Figure 3. Optimal dosage for adjuvanted vaccines.
BALB/c mice (5 per group) were vaccinated once with the indicated dose of Ad5-Gag. Panel D and Panel E: Equal concentrations of AD5-Gag & SP-D-CD27L or irrelevant protein (GFP) virus were added where specified. 2×105 fresh splenocytes were cultured in a 96 well MAIP plate coated with IFN-γ (Panel A and Panel C), IL-4 (Panel B), or IL-2 capture antibody (Panel D), in the presence of 5 ug/ml HIV-1 Gag peptide AMQMLKETI. After 18 hours plates were developed with AEC substrate and counted in an automated plate reader. Data displayed as Spot-Forming Cells (SFC) per million splenocytes. Panel C: Anti-Gag IgG ELISA. Before sacrifice, mice were bled and serum was isolated. Serum at indicated dilution was cultured for 2 hours on ELISA plates coated with HIV-1 p55 Gag protein (10 µg/ml). Plates were washed and probed with alkaline phosphatase-conjugated goat anti-mouse IgG antibody for 1 hour before developing with BluePhos substrate. Absorbance was measured at 650 nm.