Figure 5.

(A) Cytokine released by human monocyte-derived macrophages (hMDMs) blocked for complement receptor-3 (CR3) and challenged with Sporothrix species. Sporothrix yeasts were made to interact with hMDMs blocked for CR3 (with monoclonal anti-CD11b antibodies) or unblocked (infected control) in a culture medium supplemented with whole human serum (wHS) for 18 h at 37°C and 5% CO₂. Non-infected controls were hMDMs blocked with irrelevant antibodies and treated with culture medium supplemented with wHS. The culture supernatants were collected and analyzed for tumor necrosis factor (TNF)-α and interleukin (IL)-1β. Results indicate the mean ± SEM of at least two independent experiments in duplicate or triplicate (n = 4 or 6). ANOVA with Tukey’s multiple comparison posttest was performed for the statistical analysis (***p < 0.001, **p < 0.001). (B) Uptake of Sporothrix yeasts by hMDMs after CR3 blockage. hMDMs were incubated with monoclonal anti-CD11b antibodies for 1 h and challenged with FITC-labeled yeasts of Sporothrix schenckii or Sporothrix brasiliensis. After 4 h of interaction in the presence of medium containing wHS, hMDMs were analyzed by FACS to determine the change in mean binding index (MBI) compared to control (=CD11b unblocked hMDMs interacted with FITC-labeled Sporothrix yeasts). Results are the mean ± SD of an experiment performed in duplicate.