Fig. 5

Cytokine production by macrophages infected with L. amazonensis in vitro. Thioglycollate-elicited macrophages were harvested from the peritoneal cavity of WT or gp91^phox−/− mice 3 days after stimulation. The cells were incubated  in 24-well plates for 16 h at 37 ^oC, 5 % of CO₂ and then washed for non-adherent cells removing. The adherent cells were stimulated or not with 50 U/ml of IFN-γ plus 100 ng/ml of LPS for 24 h at 37 ^oC, 5 % of CO₂. Macrophages were infected with L. amazonensis metacyclic promastigotes (10 parasites per macrophage) for 4 h and cultures were washed and re-stimulated or not with IFN-γ plus LPS at the same concentrations used before. After 48 h of infection supernatants were collected and used to measure cytokines by ELISA for IL-6 (a), TNF (b), IL-17 (c), CXCL-1 (d), MCP-1 (e) and IL-10 (f). Data are shown as mean ± SD of one representative experiment of four, n = 5 for each experiment