Fig. 5: lnc13 restrains IL-15-induced differentiation of CD8+ NK-like lymphokine-activated killer cells.
From: The long noncoding RNA lnc13 restrains inflammatory responses to maintain oral tolerance to gluten
a, Relative expression of lnc13 in murine CD4+ T cells, CD8+ T cells and B cells, measured by RT–qPCR. lnc13 is most highly expressed in CD8+ T cells (n = 4; P = 0.00700709 comparing CD4+ versus CD8+ T cells, P = 0.00795608 comparing CD8+ T cells and B cells). b, RT–qPCR quantification of lnc13 and Dcp2 expression in CD8α+ IELs cultured for 48 h with medium alone, soluble IL-15 (sIL-15) or IL-15 superagonist (IL-15SA; IL-15 pre-complexed with IL-15Rα). IL-15 stimulation leads to downregulation of lnc13 and upregulation of its degrading enzyme Dcp2. lnc13 and Dcp2 expression, by RT–qPCR, was first normalized to Rpl32. ΔΔCt was calculated relative to average of unstimulated samples (for lnc13) or IL-15SA stimulated samples (for Dcp2). n = 5 mice per experiment. For comparisons of lnc13 expression medium versus IL-15 and medium versus IL15ra, as well as Dcp2 medium versus IL-15 and medium versus Il15rap, P = 4.70678 × 10−6, 2.74899 × 10−6, 2.47 × 10−5 and 4.7847 × 10−7, respectively. c, Schematic model proposing that IL-15 signaling activates STAT5 to promote Dcp2 expression, leading to degradation of lnc13. d, Time-course of NKG2D expression in WT CD8a+ IELs cultured with or without IL-15SA. NKG2D induction becomes robust by day 5, marking NK-like differentiation. e, Purified CD8a+ T IELs were stimulated with IL-15SA for 5 days. CD8+ IELs from lnc13−/− mice exhibit enhanced cytotoxicity and NK marker expression. Representative flow plots (left) and quantification (right) showing: NKG2D+GZMB+ double-positive live cells. n = 4 mice across three independent experiments. P = 0.00028843. Samples from lnc13−/− mice are represented by red dots and those from lnc13+/+ mice are represented by blue dots. f, After 5 days of IL-15SA stimulation, representative histogram of GZMB and NKG2D of WT-DQ8 (blue) and lnc13−/−-DQ8 (red) is shown. Dashed line represents isotype control. Error bars represent mean ± s.e.m. Unpaired, two-tailed Student’s t-test was used for comparisons. **P < 0.01, ***P < 0.001, ****P < 0.0001.
