Recombinant mouse il 25

Fluorescently-labeled antibodies against CD3 (145-2C11), CD25 (PC61), CD44 (IM7), CD16/CD32 (2.4G2), CD14 (rmC5-3), CD45R/B220 (RA3-6B2), Thy1.2 (53-2.1), ICOS (7E.17G9), Sca-1 (D7), CD69 (H1.2F3), MHC II (AMS-32.1), and Siglec-F (E50-2440) were purchased from BD Biosciences (San Jose, CA). The anti-T1/ST2 (DJ8) antibody was from MD Biosciences (St. Paul, MN). The antibodies to CD127/IL-7Rα (A7R34) and Gata3 (TWAJ) were from eBioscience (San Diego, CA). Recombinant mouse IL-7 and IL-33 were purchased from eBioscience and recombinant mouse IL-25 was from R&D Systems (Minneapolis, MN). The placebo, 17β-estradiol (E2), and combination progesterone (P4)/E2 pellets were ordered from Innovative Research of America (Sarasota, FL) and E2 powder for tissue culture was purchased from Sigma-Aldrich (St. Louis, MO).

IL-2c was prepared by combining 0.5 mg recombinant mouse IL-2 (eBioscience) with 5 mg anti-mouse-IL-2 antibody (eBioscience) and incubated at 37°C for 15 minutes. Mice were treated with isotype control, IL-2c, 0.5 mg recombinant mouse IL-33 (Invitrogen), IL-2c in combination with IL-33, 0.5 mg recombinant mouse IL-25 (R and D systems), or IL-2c in combination with IL-25 (Price et al., 2010 (link)). Mice in all treatment groups were administered agents intraperitoneally starting at 2 weeks of age. Animals were injected every other day for a total of 3 injections and were euthanized at 4 weeks of age.