Intrathymic injections were performed in 8-wk-old C57BL6 CD45.2+ or CD45.1+ WT mice. Briefly, mice were anesthetized by i.p. injection of ketamine (0,05mg/g) and xylazine (0,01mg/g) (both from Richmond Vet Pharma) in saline. An incision was opened to expose the thymus, and 10 μl of eFluor 670 (eF670) dye (0,5mM, BD Biosciences) or 10 × 106 thymocytes from OT-I mice (97–99% Vβ5+) resuspended in 20 μl PBS, were injected into CD45.1+ or CD45.2+ B6 recipient mice. The wound was closed with instant adhesive, and the mice were placed in a warm environment until they recovered. Mice were analyzed 6 days after eF670 injection or 48h after cell injections.
For IL-15 in vivo neutralization, IL-4 KO T. cruzi-infected (Tulahuen) mice were i.p. injected with 25μg of an anti-IL15 (eBioscience) resuspended in 200μl sterile 0.9% sodium chloride solution on days 10 and 12 post-infection (pi). Mice were euthanized on day 14 pi.
For IL-15 in vivo neutralization, IL-4 KO T. cruzi-infected (Tulahuen) mice were i.p. injected with 25μg of an anti-IL15 (eBioscience) resuspended in 200μl sterile 0.9% sodium chloride solution on days 10 and 12 post-infection (pi). Mice were euthanized on day 14 pi.