Fig. 5. The d.c. HDM treatment induces the infiltration of IL-1β-expressing Gr-1^hi and Gr-1^int cells in the skin.

a–c Analyses of IL-1β-expressing cells in the skin. Wildtype Balb/c mouse ears were subjected to d.c. or e.c. HDM treatment and were analysed 24 h later. a Left, frequencies of eosinophils (Eos), basophils (Baso), TCR-β⁺ T, Gr-1^hi and Gr-1^int cells among alive singlets. Right, representative FACS plots. n = 4 mice. b Histogram comparison of IL-1β in cells from the d.c. HDM-treated ears. c Frequencies of IL-1β⁺ cells among alive singlets (n = 4 mice). d–g Depletion of neutrophils and monocytes/macrophages reduces IL-1β in d.c. HDM-treated ears. d Experimental protocol. Wildtype Balb/c mice were intraperitoneally (i.p.) injected with PBS, NIMP-R14 (anti-Gr-1) or anti-Ly6G antibody (Ab) at day (D) -1. Ears were d.c. HDM-treated at D0 and analysed at D1. e Representative FACS plots showing the depletion of both Gr-1^hi and Gr-1^int cells by NIMP-R14 Ab, or of Gr-1^hi cells by anti-Ly6G Ab. f Frequencies of cells among ear alive singlets (n = 4 mice). g IL-1β and TSLP protein levels in ears (n = 4, 10, 6, 4 mice). h–j Infiltration of IL-1β-expressing cells is TSLP-independent. Ears of WT or Tslp^−/− mice were treated with e.c. HDM or d.c. HDM at D0 and analysed at D1 for IL-1β (h) and for Gr-1^hi and Gr-1^int cells (i). For h, n = 6, 6, 8, 8 mice. For i, n = 6, 6, 10, 10 mice. j Recombinant TSLP or PBS were administrated on e.c. HDM-sensitized ears and IL-1β levels were measured (n = 8 mice). Graphs in a, c, f–j show mean ± SEM. a, f–i One-way ANOVA test; c, j Two-sided Student’s t test. Data are representative of 3 (a–c, e–i) or 1 (j) independent experiments with similar results. Source data are provided as a Source Data file.