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. 2010 Aug 30;207(9):1863–1870. doi: 10.1084/jem.20092514

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© 2010 Kim et al.

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Figure 4. — SpA_KKAA immunization modifies host immune responses to staphylococcal infection. (A) Human or mouse serum antibody titers to diphtheria toxoid (CRM197) and nontoxigenic SpA_KKAA or SpA-D_KKAA. Human volunteers with a history of diphtheria (DTaP) immunization and staphylococcal infection (n = 16), as well as mice (n = 20) that had been infected with S. aureus Newman or USA 300 LAC or immunized with SpA_KKAA or SpA-D_KKAA, were examined by quantitative immunoblot. (B) Cohorts of BALB/c mice (n = 15) were immunized with SpA_KKAA or with PBS/adjuvant control (mock) and then challenged by intravenous inoculation with 5 × 10⁶ CFU S. aureus USA300 LAC. 30 d after infection, animals were bled and serum samples were analyzed for antibody responses to staphylococcal antigens. 27 recombinant His₆-tagged staphylococcal proteins were purified by Ni-NTA affinity chromatography and immobilized on nitrocellulose membrane at 2 µg. Signal intensities in sera from mice were quantified and normalized by infrared imaging. Data are the means and error bars represent ±SEM. Results in A and B are representative of three independent analyses.