Figure 2. Augmented T_H2 cytokine production by ILC2s in neutrophil depleted mice administered HDM for 1 week.

(A) Balb/c mice were administered HDM or PBS intranasally (i.n.) 3 times per week for 1 week and at 24 hours prior to each HDM treatment, mice were dosed with either 100 μg of neutrophil depleting antibody, 1A8, or isotype control antibody, 2A3 intraperitoneally (i.p.). At 24 hours post the final HDM administration BALF was collected (Ŧ). (B) Concentrations of IL-4, IL-5 and IL-13 protein were assessed in and BALF by ELISA. In some experiments, CD4⁺ T cells and ILC2s (pooled from 3 mice per data point) were isolated from the airways by FACS, at 24 hours post final HDM exposure, for subsequent mRNA gene expression analysis. (C) Relative expression of Il-4, Il-5, Il-13 and Gata-3 in T cells and ILC2s derived from BAL of HDM treated mice, as determined by qPCR. At the same time point, the number of IL-13⁺ ILC2s and IL-5⁺ ILC2s (D) and geometric expression of IL-13 and IL-5 in ILC2s (E) in the BAL was assessed by flow cytometry. Figures present combined data from 2 independent experiments with 4-6 mice per group in each experiment (B, D and E), or from one experiment whereby each data point represents cells pooled from 3 independent mice (C). Results depicted as mean ± SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 using Mann–Whitney statistical test.