Transcriptome analysis of avian reovirus-mediated changes in gene expression of normal chicken fibroblast DF-1 cells

Abstract

Background: Avian reovirus (ARV) is an important poultry pathogen that can cause immunosuppression. In this study, RNA-Seq technology was applied to investigate the transcriptome-wide changes of DF-1 cells upon ARV infection at the middle stage. Results: Total RNA of ARV-infected or mock-infected samples at 10 and 18 h post infection (hpi) was extracted to build RNA-Seq datasets. Analysis of the sequencing data revealed that the expressions of numerous genes were altered, and a panel of differentially expressed genes were confirmed with RT-qPCR. At 10 hpi, 104 genes were down-regulated and 64 were up-regulated, while the expressions of 47 genes were increased and only one was down-regulated, which may play a role in retinoic acid biosynthesis, at 18 hpi in the ARV-infected cells. The similar profiles of up-regulated genes between the two groups of infected cells suggest that ARV infection activated a prolonged antiviral response of host cells. Alternative splicing analysis found no significantly changed events altered by ARV infection. Conclusions: Overall, the differential expression profile presented in this study can be used to expand our understanding of the comprehensive interactions between ARV and the host cells, and may be helpful for us to reveal the pathogenic mechanism on the molecular level.

In vitro study on role of σB protein in avian reovirus pathogenesis

“…Most recently a report identified that Wnt14 plays a pivotal role in initiating synovial joint formation in the chick limb, but the researchers were unable to determine the specific pathway that is responsible for the joint pathogenesis of ARV [ 39 ]. It was shown that the elevated expression of Wnt14 gene was observed, which might play a key role in the development of the disease [ 33 ]. The detailed study regarding the molecular arthritic pathogenesis and role of σB protein in viral arthritis was still unknown.…”

“…Identification and complete genome sequencing of two naturally occurring ARV variant strain co-infections having same M2 segment but were distantly evolved nine other segments in layer chickens using NGS was achieved [ 32 ]. Most recently, a gene expression RNA-Seq study showed that inoculation of chicken fibroblast DF-1 cells lines with ARV stimulates a prolonged antiviral response in host cells and interferes with cell growth and cell death pathways [ 33 ]. Despite many studies towards its diagnosis and pathogenesis induced by ARV, no study signifies the role of various genes of arthritis pathway expression in ARV and σB protein induced pathogenesis, which was the main objective of our study.…”

Transcriptome analysis of postharvest blueberries (Vaccinium corymbosum ‘Duke’) in response to cold stress

“…Total RNAs from all samples were isolated using the Omi-niPlant RNA kit (CWBio, Beijing, China). The integrity was monitored on 1% agarose gels and the purity was checked using the NanoDrop spectrophotometer (Thermo Fisher Scientific, Inc.) according to Niu et al (2017) [76]. Sequencing libraries were generated using NEBNext®Ultra™ RNA Library Prep Kit (Illumina®NEB, USA) and index codes were added to attribute sequences to each sample.…”

Cholesterol-Rich Lipid Rafts in the Cellular Membrane Play an Essential Role in Avian Reovirus Replication

“…Vero cells were maintained in Dulbecco's modified Eagle's medium (DMEM; Gibco, Shanghai, China) supplemented with 10% fetal bovine serum (FBS) (Gibco) at 37 • C with 5% CO 2 . The ARV strain GX/2010/1 was isolated by our laboratory and was propagated in chicken embryo fibroblast (CEF) cells (Meng et al, 2012;Duan et al, 2015;Niu et al, 2017Niu et al, , 2019.…”