The Ecto-5nucleotidase/CD73 Mediates Leishmania amazonensis Survival in Macrophages

Figure 4

CD73 activity determines survival of L. amazonensis. Resident macrophages were obtained from naïve mice by injecting 10 ml of ice-cold PBS into the peritoneal cavity and rested for 72 h at 37°C. The cells were then infected with the metacyclic forms of the parasites in a ratio of 1 : 3 and allowed for the parasites to interact for 3 h at 33°C/5%CO₂. Extracellular parasites were washed away, and the inhibitor α, β-methyleneadenosine 5-diphosphate sodium salt (APCP) was added against CD73 at a concentration of 200 μM. This treatment was left throughout the period of infection. Supernatant was collected from these groups after 48 h. The percentage of infection (a) and the amastigote number per 100 macrophages (b) were shown compared with 3 h and 48 h of incubation with or without inhibitor. Production of cytokines, (c) TNF-α, (d) IL-10, and (e) nitric oxide (NO) in treated groups is also illustrated here. Data are the from 3 independent experiments. is the statistical difference between control and treated groups using repeated measures of ANOVA followed by Newman-Keuls multiple comparison test.

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