Ifn λ3

For depletion of CCR2⁺ cells, CCR2-DTR mice and control CCR2-DTR negative littermates received 250 ng of diphtheria toxin i.p. one day prior to infection and every other day thereafter in order to maintain depletion. Mice were injected i.p. with 1.0 µg of either IFNα2 (Novoprotein), IFNλ3 (PBL), or both on D0 and D+1 for ROS analysis 48 hours post-infection. For survival, mice were injected i.p. with 1.0 µg of either IFNα2, IFNλ3, or 500 ng of both for a total of 1.0 µg on D0 and every other day. Mice were euthanized as they developed severe symptoms of IA, and survival was terminated at D+14.
For CCR2⁺monocyte transfer experiments, CD45.1⁺CCR2-GFP⁺ monocytes were sorted from mice that were infected with 4 × 10⁷Af conidia for 3 hours (peak of IFN-α transcription). CD45.1⁺CCR2-GFP⁺ monocytes (~850,000) were transferred via retro-orbital injection into CD45.2⁺CCR2-DTR mice that have been infected with 4 × 10⁷Af conidia for 3 hours. CD45.2⁺CCR2-DTR mice were treated with DT on D-1, D0, and D+1. Congenic markers were used to track transferred cells in recipients by flow cytometry. ROS and fungal burden was assessed 48 hours after infection. Monocytes were sorted as (Live CD45.1⁺CD11b⁺NK1.1⁻CCR2-GFP⁺Ly6C⁺)