HUMAN GENETIC ANALYSIS RESOURCE

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 基于阵列的抗原特异性T细胞的检测。 马克·戴维斯，P.I.，钱越秀，帕特里克·布朗和合作伙伴 概述。检测和定量人类外周血中抗原特异性T细胞的能力 四聚肽/MHC染色试剂已被证明对以下疾病非常有用 进展和评估干预的效果。然而，这项技术需要荧光 激活的细胞分选器，这严重限制了吞吐量，特异性的数量可以 调查和自动化的潜力。另外，它需要比较多的血液， 这对来自幼儿的样本来说是一个特别的问题(如前面的建议所指出的)。至 促进分析在感染流感病毒期间产生的免疫反应，并分析 疫苗接种的效果，我们已经产生了初步结果，表明四聚体可以排列。 在玻片上保留抗原特异性T细胞，保留的T细胞可以用荧光检测到 显微镜。该应用程序的目标是扩展和提炼这些结果以开发高吞吐量， 基于阵列的抗原特异性T细胞定量分析，将允许评估许多 同一血样具有不同的T细胞特性。这应该会提供一个方便的和潜在的 测量多个样本的自动化方法。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. PROJECT B. ARRAY BASED DETECTION OF ANTIGEN SPECIFIC T CELLS. Mark Davis, P.I., Yueh-hsiu Chien, and Patrick Brown and Co-P.l.s. Overview. The ability to detect and quantitate antigen specific T cells in human peripheral blood with the tetrameric peptide/MHC staining reagents have proven to be very useful for following disease progression and for evaluating the efficacy of intervention. However, this technique requires a fluorescent activated cell sorter, which severely limits the throughput, the number of specificities that can be surveyed and the potential for automation. In addition, it requires relatively large quantities of blood, which is a particular problem with samples from young children (as noted in the preceding proposals). To facilitate the analysis of immune responses developed during Influenza virus infection and to analyze the effect of vaccination, we have generated preliminary results showing that the tetramers can be arrayed on glass slides to retain antigen specific T cells and the retained T cells can be detected with fluorescent microscopy. The goal of this application is to expand and refine these results to develop a high throughput, array-based, antigen specific T cell quantitation assay that would allow the assessment of many different T cell specificities with a single blood sample. This should provide a convenient and potentially automated means of surveying multiple samples.