To quantify antigen-specific T cell responses, 96-well plates were coated with anti–IL-17 or anti-IFNγ antibody (Becton Dickinson Biosciences), and 5 × 105 splenocytes from infected or vaccinated mice were added to each well. The splenocytes were incubated with purified antigen or peptide (1 μg/ml) or heat-killed S. aureus (5 × 105 CFU per well) for 24 hours at 37°C. Following washing, biotin-labeled detection antibodies were added to the wells, followed by avidin–horseradish peroxidase substrate (eBioscience). Spots were counted following addition of the substrate solution (BD Biosciences) using an Immunospot series 1 analyzer (Cellular Technology).

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