HUMAN GENETIC ANALYSIS RESOURCE

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 项目B。基于阵列的抗原特异性T细胞检测。 马克·戴维斯私家侦探作者：Yueh-shiu Chien，and帕特里克布朗和Co-P.l.s. 概况.本发明提供了用免疫荧光法检测和定量人外周血中抗原特异性T细胞的能力， 四聚体肽/MHC染色试剂已被证明对于以下疾病非常有用 并评估干预的有效性。然而，这种技术需要荧光 激活的细胞分选仪，这严重限制了吞吐量，可以被识别的特异性的数量， 调查和自动化的潜力。此外，它需要相对大量的血液， 这对于来自幼儿的样本来说是一个特别的问题（如前面的建议中所指出的）。到 有助于分析流感病毒感染期间产生的免疫应答，并分析 疫苗接种的影响，我们已经产生了初步的结果表明，四聚体可以排列 以保留抗原特异性T细胞，并且保留的T细胞可以用荧光标记检测。 显微镜本申请的目标是扩展和细化这些结果以开发高通量， 基于阵列的抗原特异性T细胞定量测定，可以评估许多 不同的T细胞特异性。这将提供一个方便和潜在的 测量多个样本的自动化手段。

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.