Figure 5

Importance of ILC2s for large-size chitin-induced airway eosinophilia. Mice were intranasally treated one time with 100 μg of chitin (large size: 70–100 μm) suspended in saline or with saline alone. Twenty-four hours later, BAL fluids and lungs were collected. (a) Numbers of eosinophils and neutrophils in the BAL fluids from C57BL/6N-wild-type (n = 9) and Kit^W-sh/W-sh (n = 4) mice. (b) Numbers of eosinophils and neutrophils in the BAL fluids from C57BL/6J-wild-type (n = 22) and Rag2^−/− (n = 19) mice. (c) Numbers of eosinophils and neutrophils in the BAL fluids from C57BL/6J-wild-type (n = 10) and Rag2^−/−Il2rg^−/− (n = 8) mice. (d) Proportion of IL-5-venus-positive cells in CD45⁺ CD25⁺ lineage marker-negative ST2⁺ cells (ILC2s) in the lungs from naïve wild-type and saline- and chitin-treated Il5^venus/+ mice by FACS. (e) Number of ILC2s in the lungs from saline- and chitin-treated Il5^venus/+ mice (n = 5). (f) Proportion of IL-5-venus-positive cells in CD45⁺ CD3⁺ CD4⁺ T cells in the lungs from naïve wild-type and saline- and chitin-treated Il5^venus/+ mice by FACS. (g) Number of Th2 cells in the lungs from saline- and chitin-treated Il5^venus/+ mice (n = 5). (a–c,e,g) Data from 2 independent experiments were pooled and shown as the mean + SE *P < 0.05 vs. wild-type mice. (d,f) Data show a representative result from the lungs of 5 mice. (d,f) Cells were analyzed on a FACS Aria II Cell Sorter (BD Biosciences) with BD FACSDiva v6.1.3 software (BD Biosciences) and FlowJo v10.5.3 software (BD Biosciences; https://www.flowjo.com/solutions/flowjo).