IL-2/α-IL-2 complexes (IL-2c) were prepared as follows. About 1 ug recombinant mouse IL-2 (Peprotech) was mixed with 5 ug α-IL-2 (Bioxcell, clone JES6-1) and incubated at 37 °C for 30 min. Mice were injected intraperitoneally (i.p.) with IL-2c for 3 consecutive days and sacrificed for analysis 48 h later.
In the IL-2c treatment experiment, for blockade of cell death in vivo, 10 mg/kg Z-VAD-FMK (Apexbio Technology) and 5 mg/kg Necrostatin-1 (Abmole) or DMSO in PBS were injected i.p. daily for 5 days. In the IL-2c treatment experiment, for blockade of FASL in vivo, 400 ug IgG (Bioxcell) or α-FASL (Bioxcell) in PBS was injected i.p. on day −1, 1, and 3, and mice were sacrificed for analysis on day 5.
For IL-33 treatment in vivo, mice were i.p. injected with 500 ng recombinant murine IL-33 (BioLegend) or PBS for 4 consecutive days and sacrificed for analysis on day 5.
In the IL-2c treatment experiment, for blockade of cell death in vivo, 10 mg/kg Z-VAD-FMK (Apexbio Technology) and 5 mg/kg Necrostatin-1 (Abmole) or DMSO in PBS were injected i.p. daily for 5 days. In the IL-2c treatment experiment, for blockade of FASL in vivo, 400 ug IgG (Bioxcell) or α-FASL (Bioxcell) in PBS was injected i.p. on day −1, 1, and 3, and mice were sacrificed for analysis on day 5.
For IL-33 treatment in vivo, mice were i.p. injected with 500 ng recombinant murine IL-33 (BioLegend) or PBS for 4 consecutive days and sacrificed for analysis on day 5.