Immunology Core

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The Immunology Core (Core D) is a new core that was established after discussion by the SPORE Internal and External Advisory Committees and in response to the previous critique of our renewal application. The principal goal of the Core is to establish standardized assays of cellular and humoral immune responses to the vaccines being developed and tested by the four projects of the SPORE program. Standardization will be achieved by use of uniform reagents and protocols, trained personnel and stringent quality control measures. Centralized performance of these assays will facilitate head-to-head comparisons of vaccines that use the same primary immunological outcome measures and provide for efficient use of resources. Because some SPORE investigators have a financial interest in the vaccines that are being tested, assessment of immunogenicity by an independently directed laboratory will also allay concerns over possible conflicts of interest. The services provided by the Core to the individual projects are the following. For Project I, the assays include serum IgG-specific HPV 16 L1 virus like particle (VLP) enzyme linked immunosorbent assay (ELISA) and HPV 16, 31 and 33 in vitro pseudovirion neutralization assays. The latter 2 types are included to assess possible cross neutralization of genetically related types. For Project II, the Core will perform an ELISA to detect serum IgG directed against the vaccinogen, HPV L2 11-200x3 protein and in vitro pseudovirion neutralization assays for HPV 6 and 11 and 15 high risk HPV types. For the subset of subjects vaccinated with Gardasil, serum samples will be tested in the HPV 16 VLP ELISA and HPV 6, 11, 16, 18, 31, 33 and 45 pseudovirion neutralization assays. For Projects III and IV, the Core will perform IFN- gamma ELISPOT assays on unfractionated peripheral blood mononuclear cells (PBMC) following overnight stimulation with pools of E6 or E7 peptides. The CD8+ and CD4 + phenotype of the responding lymphocytes will be confirmed by intracellular cytokine staining and flow cytometry. Secondary immunologic assays, assays specific to an individual project and novel immunologic assay development will be performed and perfected by the investigators within the individual projects and only transferred to the Immunology Core when fully optimized and standardized. The Immunology Core will interact extensively with the individual projects as well as the Tissue/Pathology Core (Core C), which will provide specimens, and the Biostatistlcs/Data Management Core (Core B), which will perform data analyses.