INTRODUCTION The tight regulation of the cytokine network during macrophage activation is of prime importance to enable a fast and potent innate immune response against exogenous pathogens. The inflammation mediating ubiquitin-like protein HLA-F adjacent transcript number 10 (FAT10) was shown to be transcriptionally regulated by and also regulate the nuclear factor-κB (NFκB) signaling pathway. However, very little is known about the regulation of FAT10 gene expression during macrophage activation. RESULTS RNA sequencing of interferon (IFN)γ-stimulated mouse peritoneal macrophages analyzed by ingenuity pathway analysis revealed significant involvement of tumor necrosis factor receptor 1 (TNFR1) signaling in addition to IFNγ signaling. Subsequently, IFNγ robustly upregulated FAT10 expression compared to a milder induction seen with TNFα or lipopolysaccharide (LPS) stimulation. While low dose IFNγ with TNFα synergistically elevated FAT10 expression, preincubation of macrophages with IFNγ strongly augmented TNFα-induced FAT10 expression. Moreover, a short preincubation with IFNγ, which did not elevate FAT10, was sufficient to potentiate the induction of FAT10 by TNFα. A double augmentation mechanism of TNFα signaling was demonstrated, where IFNγ rapidly induced the expression of TNFα and TNFR1, which further augmented the induction of TNFα and TNFR1 expression by TNFα. Importantly, the induction of FAT10 by IFNγ in macrophages from TNFα-deficient or TNFR1-deficient mice was completely inhibited compared to macrophages from wild type (WT) mice. Finally, we show that TNFα-induced FAT10 expression is dependent on NFκB signaling. CONCLUSION IFNγ potentiates the TNFα/TNFR1 signaling pathway to induce FAT10 expression in mouse macrophages, mediated through NFκB network.

中文翻译：

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IFNγ增强TNFα/ TNFR1信号传导，从而诱导巨噬细胞中FAT10表达。

引言在巨噬细胞激活过程中，细胞因子网络的严格调节对于实现针对外源病原体的快速有效的先天免疫应答至关重要。炎症介导的泛素样蛋白HLA-F相邻转录本10号（FAT10）被转录调节，并且还调节核因子-κB（NFκB）信号传导途径。但是，关于巨噬细胞活化过程中FAT10基因表达的调控知之甚少。结果通过独创性途径分析分析了干扰素（IFN）γ刺激的小鼠腹膜巨噬细胞的RNA测序显示，除了IFNγ信号传导外，肿瘤坏死因子受体1（TNFR1）信号传导也大量参与。随后，与TNFα或脂多糖（LPS）刺激相比，IFNγ强烈上调FAT10表达。低剂量的IFNγ和TNFα协同增加FAT10的表达，而巨噬细胞与IFNγ的预温育则大大增强了TNFα诱导的FAT10的表达。而且，与IFNγ的短暂预温育并没有升高FAT10，足以增强TNFα对FAT10的诱导。证实了TNFα信号转导的双重增强机制，其中IFNγ快速诱导TNFα和TNFR1的表达，这进一步增强了TNFα对TNFα和TNFR1表达的诱导。重要的是，与野生型（WT）小鼠的巨噬细胞相比，TNFα缺陷或TNFR1缺乏的小鼠巨噬细胞中IFNγ对FAT10的诱导被完全抑制。最后，我们表明，TNFα诱导的FAT10表达依赖于NFκB信号传导。结论IFNγ增强了TNFα/ TNFR1信号通路，以诱导NFκB网络介导的小鼠巨噬细胞FAT10表达。