Fig. 5: PD-1 controls aILC2 proliferation through metabolic regulation.

a–f ILC2s were sorted from WT and PD-1 KO mice after three intranasal challenges with 0.5 μg of rm-IL-33. Sorted cells were incubated with rm-IL-2 (10 ng mL⁻¹) and rm-IL-7 (10 ng mL⁻¹) for 24 h. a Heat map representation of differentially regulated transcription factors in FACS-sorted aILC2s from WT and PD-1 KO mice lungs. GSA algorithm was used to test for differential expression of genes (p-value < 0.05, n = 3). b–f Sorted aILC2s were cultured in vitro for 24 h with 2-DG (0.5 mM) or CYL (20 mM) in some conditions. b Representative flow cytometry plots of GATA-3 and c intranuclear Ki67 with e, f the respective corresponding quantification presented as MFI in live WT and PD-1 KO aILC2s. g–k WT and PD-1 KO mice were intranasally challenged for 3 consecutive days with 0.5 µg rm-IL-33. On day 2 and 3, mice were intraperitoneally (i.p.) injected with 2-DG (500 mg kg⁻¹). On day 4, mice were euthanized. h Representative flow cytometry plots of ILC2s gated as CD127⁺ ST-2⁺ from CD45⁺ lineage⁻ lung cells and i the corresponding quantification presented as the absolute number of ILC2s. j Quantification of GATA-3 and k intranuclear Ki67 presented as MFI in WT and PD-1 KO aILC2s. Data are representative of at least two independent experiments and are presented as means ± SEM (n = 4; two-tailed Student’s t-test or one-way ANOVA). Mouse image provided with permission from Servier Medical Art.