Automated comparison of flow data from HIV and vaccine infected subjects.

描述（由申请人提供）：我们的实验室构思并开发了最初的流式细胞仪仪器，此后引入了许多创新，增加了该技术的广泛实用性和可访问性。 作为这项工作的一部分，我们开发了关键的软件和分析能力，现在在流技术的研究和临床用户中无处不在，其中艾滋病毒研究人员和临床医生显然占主导地位。 在这里的研究中，我们建议利用我们长期的流动专业知识，引入急需的流动能力，以促进高正常和高通量的工作HIV试验样品和疫苗开发期间分析的样品 基本上，我们建议开发创新的方法来自动化流数据的数据处理和处理任务。 这将包括我们最近发表的用于定位和识别感兴趣的子集中的细胞的方法的应用，以便我们可以对这些子集中/上的子集标志物表达进行统计学上稳健的比较。 这些方法还将使我们能够开发创新方法，用于识别由抗原刺激或来自试验和/或其他受试者的冷冻或新鲜收集的样品的其他刺激的标记物变化的个体或组。 重要的是，该软件旨在（首次）提供疫苗开发模型中体内或离体刺激的T细胞亚群中诱导的几种细胞内细胞因子表达的非协调变化的稳健单一测量。 由于其设计，我们建议以集成格式创建我们为这些任务构建的软件，该软件易于自动化，以实现高通量分析。 公共卫生相关性：我们建议开发创新的方法来自动化关键处理和数据处理任务，这些任务需要直接和自动分析从HIV研究和疫苗试验中收集的样本的流式细胞术数据。一旦建立，该软件也将对艾滋病毒临床实践有用。 重要的是，我们提出的工作将引入全新的基于免疫学的方法来识别T细胞亚群，并应用全新的统计方法来比较疫苗试验和其他HIV研究样品刺激诱导的亚群标志物表达的变化。 与所有当前方法不同，这些新方法将允许计算对刺激的响应的单个指数，即使当测量的响应需要整合T细胞亚群中或T细胞亚群上的几个标记物发生的数据不协调变化时。

DESCRIPTION (provided by applicant): Our laboratory conceived and developed the initial flow cytometry instruments and has since introduced many innovations that increase the broad utility and accessibility of this technology. As part of this effort, we developed key software an analysis capabilities that are now ubiquitous among research and clinical users of flow technology, among whom HIV researchers and clinicians clearly predominate. In studies here, we propose to exploit our long-term flow expertise in the interests of introducing much needed flow capabilities to facilitate high normal and high-throughput work HIV trial samples and samples analyzed during vaccine development Basically, we propose to develop innovative methods to automate data handling and processing tasks for flow data. This will include the application of our recently published methods for locating and identifying cells in subsets of interest, so that we can do statistically robust comparisons of subset marker expression in/on these subsets. These methods will also allow us to develop innovative methods for identifying individual or groups of marker changes stimulated by antigen or other stimulation of frozen or freshly collected samples from trial and/or other subjects. Importantly, this software is intended to provide (for the first time) a robust single measure of non-coordinated changes in expression of several intracellular cytokines induced in T cell subsets stimulated in vivo or ex vivo in vaccine development models. Because of its design, we propose to create the software we build for these tasks in an integrated format that is readily amenable to automation to enable high-throughput analyses. PUBLIC HEALTH RELEVANCE: We propose to develop innovative methods to automate key handling and data processing tasks needed for direct and automated analysis of flow cytometry data collected for samples from HIV studies and vaccine trial. Once established, this software will also be useful for HIV clinical practice. Importantly, the work we propose will introduce entirely new statistics-based methods for identification of T cell subsets and the application of entirely new statistical methods to compare changes in subset marker expression induced by stimulation of samples from vaccine trials and other HIV studies. Unlike all current methods, these new methods will allow computation of a single index of response to stimulation, even when the response being measured requires integration of data uncoordinated changes that occur for several markers in or on T cell subsets.