(A) Migration assays of neutrophils incubated for 16 hours with media only, supernatants from uninfected HFF cells, supernatants of HFF cells infected for 4 days with T. cruzi CL-14 or CL Brener, and with purified CCL2 (500 ng/mL). (B-D) NO (B) and ROS assays (C-D) of neutrophils incubated for 16 hours with media only, supernatants from uninfected HFF cells, supernatants of HFF cells infected for 4 days with T. cruzi CL-14 or CL Brener and then challenged with 200 ng/mL LPS for one hour. IL-1β (E) and TNF-α (F) production in PMNs incubated for 24 and 96 hours with supernatants of HFF cells infected for 4 days with T. cruzi CL-14 or CL Brener. No TNF-α nor IL-1β detected in PMNs treated with media only or supernatants from uninfected HFF. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 (one-way ANOVA with Tukey’s post-test comparing the indicated treatments). • p < 0.05 and •••• p < 0.0001 (unpaired t-test comparing the indicated treatments). (C) Image is representative of three independent experiments. (A) Data from four independent experiments (mean and s.e.m.). (B, D-F) Data from three independent experiments (mean and s.e.m).