DC generation and infection

Generation of bone marrow-derived cells (BMDCs) was performed as previously described (Carregaro et al., 2008). For the DC activation assay, BMDCs (1 × 10⁶ cells/mL) cultured in RPMI-1640 supplemented with 10% FBS were infected with L. infantum promastigotes at a 1:5 ratio (cells/parasites) for 24 h. The supernatants were collected to measure IL-12p40 (BD Biosciences, catalog number 555165) and IL-10 (R&D Systems, catalog number DY417-05) production by an ELISA kit, according to the manufacturer's instructions. BMDCs were harvested from some cultures, and their surface expression was characterized by flow cytometry using antibodies against CD11c, TLR2, MHCII, CD40, and CD86 conjugated to PECy7, PE, FITC, PerCP, APC, respectively, as well as control isotypes.