Laboratory Sciences Core (Core D)

实验室科学核心-项目摘要 CFAR实验室科学(LSC)提供专门为满足艾滋病毒临床需求而设计的服务， 翻译和基础科学研究人员。在过去的赠款周期中，我们为以下项目建立了多项服务 CFAR调查人员，具有易于访问的调度和按存储容量使用计费系统，无论调查人员的 先前的实验室经验或培训水平。这些包括细胞分离和冷冻保存协议，用于 外周血单核细胞、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来评估 B和T淋巴细胞的表型和功能，以及批量分选人群的流动分选试验 淋巴细胞，或执行单细胞分选以进行详细的转录分析。范德比尔特长期以来一直优先考虑 为共享的核心设施提供机构支持，并进行相应的投资。LSC建立的服务 在前一个供资周期内，精心设计以满足其他非CFAR范德比尔特核心项目没有满足的需求 设施，同时利用令人印象深刻的一系列机构核心 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 这项CFAR的需求。机构核心Vantage(Vanderbilt Technologies For Advanced Genome)拥有 显著增强了我们分析单个细胞的能力，包括B细胞受体和T细胞受体测序， 和RNA转录图谱。Vantage还设计了进行艾滋病毒全基因组检测的方法，包括 整合地点，测序。LSC除了提供自己的服务外，还将发挥礼宾的作用 服务。我们将帮助调查人员设计合适的流式细胞仪面板，并对细胞进行分类，并利用 进行测序和分析的优势。LSC的使命是继续提供可持续的 具体回应艾滋病毒调查人员需求的领导力和服务，以便使用这些服务 在没有CFAR的情况下，这种方式是不可想象的。LSC将与CFAR调查人员密切合作，设计检测方法， 促进化验性能，并帮助解释结果。LSC将促进基础、临床和翻译 通过两个具体目标进行艾滋病毒研究：1)提供专门知识、咨询、化验业绩和培训 在免疫技术方面，包括那些需要BSL3程序的技术。这些包括研究设计和 实施最先进的检测方法来测量与艾滋病毒和艾滋病相关的细胞亚群的表型和功能 对血浆标本和体外培养中的HIV蛋白和核酸进行定量。2)提供礼宾级别 为与艾滋病毒相关的基因组研究提供服务。服务包括获得单细胞转录本 与HIV相关的技术、病毒基因组进化序列分析和宿主微生物组分析 疾病发病机制。

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.