Immune Profiling Core

免疫分析核心（IPC）将为包含此U19提案的两个项目中提出的假设驱动的研究提供全面的技术分析支持服务。核心从相关的专业知识、组织、程序和其组成部分的操作协同中获得最大的整体优势。支撑这些努力的是杜克大学免疫分析核心（DIPC），这是杜克大学医学院自1988年以来一直存在的共享资源。核心主要专注于基于流式细胞仪的人类免疫分析平台，积极参与外部能力测试和质量保证计划超过25年。根据高度标准化和经过验证的标准操作程序（sop），在符合良好临床实验室（GCLP）标准的环境中进行分析测试。核心是由一个经验丰富的实验室专业人员组成的支持团队加入的，他们来自Kirk和Knechtle研究核心，他们最近从埃默里大学搬迁到杜克大学。这些人在移植研究的各个方面都有广泛的专业知识，包括人类和非人灵长类动物（NHP）的研究。IPC将为项目1和项目2提供的主要服务包括：1)最先进的标本处理和生物银行，2)分析多色流式细胞术（PFC）表型分析和细胞内细胞因子染色（ICCS）， 3)病毒学测试，以及4)全面的组织学和免疫组织化学支持。IPC完全有能力为该提案下的项目提供PFC服务；PFC一直是DIPC研究工作的技术支柱。目前的核心设施包括4激光BD LSRII， 5激光BD LSRFortessa， 5激光BD LSR-X20和最近的5激光BD LSR-X50。在这些数字仪器中，前3台能够进行多达18-24个参数的单细胞分析，而X50能够支持50个参数的流式细胞术。为了提供全面的免疫学支持，我们为IPC实施了几个高度开发的组件。在Kirk博士担任杜克大学外科系主任后，Emory大学开始的标本处理和生物库服务被重新建立为基质服务和研究支持服务（SSCRSS）。通过Kirk博士实验室的工作，我们还建立了一个病毒检测设施，使IPC能够监测NHPs中最具临床相关性的三种潜伏病毒：恒河巨细胞病毒（rhCMV）、恒河淋巴隐病毒（rhLCV）（恒河与EBV同源）和猿猴病毒40 （SV40）。IPC将通过我们的免疫组织化学服务实验室提供常规组织学和免疫组织化学分析，这将允许每个项目检查实验操作对关键宿主组织的影响。特别强调的是检查各种诱导方案对生发中心形成的影响。人类和非传染性疾病专业知识的结合代表了该IPC联盟的一个特殊优势，进一步促进了拟议研究的高度转化方面

Immune Profiling Core The Immune Profiling Core (IPC) will provide comprehensive technical assay support services for the hypothesis-driven research studies proposed in the two Projects comprising this U19 proposal. The Core derives its greatest overall strength from the relevant expertise, organization, procedures, and operational synergy of its components. Anchoring these efforts is the Duke Immune Profiling Core (DIPC), a Duke School of Medicine Shared Resource that has been in existence since 1988. Largely focused on flow cytometry-based platforms for human immune profiling, the Core has actively participated in external proficiency testing and quality assurance programs for over 25 years. Assay testing is performed in a Good Clinical Laboratory (GCLP)-compliant environment according to highly standardized and validated Standard Operating Procedures (SOPs). The Core is joined in its efforts by a support team of highly experienced laboratory professionals from the Kirk and Knechtle research Cores who have recently re-located to Duke from Emory University. These individuals have extensive expertise in all aspects of transplantation research, both in human and nonhuman primate (NHP) studies. The key services the IPC will provide to Projects 1 and 2 include: 1) state-of-the-art specimen processing and biobanking, 2) analytical polychromatic flow cytometry (PFC) phenotyping, and intracellular cytokine staining (ICCS), 3) virologic testing, and 4) comprehensive histology and immunohistochemistry support. The IPC is well positioned to provide PFC services to the projects under this proposal; PFC having been the technical mainstay of the research endeavors of the DIPC. The present core facility houses a 4-laser BD LSRII, 5-laser BD LSRFortessa, 5-laser BD LSR-X20 and, most recently, a 5-laser BD LSR-X50. Of these digital instruments, the first 3 are capable of up to 18-24 parameter single cell analyses, while the X50 is capable of supporting 50-parameter flow cytometry. To provide comprehensive immunologic support, we have implemented several highly developed components for the IPC. A specimen processing and biobanking service that began at Emory under Dr. Kirk has been re-established as the Substrate Services and Research Support Service (SSCRSS) at Duke following his arrival as Chairman of the Department of Surgery. Through the work of Dr. Kirk's laboratory, we have also established a viral testing facility that will allow the IPC to monitor three of the most clinically-relevant latent viruses in NHPs: Rhesus Cytomegalovirus (rhCMV), Rhesus Lymphocryptovirus (rhLCV) (rhesus homology of EBV), and Simian Virus 40 (SV40). The IPC will provide routine histology and immunohistochemistry assays through our immunohistochemistry service laboratory, which will allow each project to examine the effects of the experimental manipulations on key host tissue. Special emphasis will be placed on examining effects on germinal center formation following the various induction regimens. The juxtaposition of human and NHP expertise represents a particular strength of this IPC consortium, further facilitating the highly translational aspects of the proposed studies