In vivo Antibody Treatment

Groups of age-matched male Rora^CreRosaYFP mice were treated, using a method previously described (20), with IL-4/anti-IL-4 monoclonal antibody (mAb) complexes (IL-4c), thioglycollate (4% w/v; Sigma, Wicklow, Ireland), or thioglycollate + IL-4c i.p. IL-4c was prepared by incubating 5 μg recombinant mouse IL-4 (Peprotech, London, UK) with 25 μg anti-lL-4 mAb (clone 11B11, BioXcell, NH, USA). Briefly mice were injected with PBS or IL-4c i.p. on days 0 and 2. Mice were also treated with thioglycollate (day 0) or thioglycollate (day 0) plus IL-4c (day 0 and 2). Peritoneal cells were collected from all mice on day 4 by lavaging the peritoneal cavity with ice-cold PBS for flow cytometry.