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 假设驱动的研究在完成了U19提案的两个项目中提出的研究。核心 从相关的专业知识，组织，程序和运营中获得最大的总体优势 其组件的协同作用。锚定这些努力的是杜克大学杜克大学免疫分析核心（DIPC） 自1988年以来一直存在的医学共享资源。 人类免疫分析平台，核心已经积极参与外部能力测试和 质量保证计划超过25年。测定测试是在良好的临床实验室中进行的 （GCLP） - 根据高标准化和经过验证的标准操作程序重编的环境 （SOP）。由经验丰富的实验室专业人员的支持团队加入了核心的努力 最近从埃默里大学（Emory University）重新安置到杜克大学（Duke）的柯克（Kirk）和克尼克（Knechtle）研究核心。这些 个人在人类和非人类的移植研究的各个方面都有广泛的专业知识 灵长类动物（NHP）研究。 IPC将向项目1和2提供的关键服务包括：1） 标本加工和生物群，2）分析多色流式细胞仪（PFC）表型和 细胞内细胞因子染色（ICC），3）病毒学测试，4）综合组织学和 免疫组织化学支持。 IPC的位置很好，可以为此提供PFC服务 提议; PFC是DIPC研究努力的技术中流tay柱。现在的核心 设施设有4射线BD LSRII，5射线BD LSRFORTESSA，5-LASER BD LSR-X20，以及最近的5-laser BD LSR-X50。在这些数字仪器中，前3个最多能够进行18-24个参数单细胞分析， X50能够支持50参数流式细胞仪。提供全面的免疫学 支持，我们已经为IPC实施了几个高度开发的组件。标本处理和 Kirk博士从Emory开始的生物库服务已被重新建立为基板服务，并且 杜克大学（Duke）的研究支持服务（SSCRSS）在他担任外科部主席后。 通过柯克博士的实验室的工作，我们还建立了一个病毒测试设施，将允许IPC 监测NHP中三种最临床中最相关的潜在病毒：恒河猴（RHCMV）， 恒河-淋巴结病毒（Rhlcv）（EBV的恒河类同源性）和猿猴病毒40（SV40）。 IPC会 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测定 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。特殊重点将放在检查对各种生发中心形成的影响之上 诱导方案。人类和NHP专业知识的并置代表了此IPC的特殊优势 财团，进一步促进了拟议研究的高度翻译方面。