Fig. 3: In vitro skewing with IL-12 mediates ILC2 conversion resulting in the generation of a population of ILC1-like ILC2s. | Nature Communications

Fig. 3: In vitro skewing with IL-12 mediates ILC2 conversion resulting in the generation of a population of ILC1-like ILC2s.

From: Type II innate lymphoid cell plasticity contributes to impaired reconstitution after allogeneic hematopoietic stem cell transplantation

Fig. 3

a Schematic depicting ILC2s elicitation and isolation. b, c Representative flow cytometry plots of intracellular cytokine (b) and transcription factor expression (c) from cells expanded in vitro as described in 3 A. Error bars in (c) represent SEM. d ILC gene expression profiles based on mRNA abundance. e Differential sites of CA (two-sided Wald Test DESeq2), p adj < 0.05) between ILC2 and pcILC2s. The heatmap displays a z-score normalized ATAC signal. R1 and R2 indicate replicate numbers. f Representative tracks of normalized ATAC signal. Tick marks indicate differential sites of CA (two-sided Wald Test (DESeq2), p adj < 0.05). kb = kilobase. g UMAP of integrated RNA abundance signal from 7918 nuclei. ILC2s cultured with IL-7 and IL-33 (blue), ILC2s grown with proinflammatory cytokines, pcILC2 (orange). h Average chromatin accessibility at murine ILC2 marker genes8.The Red dot shows mean value, ILC2-pcILC2: p < 2.2 × 10−16, ILC2-ILC1-like: p = 7.46 x 10−5. i Average per cell expression of genes associated with ILC1, ILC2, shared between ILC1/2, and ILC1/38. The analysis includes 4,758 ILC2 and 5641 pcILC2 nuclei. ***p < 2.2 × 10−16. j Candidate regulators of pcILC2 cells. k UMAP of integrated ILC2 and pcILC2s annotated with normalized gene expression. l UMAP of weighted nearest neighbor integration of single nucleus RNA and ATAC from in vitro ILC2s, pcILC2s, and post-transplant cells. The black arrow highlights ILC1-like cells. m, n Lethally irradiated B6D2 mice received T cell depleted bone marrow (BM, BM only), BM plus total splenic T cells (BM + T cells), BM plus T cells with activated ILC2s (BM, T cells + ILC2) or pcILC2s (BM, T cells + pcILC2). Mean ± SEM, n = 7–16. n Kaplan-Meier survival curve after allo-HSCT. Representative of 2 independent experiments with n = 6–10 mice. Log-rank (Mantel-Cox) test, ***P < 0.01. Boxplots (h, i), are centered at median. Box bounds represent the interquartile range (IQR), whiskers represent the min/max values, and outliers are not shown.

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