Production of IL-12 and IL-10 by mature DCs in patients with CD40 deficiency. (A-B) Comparison of IL-12 and IL-10 production by mature DCs in patients with CD40 deficiency. Monocyte-derived immature DCs (2 × 105 cells per well) were cultured for an additional 48 hours with either CD154 trimer (light gray bars), LPS + IFN-γ (dark gray bars), or medium alone (black bars), as described in “Patients, materials, and methods.” IL-12 (A) and IL-10 (B) concentrations in supernatants obtained after 48 hours of culture were evaluated by ELISA. Data are expressed in pg/mL (mean ± SD of triplicates). *Significant difference in patients compared with a healthy subject as assessed by statistical analysis (P < .05). (C) Production of IL-12 in the presence or absence of IL-10-neutralizing antibody by mature DCs from a healthy subject and from HIGM3 patient 1. Monocyte-derived DCs (2 × 105 cells per well) from a healthy subject and from patient 1 were matured with soluble TNF-α or LPS + IFN-γ 48-hour stimulation (see “Patients, materials, and methods”) in the presence of anti-IL-10 (10 μg/mL) to neutralize IL-10 action on the cells (black bars). An IgG antibody was used in control experiments (light gray bars). In the supernatant of these cultures, the cytokine IL-12 was measured using ELISA, and data are expressed in pg/mL (mean ± SD of triplicates). These experiments were performed in 2 different laboratories. Data shown are from 1 of 3 representative, independent experiments.
