MHC Array T Cell Assay System for Monitoring Immune Status in Type 1 Diabetes

描述(由申请人提供)： 在美国，1型糖尿病(T1D)影响着100多万人，通常在儿童时期被诊断出来，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建议的目标是开发一种自动T细胞分析试验，以识别和功能表征自身反应性的T1D抗原特异的CD4和CD8 T细胞，能够使用仅2毫升的患者样本同时检测数千个靶点。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在一个独立的样本盒中结合MHC多肽阵列和一个高度并行的发光检测系统，该系统能够使用比目前方法小得多的样本体积对T1D抗原特异性T细胞进行快速、多路分析。该系统的主要进步将是能够直观地检测与MHC-多肽复合体结合的抗原特异性的CD4和CD8自身反应T细胞，并将MHC：T细胞受体结合与由此产生的多种细胞因子效应分子的激活和分泌联系起来。我们的目标是创建一种T细胞分析系统，该系统提供高通量筛查、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 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 autoreactive 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 autoreactive 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 proposal is to develop an automated T cell analysis assay to identify and functionally characterize autoreactive T1D antigen-specific CD4+ and CD8+ T cells capable of testing 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 of this project will combine MHC-peptide arrays in a self-contained sample cartridge with a highly parallel luminescent detection system that is capable of rapid, multiplexed analysis of T1D antigen-specific T cells using dramatically smaller sample volumes than current methods. The major advancements of this system will be the ability to visually detect antigen-specific CD4+ and CD8+ autoreactive T cells binding to MHC-peptide-complexes and to correlate MHC: T cell receptor binding to resulting activation and secretion of multiple cytokine effector molecules. The goal is to create a T cell analysis system that offers high throughput screening, multi-parametric 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.