Fig. 5

IL-33 is a potent stimulator of type 2 cytokine and chemokine release from group 2 innate lymphoid cells. ILC2 and total ILC were enriched from lung tissue and stimulated with various ILC-activating cytokines. A ILC2 from untreated mice were purified using FACS sorting (as in Fig. 3) then stimulated with IL-2 in combination with IL-25, TSLP or IL-33 for 5 days [10 ng/mL, each]. IL-13 was measured in cell culture media by ELISA B Lung ILC2 isolated from Saline, RSV, OVA and OVA-RSV treated animals were stimulated with IL-2, IL-7 and IL-33 [10 ng/mL] for 4 days in culture. Cell pellets were counted at the end of 72 h to account for changes in cell divisions or proliferation across groups. IL-5, IL-13, CCL17 and CCL22 levels were assessed by ELISA. Total pg/mL detected in 50 uL of cell culture fluid, was divided by the total number of cells counted in each well at the end of 72 h to determine the pg/cell for each treatment. C Total ILC were enriched from the lungs of each subset of animals and stimulated with IL-2, IL-7 and IL-23. IL-22 and IL-17 were measured in cell culture media by ELISA. Each bar represents the mean with standard error of the mean. Cells were pooled from 2 to 3 mice per experiment. Data represent two separate experiments; N = 5–6 samples per bar. *Indicates a statistically significant difference vs. media control (P < 0.05). P-value over bar for a difference between groups determined by ANOVA