Laboratory Sciences Core (Core D)

实验室科学核心-项目摘要 CFAR实验室科学（LSC）提供专门设计的服务，以满足艾滋病毒临床， 翻译和基础科学研究人员。在过去的补助金周期中，我们为 CFAR调查员，具有易于访问的调度和计费系统，无论调查员的 实验室经验或培训水平。这些包括细胞分离和冷冻保存方案， PBMC、脂肪组织和皮肤。我们开发了用于分析流式细胞术的细胞染色方案，以评估 B和T淋巴细胞表型和功能，以及流式分选分析，以大量分选 淋巴细胞，或进行单细胞分选以进行详细的转录组学分析。范德比尔特长期以来 为共享核心设施提供机构支持，并进行相应投资。LSC建立的服务 经过精心设计，以满足其他非CFAR范德比尔特核心项目未能满足的需求 设施，同时利用同样令人印象深刻的机构核心， 提供给范德比尔特和梅哈里的调查人员。我们的成功使我们能够预见未来 这是CFAR的需要。一个机构核心，Vantage（范德比尔特先进基因组技术）， 显著增强了我们分析单细胞的能力，包括B细胞受体和T细胞受体测序， 和RNA转录谱。Vantage还设计了检测整个HIV基因组的方法，包括 整合位点，测序。除了提供自己的服务外，LSC还将发挥礼宾服务的作用 的服务.我们将帮助研究人员设计适当的流式细胞仪面板，并对细胞进行分类， 进行测序和分析的Vantage。LSC的使命是继续提供可持续的 领导和服务专门响应艾滋病毒调查人员的需求，使这些服务得到利用 没有CFAR是无法想象的LSC将与CFAR研究人员密切合作，设计检测方法， 促进分析性能，并帮助解释结果。LSC将促进基础，临床和转化 通过两个具体目标进行艾滋病毒研究：1）提供专业知识、咨询、检测性能和培训 包括那些需要BSL 3程序的免疫学技术。其中包括研究设计和 实施最先进的测定法，以测量与HIV相关的细胞亚群的表型和功能， 定量血浆样品和体外培养物中的HIV蛋白和核酸。2)提供礼宾服务 与艾滋病毒相关的基因组学研究服务。服务包括获得单细胞转录组学 技术、病毒基因组进化序列分析和与HIV相关的宿主微生物组分析 发病机理

Laboratory Sciences Core – PROJECT SUMMARY The CFAR Laboratory Sciences (LSC) provides services specifically designed to meet the needs of HIV clinical, translational and basic science researchers. Over the past grant cycle, we established multiple services for CFAR investigators, with an easily accessible scheduling and chargeback system, regardless of investigators' prior level of laboratory experience or training. These include cell isolation and cryopreservation protocols for PBMCs, adipose tissue, and skin. We developed cell staining protocols for analytical flow cytometry to evaluate the phenotype and function of B and T lymphocytes, and flow sorting assays to bulk sort populations of lymphocytes, or to perform single-cell sorting for detailed transcriptomic analyses. Vanderbilt has long prioritized institutional support for shared core facilities and invested accordingly. The services that the LSC established over the prior funding cycle were carefully designed to address needs unmet by other non-CFAR Vanderbilt core facilities, while simultaneously taking advantage of the impressive range of institutional cores that are equally available to investigators at Vanderbilt and Meharry. Our successes have allowed us to anticipate the future needs of this CFAR. An institutional core, VANTAGE (VANderbilt Technologies for Advanced Genomics), has significantly enhanced our ability to analyze single cells, including B cell receptor and T cell receptor sequencing, and RNA transcriptional profiling. VANTAGE has also designed assays to perform whole HIV genome, including integration site, sequencing. The LSC, in addition to providing its own services, will function as a concierge service. We will help investigators design appropriate flow cytometry panels, and sort cells, and leverage VANTAGE to perform sequencing and analysis. The mission of the LSC is to continue to provide sustainable leadership and services specifically responsive to the needs of HIV investigators, so that these services are used in ways unimaginable without CFAR. The LSC will work closely with CFAR investigators to design assays, facilitate assay performance, and help interpret results. The LSC will promote basic, clinical and translational HIV research through two Specific Aims: 1) To provide expertise, consultation, assay performance, and training in immunology techniques including those that require BSL3 procedures. These include study design and implementation of state-of-the-art assays to measure phenotype and function of cell subsets relevant to HIV and to quantitate HIV protein and nucleic acid in plasma samples and in vitro culture. 2) To provide concierge level service for genomics research relevant to HIV. Services include access to single-cell transcriptomics technologies, viral genome evolutionary sequence analysis, and host microbiome analysis relevant to HIV disease pathogenesis.