Figure 1. . IFNγ is sufficient for T. gondii-induced inflammasome activation, IL-1β release and lytic host cell death in BM-MoDCs.

B6 bone marrow-derived monocytes/dendritic cells (BM-MoDCs) were pretreated for 20 hours with 70ng/mL IFNγ with or without 100ng/mL of Pam3CSK4 (Pam) for the final 3 hours before infection or incubated in media alone. Cells were infected with Me49 expressing GFP and luciferase (Tg) or treated with an equivalent volume of supernatant during parasite preparation (sup) to control for released danger-associated molecular patterns (DAMPs). At 14 hours post-infection (hpi), supernatant was harvested to assess cell death by lactate dehydrogenase release (LDH) or IL-1α and IL-1β protein levels by ELISA (A). Controls were pre-treated with 500ng/mL LPS for 3 hours, with IFNγ or IFNγ and Pam similarly above, followed by 1 hour stimulation with 5μM Nigericin (Nig). B, Cell pellets were collected or supernatants were TCA-precipitated to measure protein levels by Western blot. # indicates non-specific bands. Representative of 3 independent experiments. Error bars represent Mean±SEM by one-way ANOVA with Tukey post hoc analysis and nigericin-treated groups were analyzed separately. *, p≤0.05; **, p≤0.01; ***, p≤0.001; ****, p≤0.0001.