Fig. 3: IL-33 activates lymphoneogenic ILC2s.
From: IL-33-activated ILC2s induce tertiary lymphoid structures in pancreatic cancer
a, Single-cell RNA-sequencing (scRNA-seq) analysis of purified tumour and DLN ILC2s from rIL-33-treated PDAC mice. The uniform manifold approximation and projection (UMAP) representations show cells by unsupervised clusters (top left), organs (bottom left) and select genes (right). The heat map (middle) shows the top 25 genes by cluster. The heat map scale bar represents the z score. The UMAP scale bar represents gene expression. b, Gating (left) and LT expression by flow cytometry (right) and confocal imaging (bottom) in intratumoural KLRG1+ ILC2s from rIL-33-treated WT and Ltb−/− PDAC mice. FMO, fluorescence minus one; MFI, mean fluorescence intensity. c,d, Gating, and the frequency and number of intratumoural (c) and gut (d) ILC2s in vehicle-treated and rIL-33-treated PDAC (c) and DSS-colitis (d) mice. e, Gating (left), ILC2 frequency (middle) and intratumoural TLS density (right) in rIL-33 and DT-treated Nmur1icre-eGFPRosa26LSL-DTR and littermate control Rosa26LSL-DTR PDAC mice. f, Intratumoural TLS density (left), gating, frequency and the number of ILC2s, and the number of DLN immune cells (right) in rIL-33-treated Nmur1icre-eGFPLtbfl/fl and littermate control PDAC mice. g, Tumour KLRG1+ ILC2s were sort-purified from rIL-33-treated Il7rcre/+Ltbfl/fl or Il7rcre/+Ltb+/+ littermate PDAC mice and transferred to Il7rcre/+Rorafl/fl PDAC mouse recipients. The TLS density, KLRG1+ ILC2 frequency and number in recipient PDACs. Data were collected at 10 days (a), 2 weeks (b and d–f) and 5 weeks (c) after tumour implantation, and 2 weeks (g) after transfer, pooled from ≥3 independent experiments with n ≥ 3 mice per group with consistent results. n represents the number of individual tumours or organs from individual mice analysed separately. The horizontal bars show the median. In f, littermate controls include Ltbfl/+ and Nmur1icre-eGFPLtbfl/+ combined. P values were calculated using two-tailed Mann–Whitney U-test (b, e (right), f and g) and two-way ANOVA with Holm’s test (c, d and e (left)). Scale bars, 1 μm (b (left)), 5 μm (b (all others)), 20 μm (e and f (left)) and 100 μm (f (right)).
