Immune Profiling Core

摘要-免疫分析核心 免疫分析核心（IPC）将为免疫分析提供全面的技术分析支持服务。 在构成U19提案的两个项目中提出的假设驱动的研究。核心 从相关的专业知识、组织、程序和操作中获得最大的综合实力 其组成部分的协同作用。支持这些努力的是杜克免疫分析核心（DIPC），一个杜克学校 自1988年以来一直存在的医学共享资源。主要集中在基于流式细胞术的 作为人类免疫分析的平台，核心积极参与外部能力测试， 25年以上的质量保证体系。在药物临床试验质量管理规范（GLP）实验室进行含量测定 （GCLP）合规环境，符合高度标准化和经验证的标准操作程序 （标准操作规程）。核心是加入其努力由一个支持团队的经验丰富的实验室专业人员从 柯克和克奈希特研究中心最近从埃默里大学搬到了杜克大学。这些 个人在移植研究的各个方面都有广泛的专业知识，包括人类和非人类的 灵长类动物（NHP）研究。IPC将为项目1和2提供的主要服务包括：1）最先进的 标本处理和生物库，2）分析型多色流式细胞术（PFC）表型分析，以及 细胞内细胞因子染色（ICCS），3）病毒学检测，和4）全面的组织学和 免疫组织化学支持。IPC完全有能力为本项目下的项目提供PFC服务。 PFC一直是DIPC研究工作的技术支柱。本核心 该设施拥有4激光BD LSRII，5激光BD LSRFortessa，5激光BD LSR-X20，以及最近的5激光 BD LSR-X50。在这些数字仪器中，前3种能够进行多达18-24个参数的单细胞分析， 而X50能够支持50参数流式细胞术。提供全面的免疫学 支持，我们已经实现了几个高度发达的IPC组件。一种标本处理和 柯克博士在埃默里大学开始的生物库服务已被重新建立为基质服务， 研究支持服务（SSCRSS）在杜克后，他作为外科系主任抵达。 通过柯克博士实验室的工作，我们还建立了一个病毒测试设施， 监测NHP中三种最具临床相关性的潜伏病毒：恒河猴巨细胞病毒（rhCMV）， 恒河猴淋巴隐病毒（rhLCV）（EBV的恒河猴同源性）和猿猴病毒40（SV 40）。IPC将 通过我们的免疫组织化学服务提供常规组织学和免疫组织化学检测 实验室，这将使每个项目能够检查实验操作对关键宿主的影响， 组织.特别强调将放在检查影响germinal中心形成以下各种 诱导方案。人力和NHP专业知识的并列代表了IPC的特殊优势 财团，进一步促进拟议研究的高度转化方面。

ABSTRACT – 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.