Functional phenotyping of leukocyte reprogramming in Type 1 diabetes

1 型糖尿病白细胞重编程的功能表型

• 批准号: 8402800
• 负责人: Ernest Fitch Guignon
• 金额: $ 26.48万
• 依托单位: CIENCIA, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-18 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8402800
• 关键词: Address; Adolescent; Affect; Antigens; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Avidity; Beta Cell; Binding; Biological Assay; Biological Markers; Blindness; Blood specimen; Body part; CD8B1 gene; Cell Line; Cells; Child; Clinical; Coupled; Cultured Cells; Detection; Development; Diabetes Mellitus; Diagnosis; Disease; Disease Progression; Early Diagnosis; Engineering; Generations; Goals; Heart Diseases; Heat shock proteins; Human; Image; Immobilization; Immune system; Inbred NOD Mice; Individual; Inflammatory Response; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention; Islet Cell; Islets of Langerhans; Kidney Failure; Knowledge; Lead; Legal patent; Leukocytes; Measurement; Measures; Mediating; Medical; Metabolic syndrome; Methods; Mitogens; Modality; Modeling; Monitor; Mus; Nature; Onset of illness; Optics; Pancreas; Patients; Peptide/MHC Complex; Performance; Phase; Phenotype; Play; Population; Predictive Value; Protocols documentation; Reagent; Risk; Role; Sample Size; Sampling; Screening procedure; Sensitivity and Specificity; Serum; Signal Transduction; Small Business Innovation Research Grant; Source; Surface Plasmon Resonance; Symptoms; System; Systems Analysis; T-Lymphocyte; Technology; Testing; Therapeutic Intervention; Time; United States; Whole Blood; base; cytokine; design; diabetes mellitus therapy; experience; fight against; in vivo; innovation; instrument; instrumentation; monomer; mouse model; next generation; prototype; response; sensor; stress protein; surface plasmon coupled emission

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. PUBLIC HEALTH RELEVANCE: Type 1 Diabetes (T1D) is an autoimmune disease usually diagnosed in children where T cells, part of the body's own immune system, attack pancreas cells that make insulin. Detecting auto-reactive T cells with current methods is impractical, time consuming and requires large patient samples. This application is to develop a high throughput T cell analysis system to detect and characterize auto-reactive T1D T cells using substantially smaller samples. Knowledge gained by the use of this T cell analysis system may lead to the faster discovery of T1D T cell targets, the ability to better monitor T1D therapies in clinical trils and the potential for earlier diagnosis of T1D.

描述（由申请人提供）：1型糖尿病（T1D）在美国影响超过100万人{来源ADA http://www.diabetes.org/about-diabetes.jsp}，通常在儿童中被诊断出来，可导致失明、心脏病和肾衰竭。T1D干预的一个主要焦点是检测和表征自身反应性T细胞，它在攻击产生胰岛素的胰岛细胞中起着核心作用。为了使T1D的医疗干预最有效，疾病应该在症状出现之前被发现和治疗。目前用于识别自身反应性t1d特异性T细胞靶标的方法是缓慢的，技术要求高，劳动和试剂密集，并且消耗大量T细胞来测试有限数量的靶标。在T1D中，样本量是一个非常重要的考虑因素，并且在测试临床样本时往往是一个限制因素，特别是当年幼的儿童和青少年无法提供大样本时。此SBIR应用程序的目的是开发一种自动T细胞分析方法，以识别和功能表征自身反应性T1D抗原特异性CD4+和CD8+ T细胞，能够同时使用少至2ml的患者样本测试数百到数千个靶标。该系统基于正在申请专利的mhc肽阵列技术与自动流动池检测和分析系统的集成。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CD4+和CD8+自身反应性T细胞与高亲和度的mhc肽单体结合，并通过表面等离子体共振和相关的高度敏感的表面等离子体共振检测来检测由鉴定的细胞亚群分泌的细胞因子。目标是创建一个T细胞分析系统，提供高含量筛选，多参数表征能力，并结合最先进的集成样品处理，便于使用。目前，还没有一种系统可以提供广泛的功能，使用简单性和有限的样本量要求，如所提出的使用mhc肽阵列的T细胞分析系统，用于对抗T1D。