IL-12 p70 and IFN-γ detection by ELISA.

Ly5.2⁺CD8α⁺Ia⁺ DCs from Ly5.2⁺CD8α⁺CD11c⁻Lin⁻precursor–reconstituted Ly5.1 B6 mice (donor-derived CD8α⁺ DCs) and host-type CD8α⁺ DCs from Ly5.1 BM cell–reconstituted Ly5.1 B6 mice (host-type CD8α⁺ DCs) were purified as described in “Materials and methods,” respectively. (A) Cells (7.5 × 10⁴) from each population were cultured in the presence of GM-CSF (20 ng/mL) + IFN-γ (20 ng/mL) + Pansorbin (50 μg/mL) in 200 μL medium for 40 hours. (B) Cells (3.3 × 10⁴) from each population were stimulated in vitro with rmIL-12 (16 ng/mL) in 200 μL medium for 48 hours. Supernatants were assayed for IL-12 p70 and IFN-γ with ELISA, respectively. Results are expressed as the mean ± 1 SD of the triplicate cultures and are representative of 3 independent experiments.