Figure 4. Parasitophorous vacuole analysis.

Photomicrographs of macrophages from F₁ mice, infected with DCL (BA276) and LCL (BA125) isolates after 72 h of culture (A). Arrows point to individual PVs. Magnification 400X. The average sizes of the PVs induced by different isolates from patients with LCL (BA69, BA73 and BA 125) (□) and DCL (BA276, BA336, and BA700) (▪) after 72 h of infection were measured using Image-Pro Plus 6.0 (B). Data are shown as the area in μm² of PVs in each tested isolate. Boxes represent median values and interquartile interval of PV sizes pooled from different isolates mentioned above. The correlation between PS exposure at 24 h and PV area showed in (C). The three lower points in X axis (PS exposure) represents LCL isolates while the three higher points are from DCL isolates. Differences were checked using unpaired t test. Spearman test was used to verify the significance in the correlations between PV area and PS exposure. The r value is plotted in the correlation graph. PS exposure on the L. amazonensis amastigotes surface from nude mice (Figures D–F). Mean fluorescence intensity of annexin V staining on lesion-derived amastigotes (LV79) purified from BALB/c WT or BALB/c ^nu/nu mice (D). Graph corresponds to one representative experiment out of three. H&E staining of histological slides of infected footpads obtained from BALB/c WT or BALB/c nude mice 5 weeks post infection (E). Peritoneal macrophages derived from BALB/c mice were infected with lesion-derived amastigotes (LV79) purified from BALB/c WT or BALB/c ^nu/nu mice 5 weeks post infection. After 24 h of infection cells were fixed and stained. The infectivity index (F) was defined by microscopic analysis. Representative experiment of two repeats. Differences were checked using unpaired t test.