Sequential conditioning-stimulation reveals distinct gene- and stimulus-specific effects of Type I and II IFN on human macrophage functions
Sequential conditioning-stimulation reveals distinct gene- and stimulus-specific effects of Type I and II IFN on human macrophage functions
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DOI：
10.1038/s41598-019-40503-y
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发表时间：
2019-03-27
影响因子：
4.6
通讯作者：
Hoffmann, Alexander
中科院分区：
文献类型：
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作者：
Cheng, Quen;Behzadi, Faraz;Hoffmann, Alexander
巨噬细胞通过感知和响应病原体相关分子来协调免疫反应。这些反应通过预先用干扰素 (IFN) 等细胞因子进行调节来调节。 I 型和 II 型干扰素在许多生物学场景中具有相反的功能，但直接用 I 型或 II 型干扰素刺激的巨噬细胞会激活高度重叠的基因表达程序。我们假设序贯条件刺激方法将更特异性地揭示 I 型和 II 型 IFN 对人类巨噬细胞的不同影响。首先用 IFN 进行调节，然后用 Toll 样受体配体和细胞因子进行刺激，然后进行全基因组 RNA-seq 分析，我们鉴定了 713 个基因，其表达不受单独的 IFN 影响，但在调节后对刺激的反应显示出增强或减弱。例如，对细胞因子 TNF 的反应受到 II 型 IFN 调节的限制，但受到 I 型 IFN 调节的增强。我们观察到干扰素的作用并不统一是促炎或抗炎，而是高度基因特异性和刺激特异性。通过评估关键信号转导器的表达水平并通过 ATAC-seq 表征染色质可及性，我们确定了 I 型和 II 型特异性效应的可能分子机制，区分细胞质信号网络的调节和协同调节巨噬细胞免疫反应的核表观基因组。
Macrophages orchestrate immune responses by sensing and responding to pathogen-associated molecules. These responses are modulated by prior conditioning with cytokines such as interferons (IFNs). Type I and II IFN have opposing functions in many biological scenarios, yet macrophages directly stimulated with Type I or II IFN activate highly overlapping gene expression programs. We hypothesized that a sequential conditioning-stimulation approach would reveal with greater specificity the differential effects of Type I and II IFN on human macrophages. By first conditioning with IFN then stimulating with toll-like receptor ligands and cytokines, followed by genome-wide RNA-seq analysis, we identified 713 genes whose expression was unaffected by IFN alone but showed potentiated or diminished responses to a stimulus after conditioning. For example, responses to the cytokine TNF were restricted by Type II IFN conditioning but potentiated by Type I IFN conditioning. We observed that the effects of IFN were not uniformly pro- or anti-inflammatory, but highly gene-specific and stimulus-specific. By assessing expression levels of key signal transducers and characterizing chromatin accessibility by ATAC-seq, we identify the likely molecular mechanisms underlying Type I and Type II-specific effects, distinguishing between modulation of cytoplasmic signaling networks and the nuclear epigenome that synergistically regulate macrophage immune responses.