在HBV转染的细胞中鉴定IFN-λ3调节的抗病毒蛋白的综合蛋白质

StephenW

Mol Cell Proteomics. 2018 Aug 10. pii: mcp.RA118.000735. doi: 10.1074/mcp.RA118.000735. [Epub ahead of print]
Comprehensive proteomics identification of IFN-λ3-regulated anti-viral proteins in HBV-transfected cells.
Makjaroen J1, Somparn P2, Hodge K3, Poomipak W4, Hirankarn N4, Pisitkun T5.
Author information

1
    Faculty of Medicine, Chulalongkorn University, Thailand.
2
    Chulalongkorn University.
3
    Mahidol University.
4
    Chulalongkorn University, Thailand.
5
    Center of Excellence in Systems Biology, Research affairs, Faculty of Medicine, Chulalongkorn University, Thailand [email protected].

Abstract

Interferon lambda (IFN-λ) is a relatively unexplored, yet promising anti-viral agent. IFN-λ has recently been tested in clinical trials of chronic hepatitis B virus infection (CHB), with the advantage that side effects may be limited compared with IFN-α, as IFN-λ receptors are found only in epithelial cells. To date, IFN-λ's downstream signaling pathway remains largely unelucidated, particularly via proteomics methods. Here, we report that IFN-λ3 inhibits HBV replication in HepG2.2.15 cells, reducing levels of both HBV transcripts and intracellular HBV DNA. Quantitative proteomic analysis of HBV-transfected cells was performed following 24-hour IFN-λ3 treatment, with parallel IFN-α2a and PBS treatments for comparison using a dimethyl labeling method. The depth of the study allowed us to map the induction of anti-viral proteins to multiple points of the viral life cycle, as well as facilitating the identification of anti-viral proteins not previously known to be elicited upon HBV infection (e.g. IFITM3, XRN2, and NT5C3A). This study also shows up-regulation of many effectors involved in antigen processing/presentation indicating that this cytokine exerted immunomodulatory effects through a number of essential molecules for these processes. Interestingly, the 2 subunits of the immunoproteasome cap (PSME1 and PSME2) were up-regulated while cap components of the constitutive proteasome were down-regulated upon both IFN treatments, suggesting coordinated modulation towards the antigen processing/presentation mode. Furthermore, in addition to confirming canonical activation of interferon-stimulated gene (ISG) transcription through the JAK-STAT pathway, we reveal that IFN-λ3 restored levels of RIG-I and RIG-G, proteins known to be suppressed by HBV. Enrichment analysis demonstrated that several biological processes including RNA metabolism, translation, and ER-targeting were differentially regulated upon treatment with IFN-λ3 vs. IFN-α2a. Our proteomic data suggests that IFN-λ3 regulates an array of cellular processes to control HBV replication.
KEYWORDS:

Immunology*; Mass Spectrometry; Quantification; RIG-I; Systems biology*; Viruses; dimethyl labeling; immunoproteasome; quantitative proteomics; type III IFN

PMID:
    30097535
DOI:
    10.1074/mcp.RA118.000735

StephenW

Mol细胞蛋白质组学。 2018年8月10日.pii：mcp.RA118.000735。 doi：10.1074 / mcp.RA118.000735。 [提前打印]
在HBV转染的细胞中鉴定IFN-λ3调节的抗病毒蛋白的综合蛋白质组学。
Makjaroen J1，Somparn P2，Hodge K3，Poomipak W4，Hirankarn N4，Pisitkun T5。
作者信息

1
    泰国朱拉隆功大学医学院。
2
    朱拉隆功大学。
3
    玛希隆大学。
4
    泰国朱拉隆功大学。
五
    泰国朱拉隆功大学医学院研究事务系统生物学卓越中心[email protected]。

抽象

干扰素λ（IFN-λ）是一种相对未开发但尚未发挥作用的抗病毒剂。最近在慢性乙型肝炎病毒感染（CHB）的临床试验中测试了IFN-λ，其优点是与IFN-α相比可能限制副作用，因为IFN-λ受体仅在上皮细胞中发现。迄今为止，IFN-λ的下游信号传导途径仍然很大程度上是不明确的，特别是通过蛋白质组学方法。在这里，我们报告IFN-λ3抑制HepG2.2.15细胞中的HBV复制，降低HBV转录物和细胞内HBV DNA的水平。在24小时IFN-λ3处理后进行HBV转染细胞的定量蛋白质组学分析，使用平行IFN-α2a和PBS处理，使用二甲基标记方法进行比较。研究的深度使我们能够将抗病毒蛋白的诱导图谱映射到病毒生命周期的多个点，并促进鉴定以前未知在HBV感染后引起的抗病毒蛋白（例如IFITM3，XRN2）和NT5C3A）。该研究还显示参与抗原加工/呈递的许多效应子的上调，表明该细胞因子通过许多必需分子对这些过程发挥免疫调节作用。有趣的是，免疫蛋白酶体上限（PSME1和PSME2）的2个亚基上调，而组成型蛋白酶体的上限组分在两种IFN处理中均下调，表明对抗原加工/呈递模式的协调调节。此外，除了通过JAK-STAT途径确认干扰素刺激基因（ISG）转录的规范激活外，我们还发现IFN-λ3恢复了已知被HBV抑制的蛋白质RIG-I和RIG-G的水平。富集分析表明，在用IFN-λ3对IFN-α2a处理后，包括RNA代谢，翻译和ER靶向的几种生物学过程被差异调节。我们的蛋白质组学数据表明，IFN-λ3调节一系列细胞过程以控制HBV复制。
关键词：

免疫学*;质谱;定量; RIG-I;系统生物学*;病毒;二甲基标记;免疫蛋白酶;定量蛋白质组学; III型IFN

结论：
    30097535
DOI：
    10.1074 / mcp.RA118.000735