Deep sequencing of the transcriptome from murine lung infected with H5N8 subtype avian influenza virus with combined substitutions I283M and K526R in PB2 gene
Deep sequencing of the transcriptome from murine lung infected with H5N8 subtype avian influenza virus with combined substitutions I283M and K526R in PB2 gene
复制标题
对感染 H5N8 亚型禽流感病毒的 PB2 基因中 I283M 和 K526R 组合取代的小鼠肺部转录组进行深度测序。
DOI：
10.1016/j.meegid.2020.104672
复制
发表时间：
2021-01-01
影响因子：
3.2
通讯作者：
Liu，Xiufan
中科院分区：
文献类型：
--
作者：
Chen，Sujuan;Wang，Xiao;Liu，Xiufan
H5 N8亚型高致病性禽流感病毒（HPAIV）对养禽业和公众健康构成巨大威胁。我们前期的研究表明，PB 2基因中283 M和526 R的协同作用是病毒在哺乳动物中高致病性的关键因素。然而，突变病毒的潜在致病机制仍不清楚。在此，使用RNA-seq方法分析了亲本r-JY病毒和JY-PB 2-I283 M-K526 R突变病毒感染小鼠肺后的总体宿主应答。我们发现，无论是数量和宿主差异表达基因（DEG）的表达水平在突变病毒感染的小鼠相比，亲本病毒组。GO和KEGG分析表明，DEG主要与天然免疫反应有关。特别是，PB 2-I283 M-K526 R突变强烈诱导细胞因子风暴相关基因的急剧表达，包括MX 1，CXCL 10和IFN-γ，通过qRT-PCR进行。通过肺组织病理学相关DEG的热图分析和体外细胞凋亡检测，我们还发现PB 2-I283 M-K526 R突变促进了细胞凋亡水平。综上所述，我们的数据表明H5 N8亚型HPAIV的PB 2-I283 M-K526 R加剧了先天免疫反应和细胞凋亡水平，这可能是突变体在哺乳动物中致病性增强的关键致病机制。
H5N8 subtype highly pathogenic avian influenza viruses (HPAIVs) pose a huge threat to poultry industry and general public health. Our previous study demonstrated that synergistic effect of 283M and 526R in PB2 gene was a critical factor for viral high pathogenicity in mammals. However, the potential pathogenic mechanism of the mutant virus is still unclear. Here, RNA-seq method was used to analyze the global host response of murine lungs after infecting with parental r-JY virus and JY-PB2-I283M-K526R mutant virus. We found that both amounts and the expression levels of host differentially expressed genes (DEGs) were higher in mutant virus-infected mice compared with the group of parental virus. Furthermore, the DEGs mainly related with innate immune response by GO and KEGG analysis. Especially, PB2-I283M-K526R mutation strongly induced a sharp expression of cytokine storm-related genes, including MX1, CXCL10, and IFN-γ, performed by qRT-PCR. We also found that PB2-I283M-K526R mutation accelerated the level of cell apoptosis by heat map analysis of apoptosis-related DEGs in lungs and apoptosis assayin vitro. Taken together, our data demonstrated that PB2-I283M-K526R of H5N8 subtype HPAIV exacerbated the innate immune response and the level of cell apoptosis, which might be a key pathogenic mechanism for the enhanced pathogenicity of mutants in mammals.