Figure 3.

Human complement activation by N. annulifera venom (NaV) results in the generation of chemokines and lipid mediators. Human whole-blood samples were incubated for 5 minutes, at room temperature, with Cp40 (C3 cleavage inhibitor), SB290157 (C3aR antagonist), PMX205 (C5aR1 antagonist), P32 (C5aR2 agonist) or 1,10 Phenanthroline (metalloproteinase inhibitor) inhibitors, or their respective vehicles. Then, all these samples were exposed to NaV or sterile saline during 30, 60 and 120 minutes, and complement activation products generation (A–C), lipid mediators release (D–F) and chemokines upregulation (G–I) scrutinized by CBA or ELISA. Data are means ± SEM of six independent experiments with different whole-blood donors. ***p ≤ 0.001 (two-tailed two-way ANOVA, followed by Bonferroni post-test). ^### indicates a significant difference between N. annulifera venom + Vehicle and N. annulifera venom + Inhibitors. (E, H) ^## means the comparison between samples exposed to the NaV + vehicle and NaV+ complement inhibitors in which statistical differences are p ≤ 0.01. ns = non-significant.