Nil 27p28ab

For the allergic CHS model, mice were sensitized via topical application of 0.5% (v/v) 1-Fluoro-2,4-dinitrobenzene (DNFB) (Sigma-Aldrich) in 4:1 acetone/olive oil on their shaved back (50 µL) and were challenged 4 or 5 days later with 0.2% DNFB or vehicle control (5 µL on the dorsal and 5 µL on the ventral side of the ear). Ear thickness was measured using an engineer’s micrometer (Mitutoyo, Kawasaki, Kanagawa, Japan). The mice received neutralizing IL-27 antibody (nIL-27p28AB) (R&D Systems, Minneapolis, MN), IL-15 complex (cpx), or their respective IgG control (Ctrl). To make approximately 1 µg of the IL-15 cpx, 1 µg of IL-15 (PeproTech, Rocky Hill, NJ) and 4.5 µg of IL-15 Rα (R&D Systems) were incubated for 30 minutes at 37°C (34 (link)). Each mouse received 1.2 µg of IL-15 of the cpx.

Mice were sensitized via topical application of 0.1% (v/v) DNFB (Sigma-Aldrich) in 4:1 acetone/olive oil on their shaved back (50 µL), 5 µL on the dorsal, and 5 µL on the ventral side of the ear. Then, the mice were challenged for a total of 3 times to generate stable T cell clusters. A single dose (12 µg) of the nIL-27p28AB (R&D Systems), anti-CD122 (66 µg, BioXcell, Lebanon, NH), or goat IgG (12 µg, R&D Systems), rat IgG (66 µg, BioXcell) was injected intradermally (i.d.) on mice ears. The ears were collected for gene and flow analysis at various time points after the administration. The mouse back was injected with nIL-27p28AB or goat IgG (6 µg, daily i.d.) for a total of 3 times, and the back skin was harvested after 6 hours for immunofluorescence analysis.