Kmc4021

Mice were euthanized with 100% isoflurane following the IACUC protocol #10365. After euthanasia, the head was removed from the mouse body, and the lower mandible was discarded from the snout. Nasal wash was performed by pushing 500 μL of PBS using a plastic tube attached to a 26-gauge needle through the nasopharynx into a 1.5 mL Eppendorf tube. Nasal washes were repeated 3 times. Samples were centrifuged at 3,900 rpm for 5 min at 4 °C; the supernatant was aliquoted in 100 µL aliquots and stored at −80 °C. Concentration of IFN-I in nasal washes was determined by ELISA (42120 and 42400; PBL Assay Science) as previously described. Concentration of IFN-λ in nasal washes was determined by ELISA (DY1789B; R&D Systems) as previously described. Concentration of IFN-γ in nasal washes was determined by ELISA (KMC4021; Invitrogen) according to the manufacturer’s instructions. Concentration of IL-1α in nasal washes was determined by ELISA (BMS611; Invitrogen) according to the manufacturer’s instructions.

Blood urea nitrogen (BUN) and serum creatinine were measured using specific assay kits (#K002-H1, Arbor Assays, Ann Arbor, MI, USA). TNF-α (#BMS607-3), IL-6 (#KHC-0061), and IFN-γ (#KMC4021, Invitrogen, Carlsbad, CA, USA) were measured using specific ELISA kits following the manufacturer′s instructions.

Serum levels of triglyceride (#10010303, Cayman Chemical) and total cholesterol, HDL, and LDL/VLDL (#ab65390, Abcam) were analyzed using colorimetric kits according to the manufacturer’s protocol. Serum levels of cytokines such as IL-6 (#431307, BioLegend), IFNγ (#KMC4021, Invitrogen), and TNFα (#BMS607-3, Invitrogen) were measured in male acute and chronic infected mice using ELISA kits following the manufacturer’s protocol.