Immune Profiling Core

免疫分析核心

• 批准号: 9980787
• 负责人: Kent J. Weinhold
• 金额: $ 46.12万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980787
• 关键词: Adjuvant Therapy; Antibodies; Antigens; Autopsy; B-Lymphocytes; Biological Assay; Biological Specimen Banks; Biopsy; Body Fluids; Catalogs; Cell Death; Cells; Chairperson; Clinical; Color; Communicable Diseases; Core Facility; Cryopreservation; Cytomegalovirus; Environment; Flow Cytometry; Health Services Research; Histology; Human; Human Herpesvirus 4; Immune; Immunization; Immunoassay; Immunocompromised Host; Immunohistochemistry; Immunologics; Immunology; Immunology procedure; Individual; Infrastructure; Kidney Transplantation; Knowledge; Laboratories; Lasers; Lead; Leadership; Lymphocryptovirus; Macaca mulatta; Management Information Systems; Memory B-Lymphocyte; Monitor; Operative Surgical Procedures; Peripheral Blood Mononuclear Cell; Phenotype; Plasma; Plasma Cells; Positioning Attribute; Prior Therapy; Procedures; Proteasome Inhibition; Regimen; Regulation; Research; Research Support; Resource Sharing; Rhesus; Risk; Safety; Sampling; Scientist; Serum; Services; Simian virus 40; Sirolimus; Specimen; Specimen Handling; Stains; Standardization; Structure of germinal center of lymph node; Supervision; T-Lymphocyte; Technical Expertise; Testing; Thymus Gland; Tissue Sample; Tissues; Transplantation; Universities; Viral; Viral Antigens; Virus Latency; Work; base; biobank; clinically relevant; cytokine; desensitization; digital; exhaustion; instrument; kidney allograft; laboratory experience; lymph nodes; medical schools; nonhuman primate; post-transplant; prevent; programs; quality assurance; research study; secondary lymphoid organ; single cell analysis; synergism; testing services; virology

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.

摘要 – 免疫分析核心 免疫分析核心（IPC）将为患者提供全面的技术检测支持服务 构成该 U19 提案的两个项目中提出的假设驱动的研究。核心 最大的综合实力来自于相关的专业知识、组织、程序和运营 其组成部分的协同作用。杜克大学免疫分析核心 (DIPC) 是这些努力的基础 自 1988 年以来一直存在的医学共享资源。主要集中于基于流式细胞术的 人体免疫分析平台，核心积极参与外部能力验证和 质量保证计划已超过 25 年。化验测试在良好的临床实验室中进行 根据高度标准化和经过验证的标准操作程序，符合 (GCLP) 环境 （标准操作程序）。核心团队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共同努力： Kirk 和 Knechtle 研究核心最近从埃默里大学迁至杜克大学。这些 个人在人类和非人类移植研究的各个方面都拥有广泛的专业知识 灵长类动物（NHP）研究。 IPC 将为项目 1 和 2 提供的关键服务包括： 1) 最先进的 样本处理和生物样本库，2) 分析多色流式细胞术 (PFC) 表型分析，以及 细胞内细胞因子染色 (ICCS)，3) 病毒学检测，以及 4) 综合组织学和 免疫组织化学支持。 IPC 完全有能力为该项目下的项目提供 PFC 服务 提议; PFC 一直是 DIPC 研究工作的技术支柱。现在的核心 该设施拥有 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则能够支持50参数流式细胞术。提供全面的免疫学 支持，我们已经为 IPC 实现了几个高度开发的组件。标本处理及 柯克博士在埃默里大学开始的生物样本库服务已重新建立为 Substrate Services 和 在他担任杜克大学外科系主任后，他开始在杜克大学研究支持服务中心 (SSCRSS)。 通过 Kirk 博士实验室的工作，我们还建立了病毒检测设施，使 IPC 能够 监测 NHP 中与临床最相关的三种潜伏病毒：恒河猴巨细胞病毒 (rhCMV)、 恒河猴淋巴密码病毒 (rhLCV)（EBV 的恒河猴同源物）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。将特别重点检查各种方法对生发中心形成的影响。 诱导方案。人类和 NHP 专业知识的结合代表了该 IPC 的特殊优势 联盟，进一步促进拟议研究的高度转化方面。