Production of cytokines by spleen cells of vaccinated mice after Leishmania infection . Mice (four per group) immunized with vaccine diluent (Saline), with CpG-ODN adjuvant (CpG) or with the recombinant vaccine (LmL3 + LmL5 + CpG) were independently challenged subcutaneously in the left footpad with 1 × 106 stationary- phase promastigotes of L. amazonensis (A) or 1 × 107 stationary-phase promastigotes of L. chagasi (B). Nine weeks after infection, spleen cell cultured suspensions were non-stimulated (Medium; background control) or separately stimulated with recombinant LmL3 or LmL5 (12 μg/ml each one) or with SLA from L. amazonensis (LaSLA; A) or L. chagasi (LcSLA, B) at 12 μg/ml each one for 48 h at 37°C, 5% CO2. IFN-γ , IL-4 and IL-10 levels were measured in culture supernatants by ELISA. Mean + standard deviation (SD) of cytokine levels determined in four individual mice per group is shown; *, P < 0.05 indicates significant cytokine production increase between LmL3 + LmL5 + CpG-ODN vaccinated mice and both control groups; +, P < 0.05 indicates significant cytokine production decrease between LmL3 + LmL5 + CpG-ODN vaccinated mice and both control groups. Data shown are representative of two independent experiments with similar results.