An acoustofluidic avidity cytometer for massive parallel profiling single autoreactive T cell in autoimmune disease

项目摘要 自身反应性T细胞应答靶向中枢神经系统来源的自身抗原，并引起几种 脱髓鞘自身免疫性疾病，包括多发性硬化症、急性弥漫性脑脊髓炎和 视神经肌萎缩多发性硬化症是这些疾病中最常见的一种， 在全世界范围内并且在美国约有400，000人，是一种慢性炎性疾病，引起 脑和脊髓中的脱髓鞘病变和斑块。然而，目前的临床试验都不能 由于缺乏足够的诊断， 敏感和有效的生物标志物。 多发性硬化患者的T细胞显示出对髓鞘蛋白的亲和力增加的活化表型。 因此，我们假设自身反应性T细胞的亲合力是一种疾病标志物，可用于监测 疾病进展，判断治疗反应，或发现新的生化疾病标志物， 硬化症然而，目前的技术都没有实现高灵敏度和高通量 在单细胞水平上测量临床样品中的细胞亲合力。PI的工程进展， “声流体学”允许以意想不到的分辨率精确操纵单细胞和液体， 有前途的潜力，用于测量高度异质性，临床自身反应性T细胞的亲合力，以及 当与PI的“高通量”结合时， 微流控技术。 目的是建立一种声流亲合力流式细胞仪，以克服临床检测的障碍， 自身免疫性疾病如多发性硬化症。然而，有几个重大障碍，其中包括 建立一个高灵敏度和高通量的亲合力细胞仪来分析单个自身反应性髓鞘特异性T细胞 细胞并确定它们在临床样品中的亲合力分布。高通量微流体技术的发展将 允许针对新的自身抗原筛选感兴趣的单个自身反应性T细胞。拟议的仪器 将有可能彻底改变目前的临床测试和发现新的诊断，预后， 自身免疫性疾病包括多发性硬化症、类风湿性关节炎、克罗恩病等的治疗功效。

Project Summary Autoreactive T cell responses target central nervous system-derived autoantigens and cause several demyelinating autoimmune diseases including multiple sclerosis, acute disseminated encephalomyelitis, and neuromyelitis optica. Multiple sclerosis, the most common of these diseases affecting over two million people worldwide and approximately 400,000 people in United States, is a chronic inflammatory disease, causing lesions and plaques of demyelination in the brain and spinal cord. However, none of the current clinical tests can confidently predict clinical multiple sclerosis progression or treatment efficacy due to a lack of sufficiently sensitive and effective biomarkers. T cells in multiple sclerosis patients display an activated phenotype with an increased avidity to myelin protein. As a result, we hypothesize that the avidity of autoreactive T cells is a disease marker that can be used to monitor disease progress, judge therapeutic response, or discover new biochemical disease markers for multiple sclerosis. However, none of the current technologies have achieved high-sensitivity and high-throughput measurement of cell avidity, in clinical samples, at the single cell level. The PI’s engineering advances in ‘Acoustofluidics,’ allowing for precise manipulation of single cells and liquids at unexpected resolution, shows promising potential for measuring the avidity of highly heterogeneous, clinical autoreactive T cells as well as for potentially sorting and analyzing single cell of interest, when combined with the PI’s ‘High Throughput Microfluidic’ technology. The objective is to create an acoustofluidic avidity cytometer to overcome the barriers in detecting clinical autoimmune disorders such as multiple sclerosis. However, there are several significant hurdles which include building a high-sensitivity and high-throughput avidity cytometer to profile single autoreactive myelin-specific T cells and determine their avidity distribution in clinical samples. Incorporating high-throughput microfluidics will allow the screening of single autoreactive T cells of interest for new autoantigens. The proposed instrumentation will potentially revolutionize the current clinical testing and discovery new markers for diagnosis, prognosis, and treatment efficacy of autoimmune disease including multiple sclerosis, rheumatoid arthritis, Crohn’s disease, etc.