Immune Profiling Core

抽象-免疫分析核心 免疫分析核心(IPC)将提供全面的技术检测支持服务 这两个构成U19提案的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、程序和运营中获得最大的综合实力 其组成部分的协同效应。支撑这些努力的是杜克免疫档案核心(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 作为人类免疫分析平台的一部分，核心积极参与外部能力测试和 超过25年的质量保证计划。化验应在良好的临床实验室中进行 符合(GCLP)的环境，符合高度标准化和经过验证的标准操作程序 (天哪)。一支由来自以下国家的经验丰富的实验室专业人员组成的支持团队加入了Core的工作 柯克和克内克特尔的研究核心最近从埃默里大学调到了杜克大学。这些 个人在人类和非人类移植研究的各个方面都有广泛的专业知识。 灵长类(NHP)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 标本处理和生物库，2)分析多色流式细胞术(PFC)表型，以及 细胞内细胞因子染色(ICCS)，3)病毒学检测，4)综合组织学和 免疫组织化学支持。IPC处于有利地位，可以向该计划下的项目提供PFC服务 提案；PFC一直是DIPC研究工作的技术支柱。目前的核心 该设施配备了4激光器BD LSRII、5激光器BD LSRFortessa、5激光器BD LSR-X20，以及最近的5激光器 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个参数的流式细胞仪。提供全面的免疫学 为了提供支持，我们为IPC实现了几个高度开发的组件。一次样品加工和 在柯克博士的领导下开始于埃默里的生物库服务已重新建立为底物服务和 在杜克就任外科主任后，他在杜克大学获得了研究支持服务(SSCRSS)。 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，将允许IPC 为了监测NHP中与临床相关性最强的三种潜伏病毒：恒河猴巨细胞病毒(RhCMV)， 恒河猴淋巴病毒(RhLCV)(与EB病毒同源)和猿猴病毒40(SV40)。IPC将 通过我们的免疫组织化学服务提供常规的组织学和免疫组织化学分析 实验室，这将允许每个项目检查实验操作对关键主机的影响 组织。将特别强调检查生发中心形成的影响之后的各种 诱导疗法。人力和NHP专业知识的并列代表了这一IPC的特殊优势 财团，进一步促进拟议研究的高度翻译性。

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.