牛痘病毒感染人巨噬细胞基因表达谱的RNA-Seq分析

RNA-Seq analysis of gene expression profile of human macrophages infected by vaccinia virus

摘要:

[目的] 利用RNA-Seq，分析人巨噬细胞在牛痘病毒（VACV）感染前后基因表达的变化，探索牛痘病毒与宿主细胞相互作用的机制。[方法] 用牛痘病毒感染人巨噬细胞，采用RNA-Seq比较感染组和对照组的差异表达基因，并进行KEGG、GO以及STRING网络分析。[结果] 感染组与对照组相比，筛选出显著性差异表达基因4796个，其中上调表达2416个，下调表达2380个。KEGG富集分析表明差异基因主要参与新陈代谢、信号转导、免疫系统和感染疾病等通路。GO功能注释显示这些基因富集在细胞功能调控、物质代谢和免疫调控等生命过程。运用STRING在线蛋白互作数据库，对NOD样信号通路进行分析，鉴定出以JUN、CHUK、IL1B和PYCARD 为核心的调控基因。[结论] 牛痘病毒感染可以诱导人巨噬细胞基因差异性表达，涉及的生物学过程有很多，对免疫相关的信号通路进行深入的分析，发现C型凝集素受体信号通路（C-type lectin receptor signaling pathway）、Toll样受体信号通路以及NOD样通路等多条通路可能参与调控牛痘病毒感染引起的炎症反应，该研究为解析牛痘病毒的感染机制和免疫逃逸机制以及其在临床上治疗感染性疾病和癌症提供了新思路。

Abstract:

[Objective] In this study, we used high throughput sequencing (RNA-Seq) to investigate the gene expression profiles in human monocyte-derived macrophages (THP-1) before and post-infection of vaccinia virus (VACV).[Methods] We analyzed differentially expressed genes in human macrophages with and without vaccinia virus infection by RNA-seq and performed the KEGG, GO, and STRING network analysis to study the immune-related signal alterations during infection.[Results] A total of 4796 genes were differentially expressed (2416 genes upregulated and 2380 genes downregulated) of VACV treatment THP-1 versus control. Further KEGG enrichment analysis shows that differential genes expression in VACV treatment THP-1 cells were mainly involved in metabolism, signal transduction, immune system, infectious diseases and other pathways. Gene Ontology (GO) functional annotation indicated that these genes were mainly enriched on cell function regulation, metabolism, immune regulation and other life processes. The STRING online database analysis showed that JUN, CHUK, IL1B, PYCARD protein were higher in the infected THP-1 cells. [Conclusion] Vaccinia virus can induce the differential expression of various genes in host cells, and multiple biological processes. We conducted in-depth analysis of immune-related signal pathways, and found the C-type lectin receptor-, NOD-like receptor-, and Toll-like receptor- signaling pathway are involved in inflammation responses induced by vaccinia virus infection. These results provide new insights not only in exploring the mechanism of interaction between vaccinia virus and host, but also in applying it in treatments of infectious diseases and cancers.