和鉴定自身反应性T细胞，它在攻击产生胰岛素的胰岛细胞中发挥核心作用。为了使对T1D的医疗干预最有效，应该在症状出现之前发现和治疗这种疾病。目前识别自身反应性T1D特异性T细胞靶点的方法速度慢、技术要求高、劳力和试剂密集，并且要消耗大量的T细胞来测试有限数量的靶点。样本量是T1D中一个极其重要的考虑因素，在检测临床样本时往往是一个限制因素，特别是在儿童和青少年无法提供大样本的情况下。这项SBIR应用的目标是开发一种自动T细胞分析测试，以识别和功能表征自动反应的T1D抗原特异性的CD4+和CD8+T细胞，能够使用仅2毫升的患者样本同时检测成百上千个目标。建议的系统是基于将正在申请专利的MHC-肽阵列技术与自动流动细胞检测和分析系统相结合。该项目的创新方法将结合一个独立的样品盒，其中抗原特异性的CD4+和CD8+自动反应T细胞与高亲和力的MHC多肽单体结合，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对已识别细胞亚群分泌的细胞因子的检测相结合。我们的目标是创建一种T细胞分析系统，该系统提供高含量筛选、多参数表征能力，并采用最先进的集成样本处理，以便于使用。目前，还没有可用的系统提供广泛的功能，使用的简单性和有限的样本量要求，如建议的T细胞分析系统使用MHC-肽阵列来对抗T1D。

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.