Immune Profiling Core

免疫分析核心

• 批准号: 10649944
• 负责人: Kent J. Weinhold
• 金额: $ 26.01万
• 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 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10649944
• 关键词: Adjuvant Therapy; Antibodies; Antigens; Autopsy; B-Lymphocytes; Biological Assay; Biological Specimen Banks; Biopsy; Body Fluids; Cell Death; Cells; Chairperson; Clinical; Color; Communicable Diseases; Core Facility; Cryopreservation; Cytomegalovirus; Disease; Environment; Flow Cytometry; Goals; Graft Rejection; Health; Histology; Human; Human Herpesvirus 4; Immune; Immunity; Immunization; Immunoassay; Immunocompromised Host; Immunohistochemistry; Immunologics; Immunology procedure; Immunosuppression; Individual; Infrastructure; Kidney Transplantation; Knowledge; Laboratories; Lasers; Lead; Leadership; Lymphocryptovirus; Macaca mulatta; Management Information Systems; Memory B-Lymphocyte; Monitor; Operative Surgical Procedures; Organ; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Plasma; Plasma Cells; Positioning Attribute; Pre-Clinical Model; 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; Transplantation Immunology; Universities; Viral; Viral Antigens; Virus Latency; Work; allotransplant; base; biobank; clinically relevant; cytokine; desensitization; digital; exhaustion; improved; instrument; kidney allograft; laboratory experience; lymph nodes; medical schools; nonhuman primate; novel therapeutic intervention; post-transplant; preservation; prevent; programs; quality assurance; research study; secondary lymphoid organ; side effect; single cell analysis; synergism; testing services

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

免疫分析核心免疫分析核心（IPC）将为由U19提案组成的两个项目中提出的假设驱动的研究提供全面的技术测定支持服务。核心从其组件的相关专业知识，组织，程序和运营协同作用中获得了最大的总体力量。杜克免疫分析核心（DIPC）是杜克大学医学学院共享资源，自1988年以来一直存在。主要集中在基于流式细胞仪的人类免疫分析的平台上，该核心已积极参与了25年以上的外部熟练度测试和质量保证计划。根据高标准化和经过验证的标准操作程序（SOP），在良好的临床实验室（GCLP）编译环境中进行测定测试。来自柯克和诺克特尔研究核心的经验丰富的实验室专业人员的支持团队加入了核心，他们最近从埃默里大学重新分配给了杜克大学。这些人在人类和非人类灵长类动物（NHP）研究的移植研究的各个方面都有广泛的专业知识。 IPC将向项目1和2提供的关键服务包括：1）最先进的标本处理和生物群，2）分析性多色流式细胞术（PFC）表型，以及细胞内细胞因子染色（ICCS），3）病毒学测试，以及4）综合组织学和免疫学组成症。 IPC在此提案下为项目提供PFC服务的好处； PFC是DIPC研究努力的技术中流tay柱。目前的核心设施设有4射线BD LSRII，5射线BD LSRFORTESSA，5-laser BD LSR-X20，以及最近的5射线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，而X50能够支持50参数流式细胞仪。为了提供全面的免疫支持，我们为IPC实施了几个高度发展的组件。在柯克博士（Kirk）领导下的埃默里（Emory）的一项标本加工和生物群体服务已被重新建立为杜克大学（Duke）担任外科部董事长后的基板服务和研究支持服务（SSCRSS）。通过Kirk博士的实验室的工作，我们还建立了一个病毒测试设施，该设施将使IPC可以监测NHPS中最临床上最临床最重要的潜在病毒：恒河猴胞膜瘤病毒（RHCMV），瑞斯斯淋巴结病毒（rhlcv）（Rhlcv）（Rhlcv）（Rhelsus obv）和Simian Aimian病毒式40（SV）40（SV）40（SV）40（ IPC将通过我们的免疫组织化学实验室提供常规的组织学和免疫组织化学测定，这将使每个项目都可以检查实验操纵对关键宿主组织的影响。在各种诱导方案之后，将特别强调检查对生发中心形成的影响。人类和NHP专业知识的并置代表了该IPC财团的特殊优势，进一步促进了拟议的研究的高度转化方面