Effect of IL-36 and other IL-1 family members on cell proliferation and cytokine production by cultured splenocytes. Splenocytes (2 × 105 cells/well) isolated from WT (gray bars) and IL-36R−/− mice (black bars) were cultured in 96-well-plates precoated with anti-CD3 mAb (0.08 μg/mL) and incubated in the absence or presence of 100 ng/mL IL-36Ra, IL-36α, IL-36β, IL-36γ, IL-1β, IL-33, or IL-18 for 72 hours. Results are shown as fold increase compared with unstimulated cells. (A) Proliferative responses were assessed by thymidine incorporation. IFN-γ (B), IL-4 (C), and IL-17 (D) production in culture supernatants was determined by ELISA. Error bars represent the SD of the means of triplicates in 3 independent experiments. *P < .05 (Student t test), IL-36, IL-18, IL-33, or IL-1β stimulation significantly differs from unstimulated cells.
