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.