Figure 7.

Interaction between complement system and peptidorhamnomannans (PRMs) of Sporothrix species. (A) Sporothrix yeasts were opsonized with C3b and labeled with mouse anti-C3b mAb and mouse TRITC-IgG (Ab). PRMs (50 µg/ml in 50 mM carbonate buffer, pH 9.6) were coated on 96-well plate: their (B, C) C3 activation capacities were determined using GVB+ or GVB- (with EGTA) media using whole/complement factor-depleted human sera. (D) Interaction with CD11b was determined by sequential addition of CD11b, mouse anti-CD11b mAb, peroxidase-conjugated mouse IgG, and chromogenic substrate for peroxidase (tetramethyl benzidine); colors developed were optically read at 450 nm. For each condition, two independent experiments, in triplicate, were performed. Statistical analysis was performed by ANOVA with Tukey’s multiple comparison posttest (****p < 0.0001).