Fig. 6: Cytokine production after L. infantum challenge.
BALB/c mice (n = 8 per group) immunized with ChimeraT, rPHB, rEIF5a, rLiHyp1 or rLiHyp2 (15 μg each per mouse) plus saponin were challenged with 107 L. infantum stationary promastigotes and, 45 days after infection, their spleens were collected. Spleen cells (5 × 106 per mL) were cultured in DMEM (medium, control) or stimulated with each immunizing protein (10 µg/mL each) or L. infantum SLA (50 µg/mL) for 48 h at 37 °C with 5% (v/v) CO2. a Measurement of IFN-γ, IL-12, GM-CSF, IL-4, and IL-10 cytokine levels in cell supernatants. b Ratios between the IFN-γ and IL-10 levels. c Evaluation of IFN-γ, IL-12, GM-CSF, IL-4, and IL-10 production in spleen cells from the Chimera T/saponin-immunized mice stimulated in vitro with the synthetic peptide epitopes. The bars indicate the mean and the error bars denote the standard deviation of the groups. (*) indicates significant difference in relation to the groups of mice immunized with the recombinant proteins plus adjuvant and challenged with L. infantum parasites (P < 0.005). (**) indicates significant difference in relation to using the individual peptides as stimuli (P < 0.01). (+) indicates significant difference in relation to the groups of mice immunized with the Chimera T or recombinant proteins plus adjuvant and challenged with L. infantum parasites (P < 0.001). Statistics were done using one-way analysis of variance (ANOVA), followed by Bonferroni´s post-test, which was used for multiple comparisons, and P values < 0.05 were considered significant.
