Quantification of Allergic Immune Markers

Cytokine, chemokine and antibody concentrations were measured via ELISA. Plasma collected at the time of sacrifice was assayed for OVA-specific IgE and IgG₁ similarly to previously described.^{30 (link)} Briefly, plates were coated with 100 μg/ml OVA, and blocked with 1% BSA, prior to incubation with diluted plasma. Bound antibody was detected with biotinylated rat anti-IgG₁ (clone A85–1, BD Pharmigen) or rat anti-IgE (clone R35–118, BD Pharmigen), followed by addition of streptavidin-HRP (BD Pharmigen) and supersensitive TMB (Sigma-Aldrich) as a substrate. Lung homogentates were prepared from half of the left lung as previously described^{27 (link)} in T-PER buffer (ThermoFisher Scientific, Waltham, MA, USA). ELISAs for IL-5 (eBioscience), eotaxin (R&D Systems, Minneapolis, MN, USA), and RANTES (R&D Systems) were performed according to the manufacturers’ instructions.