Peritoneal Lavage and Macrophages

To verify the in vivo effects of EV Y at the site of inoculation, we extracted peritoneal lavage and macrophages from mice in the four groups described below. Uninfected mouse samples were collected 7 days after PBS or EV Y (from 10⁶ T. cruzi trypomastigotes) inoculation. Infected groups were inoculated with PBS or EV Y, and after 7 days, they were infected with 5 × 10³ T. cruzi. The peritoneal lavage and macrophages were collected at 7 dpi. After anesthesia and cervical dislocation, 2 mL of cold PBS was injected into the mouse peritoneum, the peritoneum was massaged, and the fluid was collected. Peritoneal lavage fluid was centrifuged and frozen for nitrite quantification. Macrophage immunophenotyping was performed by flow cytometry.