Fundamental to mammalian intrinsic and innate immune defenses against pathogens is the production of Type I and Type II interferons, such as IFN-β and IFN-γ, respectively. The comparative effects of IFN classes on the cellular proteome, protein interactions, and virus restriction within cell types that differentially contribute to immune defenses are needed for understanding immune signaling. Here, a multilayered proteomic analysis, paired with biochemical and molecular virology assays, allows distinguishing host responses to IFN-β and IFN-γ and associated antiviral impacts during infection with several ubiquitous human viruses. In differentiated macrophage-like monocytic cells, we classified proteins upregulated by IFN-β, IFN-γ, or pro-inflammatory LPS. Using parallel reaction monitoring, we developed a proteotypic peptide library for shared and unique ISG signatures of each IFN class, enabling orthogonal confirmation of protein alterations. Thermal proximity coaggregation analysis identified the assembly and maintenance of IFN-induced protein interactions. Comparative proteomics and cytokine responses in macrophage-like monocytic cells and primary keratinocytes provided contextualization of their relative capacities to restrict virus production during infection with herpes simplex virus type-1, adenovirus, and human cytomegalovirus. Our findings demonstrate how IFN classes induce distinct ISG abundance and interaction profiles that drive antiviral defenses within cell types that differentially coordinate mammalian immune responses.

中文翻译：

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干扰素类别对宿主炎症反应和限制病毒后代产生的不同贡献

哺乳动物针对病原体的内在和先天免疫防御的基础是 I 型和 II 型干扰素的产生，分别如 IFN-β 和 IFN-γ。为了理解免疫信号传导，需要比较干扰素类别对细胞蛋白质组、蛋白质相互作用和细胞类型内病毒限制的影响，这些因素对免疫防御有不同的贡献。在这里，多层蛋白质组分析与生化和分子病毒学检测相结合，可以区分宿主对 IFN-β 和 IFN-γ 的反应以及在感染几种普遍存在的人类病毒期间相关的抗病毒影响。在分化的巨噬细胞样单核细胞中，我们对 IFN-β、IFN-γ 或促炎性 LPS 上调的蛋白质进行了分类。通过平行反应监测，我们开发了一个蛋白质肽库，用于每个 IFN 类别的共享和独特的 ISG 特征，从而能够正交确认蛋白质改变。热邻近共聚集分析确定了 IFN 诱导的蛋白质相互作用的组装和维持。巨噬细胞样单核细胞和原代角质形成细胞中的比较蛋白质组学和细胞因子反应提供了它们在 1 型单纯疱疹病毒、腺病毒和人巨细胞病毒感染期间限制病毒产生的相对能力的背景。我们的研究结果表明，IFN 类别如何诱导不同的 ISG 丰度和相互作用特征，从而驱动不同协调哺乳动物免疫反应的细胞类型内的抗病毒防御。