Trained Immunity of IL-12-, IL-15-, and IL-18-Induced CD₃+CD₅₆+ NKT-Like Cells

Figure 5

CDK4/6 inhibitors are involved in the formation of cytokine-induced trained CD3+CD56+ NKT-like cells. (a) Overview of experimental design. CD3+CD56+NKT-like cells were preactivated with rhIL-12 (10 ng/mL), IL-15 (1 ng/ml), IL-18 (50 ng/ml) ± CDK4/6 inhibitor (10um/L), or control (1 ng/ml of IL-15 alone) for 16 hours. Flow cytometry analysis was performed at the indicated time point after preactivation. (b) The gating strategy for CD3+CD56+NKT-like cells and percentages of IFN-γ+ cell populations; overlaid histograms show repressed expression of IFN-γ; percentages of IFN-γ+ cell populations declined by preactivated with the CDK4/6 inhibitor/DNA-demethylating agent/Myc inhibitor/EZH2 inhibitor; n = 8. (c) Western blot analysis of the expression of CDK4, CDK6, cyclin D1, p-Rb, and E2F-1. , , (error bars, mean ± SEM). (d) The formation of cytokine-trained CD3+CD56+ NKT-like cells，cells stimulated by IL-12/15/18 via the CDK4/6 signaling pathway induce active proliferation and differentiate into trained CD3+CD56+ NKT-like cells with enhanced function molecules expression (IFN-γ, TNF-α, GzmB, and Ki67).

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