Panel A shows UMAP of distinct ILC2 clusters in infected control and Blimp-1–deficient mice. The overall distribution of clusters is similar, but subtle shifts suggest altered cellular states in cKO mice. Panel B shows cluster frequency analysis illustrating changes in the proportion of specific ILC2 subsets between control and cKO mice, indicating that Blimp-1 loss affects population composition during infection. Panel C shows heatmap of Cluster-specific marker gene expression patterns highlight functional heterogeneity among ILC2 subsets. Distinct gene signatures define activated, proliferative, and effector-like clusters. Panel D shows GO analysis indicates that Blimp-1–deficient ILC2s are enriched for pathways related to cell cycle and proliferation, while control cells show enrichment in immune activation and differentiation pathways. Panel E shows flow cytometry plots confirms increased Ki67 expression in cKO ILC2s, demonstrating enhanced proliferation in both mLN and lung following infection. Panel F shows UMAP of reduced expression of effector cytokines (Il5, Il13) in cKO ILC2s, while expression of receptors such as Il1rl1 and Il2r a varies across clusters. Panel G shows violin plots further illustrate diminished Il5 and Il13 expression across multiple clusters in Blimp-1–deficient ILC2s compared with controls. Panel H shows quantitative analysis of lung ILC2s confirms significantly reduced Il5 and Il13 expression in cKO mice, indicating impaired effector function despite increased proliferation.
Blimp-1–deficient ILC2s proliferate but exhibit reduced effector functions. (A) UMAP of scRNA-seq of mLN ILC2s from infected Ctrl (Prdm1fl/fl) and cKO (Nmur1CrePrdm1fl/fl) mice at day 7 of N. brasiliensis infection. Annotation of clusters on the right. (B) Frequencies of cells per cluster of samples shown in A. (C) Heatmap showing normalized count distributions of marker genes in different clusters in scRNA-seq of samples shown in A. (D) GO pathway analysis of DEGs between infected Ctrl and infected cKO mice in scRNA-seq of samples shown in A. (E) Flow cytometric plots and quantification of Ki67+ ILC2s in the mLN and lung in untreated and in infected Ctrl and infected cKO mice. Mice were infected with N. brasiliensis for 7 days. Data are representative of two independent experiments (n = 3–6 mice per group). (F) UMAPs of Il5, Il13, Il1rl1, and Il2ra expression. (G) Violin plots showing normalized count distributions of Il5 and Il13 in different clusters in scRNA-seq of samples shown in A. (H) Relative expression of Il5 and Il13 in lung ILC2s from infected Ctrl and cKO mice. Mice were analyzed 7 days after infection with N. brasiliensis (n = 4 mice per group). N. b., N. brasiliensis. Mean ± SD; one-way ANOVA. Student’s t test; *P < 0.05; **P < 0.01; ****P < 0.0001. UMAP, uniform manifold approximation and projection.