The major immediate early 62 (IE62) protein of varicella-zoster virus (VZV) is delivered to newly infected cell nuclei, where it initiates VZV replication by transactivating viral immediate early (IE), early (E), and late (L) genes. Interferon gamma (IFN-γ) is a potent cytokine produced following primary VZV infection. Furthermore, VZV reactivation correlates with a decline in IFN-γ-producing immune cells. Our results showed that treatment with 20 ng/ml of IFN-γ completely reduced intracellular VZV yield in A549 lung epithelial cells, MRC-5 lung fibroblasts, and ARPE-19 retinal epithelial cells at 4 days post-VZV infection. However, IFN-γ reduced virus yield only 2-fold in MeWo melanoma cells compared to that of untreated cells. IFN-β significantly inhibited VZV replication in both ARPE-19 and MeWo cells. In luciferase assays with VZV open reading frame 61 (ORF61) promoter reporter plasmid, IFN-γ abrogated the transactivation activity of IE62 by 95%, 97%, and 89% in A549, ARPE-19, and MRC-5 cells, respectively. However, IFN-γ abrogated IE62's transactivation activity by 16% in MeWo cells, indicating that IFN-γ inhibits VZV replication as well as IE62-mediated transactivation in a cell line-dependent manner. The expression of VZV IE62 and ORF63 suppressed by IFN-γ was restored by JAK1 inhibitor treatment, indicating that the inhibition of VZV replication is mediated by JAK/STAT1 signaling. In the presence of IFN-γ, knockdown of interferon response factor 1 (IRF1) increased VZV replication. Ectopic expression of IRF1 reduced VZV yields 4,000-fold in MRC-5 and ARPE-19 cells but 3-fold in MeWo cells. These results suggest that IFN-γ blocks VZV replication by inhibiting IE62 function in a cell line-dependent manner.IMPORTANCE Our results showed that IFN-γ significantly inhibited VZV replication in a cell line-dependent manner. IFN-γ inhibited VZV gene expression after the immediate early stage of infection and abrogated IE62-mediated transactivation. These results suggest that IFN-γ blocks VZV replication by inhibiting IE62 function in a cell line-dependent manner. Understanding the mechanisms by which IFN-γ plays a role in VZV gene programming may be important in determining the tissue restriction of VZV.

中文翻译：

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干扰素γ以细胞系依赖性方式抑制水痘-带状疱疹病毒的复制。

水痘带状疱疹病毒（VZV）的主要立即早期62（IE62）蛋白被递送至新感染的细胞核，在那里它通过激活病毒早期（IE），早期（E）和晚期（L）基因来启动VZV复制。 。干扰素γ（IFN-γ）是原发性VZV感染后产生的强力细胞因子。此外，VZV再激活与产生IFN-γ的免疫细胞下降有关。我们的结果表明，在VZV感染后4天，用20 ng / mlIFN​​-γ的治疗完全降低了A549肺上皮细胞，MRC-5肺成纤维细胞和ARPE-19视网膜上皮细胞的细胞内VZV产量。但是，与未经处理的细胞相比，在MeWo黑色素瘤细胞中，IFN-γ降低的病毒产量仅为2倍。IFN-β在ARPE-19和MeWo细胞中均显着抑制VZV复制。在使用VZV开放阅读框61（ORF61）启动子报告质粒的荧光素酶测定中，IFN-γ分别使A549，ARPE-19和MRC-5细胞中IE62的反式激活活性降低了95％，97％和89％。然而，IFN-γ在MeWo细胞中废除了IE62的反式激活活性16％，表明IFN-γ以细胞系依赖性方式抑制VZV复制以及IE62介导的反式激活。通过JAK1抑制剂处理恢复了被IFN-γ抑制的VZV IE62和ORF63的表达，表明对VZV复制的抑制作用是由JAK / STAT1信号传导介导的。在存在IFN-γ的情况下，干扰素反应因子1（IRF1）的敲低会增加VZV复制。IRF1的异位表达在MRC-5和ARPE-19细胞中降低了VZV产量4,000倍，在MeWo细胞中降低了3倍。这些结果表明，IFN-γ通过以细胞系依赖性方式抑制IE62功能来阻断VZV复制。重要结果我们的结果表明，IFN-γ以细胞系依赖性方式显着抑制VZV复制。在感染的早期后，IFN-γ抑制了VZV基因的表达，并废除了IE62介导的反式激活。这些结果表明，IFN-γ通过以细胞系依赖性方式抑制IE62功能来阻断VZV复制。理解IFN-γ在VZV基因编程中发挥作用的机制对于确定VZV的组织限制可能很重要。在感染的早期后，IFN-γ抑制了VZV基因的表达，并废除了IE62介导的反式激活。这些结果表明，IFN-γ通过以细胞系依赖性方式抑制IE62功能来阻断VZV复制。理解IFN-γ在VZV基因编程中发挥作用的机制对于确定VZV的组织限制可能很重要。在感染的早期后，IFN-γ抑制了VZV基因的表达，并废除了IE62介导的反式激活。这些结果表明，IFN-γ通过以细胞系依赖性方式抑制IE62功能来阻断VZV复制。理解IFN-γ在VZV基因编程中发挥作用的机制对于确定VZV的组织限制可能很重要。