Anti fcr blocking reagent

Lung cells were isolated from Alt-treated Rag1 KO mice after the last sensitizing exposure (Fig. 1 A) according to the protocol described above. Mouse lung single-cell suspension was blocked with an anti-FcR blocking reagent (#130-092-575; Miltenyi Biotec) before staining. ILCs were stained with FITC-labeled anti-CD45.2 (clone 104), PerCP-Cy5.5–conjugated anti-CD25 (clone PC61.5; eBioscience), Pacific Blue anti-mouse NK-1.1 antibody (#108722; BioLegend), or PE-Cy7–labeled anti-NK1.1 (#108714; BioLegend), anti-ST2 (#FAB10041P-100UG; clone #245707; R&D), and APC-Cy7–labeled anti-ICOS (clone C398.4A) antibodies. Live CD45.2, CD25, and ICOS⁺ST2⁺ and ICOS⁻ST2⁻ cells were sorted on a BD FACSAria Fusion and delivered i.v. (50,000 cells/mice) to Rag2^−/−γc^−/− mice 24 h after sorting. Intranasal Alt (2.5 µg in 20-µl dose in PBS) was administered on three consecutive days after i.v. transfer of ICOS⁺ST2⁺ and ICOS⁻ST2⁻ cells. The recipient mice were studied on day 6 for measurement of airway hyperreactivity, inflammation, and type 2 cytokine production.