Project description:Piscine reovirus (PRV) is a causative agent of heart and skeletal muscle inflammation in Atlantic salmon, which is propagated in red blood cells (RBC). Here, transcriptome analyses of PRV infected erythrocytes showed strong and complex innate antiviral responses.
Project description:Circular RNAs (circRNAs) and microRNAs (miRNAs) participate in regulating many biological processes. However, their roles in PrV-II pathogenicity are largely unknown. Here, we analyzed the expression profile of circRNAs and miRNAs in the PrV-DX, a wild-type (WT) strain of PRV-II, and its attenuated gE-TK- PRV-DX infected cells by high-throughput sequencing.
Project description:To study in vitro the epithelial cells and PrV interactions during infection, we followed PrV and PK15 cells transcriptome modifications during time-course infection (I) and mock-infection (MI).Six time points were studied: just after I and MI, 1h, 2h, 4h, 8h and 12h post-I and MI. For this study, a pig DNA/cDNA microarray containing genes of the SLA region, additional genes encoding other important immunological molecules and all the PrV genes was constructed. Keywords: infection time course
Project description:To study in vitro the epithelial cells and PrV interactions during infection, we followed PrV and PK15 cells transcriptome modifications during time-course infection (I) and mock-infection (MI). Four time points were studied: 1h, 2h, 4h and 8h post-I and MI. Four replicates of I and MI were analysed. 32 samples - The hybridization scheme which can be define as dye-switch was chosen. A balanced loop design with two independent loops, each loop containing 2 replicates of PrV infection and MI, was used. In total, 32 slides were used in this experiment.
Project description:To study in vitro the epithelial cells and PrV interactions during infection, we followed PrV and PK15 cells transcriptome modifications during time-course infection (I) and mock-infection (MI). Four time points were studied: 1h, 2h, 4h and 8h post-I and MI. Four replicates of I and MI were analysed. Keywords: Pig, PrV, Pk15 cells, kinetics
Project description:Purpose: Evaluation of the m6A modification of PRV and PK15 transcripts during PRV infection Methods: Porcine kidney cell line PK15 was uninfected or infected with PRV for 24 hours. Total RNA from each sample were extracted. Intact mRNA was isolated from total RNA samples and then chemically fragmented to 300-nucleoside-long fragments. Fragmented mRNAs were immunoprecipitated with anti-N6-methyadenosine (m6A) antibody (a part of the fragmented mRNAs was kept as input). Both m6A enriched mRNAs and input mRNAs were concentrated for RNA-seq libraries construction. The libraries were forwarded to sequencing run on Illumina NovaSeq 6000. Results: PRV transcripts were m6A modified during PRV infection and PRV infection changed m6A modification profiles of PK15 transcripts.
Project description:Little is known about the latent infection period and lytic infection mechanism of pseudorabies virus. The advent of high-throughput sequencing technologies has allowed us to examine many different biological components simultaneously. To examine the effects of glucocorticoids on latently infected PRV neurons, we constructed gene expression profiles of RNA-seq data for miRNA expression from PC-12 cell with a PRV latent infection model group (PRV group), a reactivation model group (DEX group) and a blank control group (CON group).
Project description:Purpose: Next Generation sequencing (NGS) has revolutionized system-based cellular pathway analysis. The aim of this study was to infect differentially expressed genes for the BV2 phenotype with cur-transformed PRV and to predict possible molecular mechanisms involved in phenotypic transformation.Methods: The BV2 cells in the three treatment groups were sequenced by Illumina sequencing platform, and three repeated mRNA maps of BV2 cells were obtained. The cells were divided into control group: normal BV2 cells were cultured for 48 hours; PRV infection group: BV2 cells were infected with 1.66×106TCID50 PRV for 48h; CUR treatment group: BV2 cells were infected with 1.66×106TCID50 PRV for 24h and then treated with 20μM CUR for 24h.Results:The number of differentially expressed genes in PRV group and CON group was 306, including 242 biologically significant up-regulated genes and 64 biologically significant down-regulated genes. The number of differentially expressed genes in PRV+CUR group and PRV group was 5073, among which 2661 were biologically significant up-regulated genes and 2412 were biologically significant down-regulated genes. Compared with CON group, 1785 differentially expressed genes were found in PRV+CUR group, including 788 biologically significant up-regulated genes and 997 biologically significant down-regulated genes.Conclusions: Bioinformatics analysis of transcriptome characteristics and differentially expressed genes of BV2 in different phenotypes after PRV infection and CUR treatment showed that glycolysis, oxidative phosphorylation, Alzheimer's disease and fatty acid synthesis pathway may be involved in the process of BV2 activation into M1 type. Glycolysis, gluconeogenesis, insulin signaling pathway, NF-κB pathway, AMPK pathway, oxidative phosphorylation and fatty acid synthesis may be involved in the phenotypic transformation of PRV-infected BV2 with CUR. Among them, AMPK may influence phenotypic transformation by controlling the transformation of energy metabolism.
Project description:Piscine orthoreovirus (PRV) belongs to the Reoviridae family and is the only known fish virus related to the Orthoreovirus genus. The virus is the causative agent of heart and skeletal muscle inflammation (HSMI), an emerging disease in farmed Atlantic salmon (Salmo salar L.). PRV is ubiquitous in farmed Atlantic salmon and high loads of PRV in the heart are consistent findings in HSMI. The mechanism by which PRV infection causes disease remains largely unknown. In this study we investigated the presence of PRV in blood and erythrocytes using an experimental cohabitation challenge model. We found that in the early phases of infection, the PRV loads in blood were significantly higher than in any other organ. Most virus was found in the erythrocyte fraction, and in individual fish more than 50% of erythrocytes were PRV-positive, as determined by flow cytometry. PRV was condensed into large cytoplasmic inclusions resembling viral factories, as demonstrated by immunofluorescence and confocal microscopy. By electron microscopy we showed that these inclusions contained reovirus-like particles. The PRV particles and inclusions also had a striking resemblance to previously reported viral inclusions described as Erythrocytic inclusion body syndrome (EIBS). We conclude that the erythrocyte is a major target cell for PRV infection. These findings provide new information about HSMI pathogenesis, and show that PRV is an important factor of viral erythrocytic inclusions.
Project description:Little is known about the latent infection period and lytic infection mechanism of pseudorabies virus. The advent of high-throughput sequencing technologies has allowed us to examine many different biological components simultaneously. To examine the effects of glucocorticoids on latently infected PRV neurons, we constructed gene expression profiles of RNA-seq data from PC-12 cell with a PRV latent infection model group (PRV group), a reactivation model group (DEX group) and a blank control group (CON group).