Figure 6.

Phagocytosis of Sporothrix schenckii wild-type, control, and ROT2-silenced strains by human monocyte-derived macrophages. In (A), yeast-like cells were labeled with Acridine Orange, coincubated with human monocyte-derived macrophages at a macrophage–fungus ratio of 1:6, for 2 h at 37 °C and 5% (v/v) CO₂. Human cells were gated by FACS, and 50,000 events, defined as a human cell interacting with at least one fluorescent fungal cell, were counted per sample. Control, macrophages interacting with no yeast-like cells. * p < 0.05 when compared to WT, HSS31, or HSS32 strains. ^† p < 0.05 when compared with HSS33, HSS34, or HSSS35 strains. In (B), experiments described in panel A were performed, but macrophages were previously preincubated with 10 μg mL⁻¹ of any of the following antibodies: anti-mannose receptor, anti-CR3, anti-TLR2, or anti-TLR4. Alternatively, the human cells were preincubated with 200 μg mL⁻¹ laminarin. In all cases, the interactions were performed in the presence of 5 μg mL⁻¹ polymyxin B. No treatment refers to cells preincubated only with 5 μg mL⁻¹ polymyxin B. Results correspond to cells in the late stage of phagocytosis. For all cases, 100% corresponds to the system with no treatment, and the absolute values were similar to those shown in panel A. CR3, complement receptor 3. * p < 0.05 when compared to the no treatment condition of the same strain. For both panels, data represent means ± SD from six donors assayed by duplicate.