Figure 2.

Differentiation, proliferation, cytokine secretion and in vivo activity of DN ILCs. A-E: FACS-sorted DN (lin-CRTH2-IL7Rα-) cells from PBMCs obtained from an asthmatic patient (A) were cultured with IL2/IL25 for 7 days and their expression of CRTH2 and IL7Rα was analyzed (B). The newly emerged CRTH2+ (C), IL7Rα+ (E) and the remaining DN cells (D) were gated for expression of IL13. F: Results of experiments from 3 different asthmatic donors. G: FACS-sorted CRTH2+, IL7Rα+ and DN cells were cultured with IL2/IL25 for 7 days and then analyzed for expression ki67 by flow cytometry (N=3). (H) CD45+Lin- CRTH2+, IL7Ra+ and DN ILCs from the human lung were cultured with IL2/IL25 for 5 days and then supernatant was assayed for IL5 by ELISA. Culture supernatant from IL2/IL25-treated B cells (negatively selected from PBMCs) was used a control. Each symbol represents a single lung donor. (I) Human lung-derived ILC2 subpopulations were cultured with IL2 overnight and adoptively transferred (10⁵ cells/mouse) to Rag2^−/−:γc^−/− mice. Lung-derived CD45+CD3+CD4 T cells were injected (10⁶ cells/mouse) as a control. One-day latter the mice were challenged with the Alternaria allergen for 3 consecutive days before methacholine challenge. The increase in lung resistance was statistically significant (*=P<0.05, t test) for all ILC2 populations as compared to the control (CONT).