Figure 3
Treatment of L. infantum axenic amastigote-infected mice with reference drugs miltefosine and amphotericin B. (A) Schematic representation of the experimental design. BALB/c mice were infected with 108 luciferase-expressing L. infantum axenic amastigotes (AxAMA) IV and 4-day treatments with either 20 mg/kg/day of miltefosine per os (PO) or 1 mg/kg/day of Amphotericin B (IV) were initiated 15 days post-infection (DPI). All animals (n = 4 per group) were imaged right before (day 15 post-infection), one day after (day 19 post-infection) and 3 days (day 21 post-infection) after the end of treatment using the IVIS Lumina LT system. At the last time point animals were sacrificed and parasite burden in the liver, spleen, and femur bone marrow were determined by limiting dilution. (B) Images of infected mice resulting from the superimposition of the bioluminescence signal map and a grey-scale photograph of the mice. The ROIs shown were used to quantify the bioluminescence signal originating from the liver and spleen anatomical regions. (C) Bioluminescence measurements expressed as average radiance (photons/s/cm2/steradian) corresponding to the previously defined liver (graph on the left) and spleen (graph on the right) ROIs. Means ± standard deviations are represented in bars. The dotted line represents the background signal calculated by applying the ROIs on images of uninfected animals. Statistical significance calculated by two-way ANOVA: (*)p < 0.05, (**)p < 0.01, (****)p < 0.0001. (D) Parasite burden in the liver (graph on the left) and spleen (graph on the right) determined by limiting dilution 21 days post-infection. The dotted lines represent the upper and lower detection limit of the technique for each organ. Means ± standard deviations are represented in bars. Statistical significance calculated by ordinary one-way ANOVA: (*)p < 0.05, (***)p < 0.001. Data representative of two independent experiments.
