Fig. 8: Adenosine promotes intracellular cAMP accumulation and suppresses ILC2 through NF-κB signal pathway.

a Lung ILC2 were cultured in the presence of IL-2, IL-7 and IL-33 and treated with DMSO or NECA (1 µM) for 24 h, cells were collected for transcriptional profiling analysis by SMART-seq. Heatmap visualization of the distribution of cells from the four different samples and different expressed genes related to cAMP signal. Lung ILC2 were culture in the presence of IL-2, IL-7 and IL-33 with NECA (1 µM) or DMSO treatment for 3 days, the cAMP concentration in supernatants (b) and ILC2 lysate (c) were measured by ELISA. d–f Lung ILC2 were culture in the presence of IL-2, IL-7 and IL-33 with adenylate cyclase inhibitor SQ22536 (10 µM) for 3 days, Flow cytometric analysis of intracellular IL-5⁺IL-13⁺ ILC2 (d) and Ki67⁺ ILC2 (e). The amount of IL-5 and IL-13 in supernatants were measured by ELISA (f). g Heatmap of different expressed genes related to NF-κB signal pathway. h Lung ILC2 were cultured in the presence of IL-2, IL-7 and IL-33 for 6 h and treated with DMSO, NECA (1 µM)+DMSO, NECA + SCH58261 (1 µM) or NECA + NF-κB agonist Betulinic acid (BA,10 µM)/Prostratin(10 µM), Flow cytometric analysis of the p-P65 MFI in ILC2 with indicated treatment. The amount of IL-5 and IL-13 in supernatants were measured by ELISA (i, j). k WT mice were challenged with papain i.n for 5 consecutive days and simultaneously treated with NECA or DMSO every day intraperitoneally, p-P65 MFI in lung ILC2 were analyzed. Data are representative of two independent experiments. Error bars show mean ± SEM; *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired Student’s t test (b, c) or one-way ANOVA with Bonferroni post-test (d–j).