Mice and T. cruzi infection

BALB/c mice obtained from the Comisión Nacional de Energía Atómica (CNEA; Buenos Aires, Argentina) were inbred and housed according to institutional guidelines. BALB/c mice, when 6–8 weeks old, were intraperitoneally infected with 1x10⁶ blood-derived T. cruzi trypomastigote forms from Tulahuén strain, which was maintained through intraperitoneal inoculation every 11 days [32].

Female BALB/c mice were infected intraperitoneally with 500 blood-derived T. cruzi trypomastigote forms diluted in saline solution. After different days post infection (p.i.), these mice were sacrificed by CO₂ asphyxiation and spleens were extracted. Non-infected animals were processed in parallel. Trypomastigotes of the Tulahuen and Y strains were obtained from the extracellular medium of infected monolayers of Vero or NIH3T3 cells, respectively, and were collected by centrifugation at 4400 rpm for 10 min and resuspended in RPMI medium containing 10% FCS. Parasites were counted using a Neubauer chamber and used for in vitro infection experiments as described below.