Transcriptome analyses of Atlantic salmon (Salmo salar L.) erythrocytes infected with piscine orthoreovirus (PRV)
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ABSTRACT:
ORGANISM(S): Salmo salar
PROVIDER: GSE183005 | GEO | 2021/08/30
REPOSITORIES: GEO
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Project description:Transcriptome analyses of Atlantic salmon (Salmo salar L.) erythrocytes infected with piscine orthoreovirus (PRV)
| PRJNA758707 | ENA
Project description:Piscine orthoreovirus (PRV) is a reovirus that has predominantly been detected in Atlantic salmon (Salmo salar L.). PRV is associated with heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon, and recently erythrocytes were identified as major target cells. The study of PRV replication and pathogenesis of the infection has been impeded by the inability to propagate PRV in vitro. In this study we developed an ex vivo cultivation system for PRV in Atlantic salmon erythrocytes. PRV was successfully passaged to naïve erythrocytes using lysates of blood cells from infected salmon. During cultivation a significant increase in viral load was observed by RT-qPCR and flow cytometry, which coincided with the formation of cytoplasmic inclusions. The inclusions resembled viral factories and contained both PRV protein and dsRNA. In addition, the erythrocytes generated an antiviral immune gene activation after PRV infection, with significant up-regulation of IFN-α, RIG-I, Mx and PKR transcripts. Supernatants from the first passage successfully transmitted virus to naïve erythrocytes. This study demonstrates that PRV replicates in Atlantic salmon erythrocytes ex vivo. The ex vivo infection model closely reflects the situation in vivo and can be used to study the infection and replication mechanisms of PRV, as well as the antiviral immune responses of salmonid erythrocytes.
| S-EPMC4350956 | biostudies-literature
Project description:PD and HSMI are viral diseases that cause heavy damages in Atlantic salmon aquaculture. This study was performed to examine and compare the time-courses of transcriptome responses to the causative agents - salmon alphavirus (SAV) and piscine reovirus (PRV).
2021-09-03 | GSE183260 | GEO
Project description:BackgroundCardiomyopathy syndrome (CMS) is a severe disease affecting large farmed Atlantic salmon. Mortality often appears without prior clinical signs, typically shortly prior to slaughter. We recently reported the finding and the complete genomic sequence of a novel piscine reovirus (PRV), which is associated with another cardiac disease in Atlantic salmon; heart and skeletal muscle inflammation (HSMI). In the present work we have studied whether PRV or other infectious agents may be involved in the etiology of CMS.ResultsUsing high throughput sequencing on heart samples from natural outbreaks of CMS and from fish experimentally challenged with material from fish diagnosed with CMS a high number of sequence reads identical to the PRV genome were identified. In addition, a sequence contig from a novel totivirus could also be constructed. Using RT-qPCR, levels of PRV in tissue samples were quantified and the totivirus was detected in all samples tested from CMS fish but not in controls. In situ hybridization supported this pattern indicating a possible association between CMS and the novel piscine totivirus.ConclusionsAlthough causality for CMS in Atlantic salmon could not be proven for either of the two viruses, our results are compatible with a hypothesis where, in the experimental challenge studied, PRV behaves as an opportunist whereas the totivirus might be more directly linked with the development of CMS.
| S-EPMC2994541 | biostudies-literature
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.
| S-EPMC4234517 | biostudies-literature
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
2007-12-03 | GSE8676 | GEO
Project description:Piscine orthoreovirus (PRV-1) can cause heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon (Salmo salar). The virus targets erythrocytes in the acute peak phase, followed by cardiomyocytes, before the infection subsides into persistence. The persistent phase is characterized by high level of viral RNA, but low level of viral protein. The origin and nature of persistent PRV-1 are not clear. Here, we analyzed for viral persistence and activity in various tissues and cell types in experimentally infected Atlantic salmon. Plasma contained PRV-1 genomic dsRNA throughout an 18-week long infection trial, indicating that viral particles are continuously produced and released. The highest level of PRV-1 RNA in the persistent phase was found in kidney. The level of PRV-1 ssRNA transcripts in kidney was significantly higher than that of blood cells in the persistent phase. In-situ hybridization assays confirmed that PRV-1 RNA was present in erythroid progenitor cells, erythrocytes, macrophages, melano-macrophages and in some additional un-characterized cells in kidney. These results show that PRV-1 establishes a productive, persistent infection in Atlantic salmon and that erythrocyte progenitor cells are PRV target cells.
| S-EPMC6784031 | biostudies-literature
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
2007-12-01 | GSE9259 | GEO
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.
2010-06-25 | E-GEOD-9259 | biostudies-arrayexpress
Project description:Piscine orthoreovirus (PRV) is associated with heart- and skeletal muscle inflammation (HSMI) of farmed Atlantic salmon (Salmo salar). We have performed detailed sequence analysis of the PRV genome with focus on putative encoded proteins, compared with prototype strains from mammalian (MRV T3D)- and avian orthoreoviruses (ARV-138), and aquareovirus (GCRV-873). Amino acid identities were low for most gene segments but detailed sequence analysis showed that many protein motifs or key amino acid residues known to be central to protein function are conserved for most PRV proteins. For M-class proteins this included a proline residue in ?2 which, for MRV, has been shown to play a key role in both the formation and structural organization of virus inclusion bodies, and affect interferon-? signaling and induction of myocarditis. Predicted structural similarities in the inner core-forming proteins ?1 and ?2 suggest a conserved core structure. In contrast, low amino acid identities in the predicted PRV surface proteins ?1, ?1 and ?3 suggested differences regarding cellular interactions between the reovirus genera. However, for ?1, amino acid residues central for MRV binding to sialic acids, and cleavage- and myristoylation sites in ?1 required for endosomal membrane penetration during infection are partially or wholly conserved in the homologous PRV proteins. In PRV ?3 the only conserved element found was a zinc finger motif. We provide evidence that the S1 segment encoding ?3 also encodes a 124 aa (p13) protein, which appears to be localized to intracellular Golgi-like structures. The S2 and L2 gene segments are also potentially polycistronic, predicted to encode a 71 aa- (p8) and a 98 aa (p11) protein, respectively. It is concluded that PRV has more properties in common with orthoreoviruses than with aquareoviruses.
| S-EPMC3726481 | biostudies-literature