Fig. 4. Evaluation of the parasite burden.

BALB/c mice (n = 8 per group) were immunized with ChimeraT, rPHB, rEIF5a, rLiHyp1 or rLiHyp2 (15 μg each per mouse) mixed with saponin. Controls received saline or saponin alone. Thirty days after the last vaccine dose, mice were challenged with 10⁷L. infantum stationary promastigotes and, 45 days after infection, their spleens, livers, draining lymph nodes, and bone marrows were collected and the parasite load was quantified by a limiting-dilution technique. Data were expressed as the negative log of the titer adjusted per milligram of organ; the columns indicate the means and the error bars, the standard deviations. (*) indicates statistically significant difference compared to the saline and saponin groups (P < 0.0001). (**) indicates statistically significant difference compared to the rEIF5a/saponin, rLiHyp1/saponin and rLiHyp2/saponin groups (P < 0.05). 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.