Transcriptional profiling of feline infectious peritonitis virus infection in CRFK cells and in PBMCs from FIP diagnosed cats
Transcriptional profiling of feline infectious peritonitis virus infection in CRFK cells and in PBMCs from FIP diagnosed cats
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DOI：
10.1186/1743-422x-10-329
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发表时间：
2013-11-09
期刊：
影响因子：
4.8
通讯作者：
Omar, Abdul Rahman
中科院分区：
文献类型：
--
作者：
Harun, Mohammad Syamsul Reza;Kuan, Choong Oi;Omar, Abdul Rahman
背景：猫传染性腹膜炎（FIP）是由FIP病毒（FIPV）引起的一种致死性全身性疾病；猫肠道冠状病毒（FECV）的一种毒力突变体。目前，FIPV感染过程中病毒毒力决定因素和宿主基因表达尚不完全清楚。方法：采用Illumina下一代测序方法，对感染FIPV菌株79-1146后3小时感染的CRFK细胞进行RNA测序。Bioinformatic的分析基于Felis catus 2X注释鸟枪参考基因组，使用CLC bio genome Workbench将对照和感染细胞的读取图谱绘制为19046个注释基因中的18899个基因。采用Kal's Z检验统计分析感染CRFK细胞的差异表达基因。采用实时RT-qPCR技术对健康猫和fip病猫感染的CRFK细胞和外周血单个核细胞（PBMCs）中的PD-1、PD-L1和A3H三个基因进行了进一步的转录分析。结果：根据Kal's z检验，在基因表达的错误发现率（FDR）变化为1.99倍的情况下，两种样本共有61个基因差异表达，其中44个基因上调，其余基因下调。大多数基因紧密地聚集在一起，表明表达是同质的。大部分被显著调控的基因是与单核-巨噬细胞和Th1细胞功能相关的基因，以及与凋亡调控相关的基因。在FIPV感染的CRFK细胞以及健康猫和FIP诊断猫的pbmc中，实时RT-qPCR研究了2个上调基因（PD-L1和A3H）以及凋亡相关基因PD-1的表达，结果与转录组数据一致。结论：讨论了这些基因在猫冠状病毒感染中的可能作用及其重要性。
Background: Feline Infectious Peritonitis (FIP) is a lethal systemic disease, caused by the FIP Virus (FIPV); a virulent mutant of Feline Enteric Coronavirus (FECV). Currently, the viruses virulence determinants and host gene expressions during FIPV infection are not fully understood.Methods: RNA sequencing of Crandell Rees Feline Kidney (CRFK) cells, infected with FIPV strain 79-1146 at 3 hours post infection (h.p.i), were sequenced using the Illumina next generation sequencing approach. Bioinformatic's analysis, based on Felis catus 2X annotated shotgun reference genome, using CLC bio Genome Workbench mapped both control and infected cell reads to 18899 genes out of 19046 annotated genes. Kal's Z test statistical analysis was used to analyse the differentially expressed genes from the infected CRFK cells. Real time RT-qPCR was developed for further transcriptional profiling of three genes (PD-1, PD-L1 and A3H) in infected CRFK cells and Peripheral Blood Mononuclear Cells (PBMCs) from healthy and FIP-diseased cats.Results: Based on Kal's Z-test, with False Discovery Rate (FDR) 1.99 fold change on gene expressions, a total of 61 genes were differentially expressed by both samples, where 44 genes were up-regulated and the remainder were down-regulated. Most genes were closely clustered together, suggesting a homogeneous expression. The majority of the genes that were significantly regulated, were those associated with monocytes-macrophage and Th1 cell functions, and the regulation of apoptosis. Real time RT-qPCR developed focusing on 2 up-regulated genes (PD-L1 and A3H) together with an apoptosis associated gene PD-1 expressions in FIPV infected CRFK cells and in PBMCs from healthy and FIP diagnosed cats produced concordant results with transcriptome data.Conclusion: The possible roles of these genes, and their importance in feline coronaviruses infection, are discussed.