Extended Data Fig. 3: ILC2 suppress NK cells via an indirect innate immune mechanism.

a, Il7ra^Cre/+ or Il7ra^Cre/+Rora^fl/fl mice were treated with PBS or Asp on day 0 and 1, followed by quantification of IFNγ⁺ lung NKcells on day 3 (n = 12,13,11,12). b,c, WT mice were treated as in (a), followed by purification of lung ILC2 (flow cytometry) and lung NK cells (magnetic bead, see Methods) on day 3. Lung NK were cultured alone or with ILC2 and analyzed for intracellular IFNγ and GzmB (c) (n = 8). d, WT mice were treated as in (e) with PBS or IL-33 and A2AR antagonist or DMSO followed by quantification of IFNγ⁺ lung NK cells on day 3 (n = 6,7,6). e, WT mice were treated as indicated, followed by quantification of GzmB⁺ lung NK cells (n = 3, representative gating on right). f, WT and Foxp3^DTR mice were treated with PBS or IL-33 and DTx, followed by quantification of percent IFNγ⁺ lung NK cells on day 3 (n = 3,3,4,5). g, WT and Rag2^–/– mice were treated with PBS or IL-33 on days 0 and 1, and given adoptive transfer of LL/2 cells (i.v.) on day 7. Tumor burden was assessed on day 21 by visual quantification of lung metastases (n = 10,8,8,10). h, WT, Rag2^–/– and Rag2^–/–Il7ra^Cre/+Rora^fl/fl mice were treated with PBS or IL-33 on day 0 and 1, followed by flow cytometry analysis for the indicated lung lymphoid cells on day 3 (n = 5). Bar graphs indicate mean (±S EM) and show combined data of two (c, d, and g) or three (a) independent experiments. (e, f, and h) shows a representative bar graph of two independent experiments. Statistical analyses were calculated using one-way ANOVA with ns = not significant, and **** = p ≤ 0.0001.