PCMC_IFN-γ/IL-4 activation after antigen-specific interaction with OT-II T cells. (A-B) PCMC_IFN-γ/IL-4s either unpulsed or pulsed with 10 μg/mL OVAp were cocultured with OT-II T cells in the presence of anti-DNP IgE (1 μg/mL) for 16 hours. (A) Ability to release β-hexosaminidase in response to increasing concentrations of DNP-HSA. Data are from 1 representative experiment of 4 performed in triplicates. (B) EC₅₀ (half maximal effective concentration) of DNP-HSA from 4 independent experiments. Difference between groups was evaluated by a paired Student t test by using GraphPad Prism software. (C) PCMCs_IFN-γ/IL-4 either unpulsed or pulsed with 10 μg/mL OVAp were cocultured with OT-II T cells for 16 hours. In parallel samples PCMCs were treated for 16 hours with supernatant from activated OT-II (SNT; 50%, from OT-II cultured for 6 days with anti-CD3/anti-CD28–coated beads) or medium alone. Expression of CD69 on PCMCs was analyzed by flow cytometry; numbers indicate MFI. Data are from 1 representative experiment of 3.