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

用于监测 1 型糖尿病免疫状态的 MHC 阵列 T 细胞检测系统

• 批准号: 7211919
• 负责人: Ernest Fitch Guignon
• 金额: $ 50.41万
• 依托单位: CIENCIA, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7211919
• 关键词: Address; Adolescent; Affect; Alleles; Amputation; Antibodies; Antigen-Presenting Cells; Antigens; Antihypertensive Agents; Arts; Autoantigens; Autoimmune Process; Autoimmunity; Benchmarking; Beta Cell; Binding; Bioinformatics; Biological Assay; Blindness; Blood specimen; Businesses; CD4 Antigens; CD8B1 gene; Cardiovascular Diseases; Cells; Child; Class; Clinical; Collaborations; Complex; Data; Detection; Development; Diabetes Mellitus; Diagnosis; Diagnostic; Disease; Elements; Encephalopathies; Endocrine; Epitopes; Fluorescence; Frequencies; Genes; Goals; Haplotypes; Heart Diseases; Histocompatibility Antigens Class II; Image; Immune; Immune system; Immunologic Monitoring; Immunotherapeutic agent; Incidence; Individual; Inflammatory; Inflammatory Response; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention; Islet Cell; Islets of Langerhans; Kidney Failure; Knowledge; Laboratories; Laboratory Procedures; Laboratory Research; Lasers; Lead; Legal patent; Libraries; Lipids; Major Histocompatibility Complex; Manuals; Marketing; Massachusetts; Mediating; Medical; Medical center; Metabolic; Methods; Monitor; Nerve; Noise; Numbers; Operative Surgical Procedures; Pain; Pathogenesis; Patients; Peptide Library; Peptides; Performance; Persons; Phase; Play; Population; Process; Production; Proteins; Reagent; Research; Research Institute; Risk; Role; Sample Size; Sampling; Services; Signal Transduction; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Specificity; Staging; Symptoms; Synthetic Antigens; System; Systems Analysis; T-Cell Immunologic Specificity; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; Techniques; Technology; Testing; Therapeutic; Time; United States; Universities; Virginia; Work; autoreactive T cell; base; charge coupled device camera; cognitive function; commercialization; cost; cytokine; design; diabetic; fight against; fluorescence microscope; high throughput screening; image visualization; immunoregulation; innovation; instrument; instrumentation; luminescence; medical schools; multiplex detection; novel; novel therapeutics; prototype; receptor binding; research and development; response; vaccination strategy

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.

描述（由申请人提供）： 1型糖尿病（T1 D）在美国影响超过100万人，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提案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联合收割机MHC-肽阵列与高度平行的发光检测系统相结合，该系统能够使用比当前方法小得多的样品体积快速、多路复用地分析T1 D抗原特异性T细胞。该系统的主要进步将是视觉检测抗原特异性CD 4+和CD 8+自身反应性T细胞与MHC-肽复合物结合的能力，以及将MHC：T细胞受体结合与多种细胞因子效应分子的活化和分泌相关联的能力。我们的目标是创建一个T细胞分析系统，提供高通量筛选，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。