Fig. 5: Acute insult activates thymic NK cells.
From: Damage-induced IL-18 stimulates thymic NK cells limiting endogenous tissue regeneration
a, Normalized gene expression in NK/ILC1 or NKT cells of the cytotoxicity factors Ifng, Prf1, Gzma and Gzmb, as well as the activation markers Nkg7, Klrd1, Klrk1, Ncr1, Klrc2 and Klra4, on days 0, 1, 4 and 7 after SL-TBI, taken from the scRNAseq dataset described in Fig. 2a. b, Concatenated flow cytometry plots and corresponding geometric mean fluorescence intensity (gMFI) of CD45+NK1.1+TCRβ− NK cell expression of Ifng-GFP (day 0, n = 4; day 3, n = 5), perforin (day 0, n = 6; day 3, n = 9) and granzyme B (GZMB; day 0, n = 4; day 3, n = 5) on days 0 and 3 following SL-TBI in female 1- to 2-month-old C57BL/6 WT or Ifng-reporter mice. c, Amount of thymic IFNγ (day 0, n = 8; day 3, n = 3), perforin (n = 8 per group) and granzyme B (n = 8 per group) measured by ELISA in female 1- to 2-month-old C57BL/6 mice on days 0 and 3 after SL-TBI. d, Thymuses were collected from female 1- to 2-month-old C57BL/6 mice on day 3 after SL-TBI. Concatenated flow cytometry plots gated on all CD45+ Ifng-GFP (left), CD45+perforin+ (middle) and CD45+GZMB+ (right) cells, as well as the total thymus cellularity of Ifng-GFP (left; day 0, n = 3; day 3, n = 4), perforin+ (middle; day 0, n = 3; day 3, n = 7) and GZMB+ (right; day 0, n = 3; day 3, n = 7) CD45+NK1.1+TCRβ−CD49b+ NK cells, CD45+NK1.1+TCRβ−CD49a+ ILC1s, CD45+NK1.1+TCRβ+CD49b− NKT cells and CD45+TCRβ+NK1.1− T cells, are shown. e, Gene expression heat map of thymic NK/ILC1 and NKT cells for Gzma, Gzmb, Prf1, Ifng, Nkg7, Klrk1, Ncr1, Klrd1 and Il18r1 at 1, 4 and 7 days following SL-TBI. Each column represents a cell, with the cells ordered based on the expression of Il18rap (in ascending order from left to right). The time after TBI is indicated by the colors at the bottom. f, NK1.1+IL-18R+TCRβ−CD49b+ NK cells from female 1- to 2-month-old C57BL/6 mice were purified using FACS at baseline (d0) or 2 days after SL-TBI (d2) and cocultured with CellTrace-labeled RMA-S target cells at a 2:1 effector-to-target ratio. RMA-S target cell Annexin V expression was measured 5 h after coculture, and cell death was assessed (n = 4 biological replicates per group, representative of three independent experiments). Dashed lines represent RMA-S alone (bottom) or the positive control (top). Graphs represent mean ± s.e.m.; each dot represents an individual biological replicate. Statistics were generated for b–d and f using unpaired two-tailed t tests.
