The Immune Monitoring Core (Core C) will be comprised of technologies and services offered by two service laboratories within the laboratories of this program project: the Lyerly/Clay lab and the Immune Reconstitution Facility (Chao Lab). Core C will routinely provide investigators with high quality, state-of-the-art flow cytometry, cellular monitoring by ELIspot, proliferation, and lysis assays, protein array, and gene expression assay support for their basic and applied research efforts. In addition to providing these services, the Core will be instrumental in developing new methodologies and making them available to the P-O1 investigators. Administrative oversight for the Core will institute a charge-back mechanism to remain revenue neutral. The five goals of the Immune Monitoring Core are: 1) Provide state-of-the-art, multi-color, fluorescence activated cell sorting and analytical support for basic or translational research conducted by the P-O1 investigators. This includes high-speed sorting and multi-color analysis of cellular subsets. Assays are available for surface and intracellular marker analysis of mouse, rat, rhesus, baboon and human cells. 2) Provide state-of-the-art ELISpot, ELISA, proliferation and lysis assays for the functional characterization of antigen-specific T cell responses and anti-viral immunity. 3) Provide targeted multiplex protein array profiling of mouse and human samples, using our BioPlex bead array reader (BioRad) and Luminex bead technology. 4) Provide quantitative real-time-PCR multiplex gene expression analysis to investigators. Specific assays include T cell receptor excision circle quantification in mice and humans (qPCR), and simultaneous quantification of up to four target mRNA species in a single sample (qRT-PCR). 5) Provide state-of-the-art patient sample handling and assessment of immune responses, with standardized SOPs, quality assurance and quality control practices to monitor assay performance and validate data quality. By educating and working in collaboration with the P-O1 scientific program users, we will develop and optimize their specific assays and effectively utilize the state-of-the-art instrumentation offered by the Core.
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