Morphogenesis of Respiratory Syncytial Virus in a Green Monkey Kidney Cell Line (Vero)

Norrby, Erling; Marusyk, Halyna; Örvell, Claes

doi:10.1128/jvi.6.2.237-242.1970

Cited by 92 publications

(50 citation statements)

References 12 publications

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“…This indicates that the ISA virus does not belong to the family Arenaviridae, as suggested . The inclusions in the ISA virus may be explained as transverse sections through nucleocapsids giving an impression of several granules (Norrby, Marusyk & Orvell 1970;Palmer & Martin 1988). The high number of granules in each virus and the lack of longitudinal sections of the nucleocapsids contradict slightly such an explanation, unless the virus genome consists of several short segments.…”

Section: Discussionmentioning

confidence: 99%

Presence of infectious salmon anaemia virus (ISAV) in tissues of Atlantic salmon, Salmo salar L., collected during three separate outbreaks of the disease

Nylund

Hovland

Watanabe

et al. 1995

Journal of Fish Diseases

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. Infectious salmon anaemia (ISA) is a viral disease of farmed Atlantic salmon, Salmo salar L., in Norway. The enveloped virus particles (100nm) believed to be the causative agent of the disease have been observed budding from endothelial cells in heart blood vessels. However, it is not known if the virus propagates in endothelial cells in all tissues/organs, if other target cells exist or if material collected from different salmon farms with natural outbreaks of ISA contain the same virus particles. Salmon smolts from three hatcheries with no history of disease were taken into the laboratory and experimentally challenged with ISA collected from Atlantic salmon during natural outbreaks of the disease in three different fish farms outside Bergen. Norway. Tissues for TEM studies were Collected from: (1) organs that showed clinical signs of ISA (i.e. used in the diagnosis of the disease); (2) tissues believed to be important in transmission of the virus (integument, kidney, urinary bladder, gut and somatic muscle); and (3) hormone‐producing tissues (pituitary gland, saccus vasculosus, thymus, thyroid, ultimobranehial gland, gonad, head kidney, heart and ventral aorta). The same virus as that believed to be the causative agent of ISA was found in all tissues examined from the challenged fish, i.e. a multiorgan infection with the same virus present in salmon from all three fish farms. The virus particles are about 100 nm in diameter, consisting of a slightly pleomorphic unit membrane envelope within which are a number of granules about 10–12nm in diameter. The granules seemed to be arranged in two concentric circles (spheres). The virus was seen budding from the surface of endothelial cells in blood vessels/sinus only. However, the virus was found intracellularly in both endothelial cells and in leucocytes.

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Section: Discussionmentioning

confidence: 99%

Presence of infectious salmon anaemia virus (ISAV) in tissues of Atlantic salmon, Salmo salar L., collected during three separate outbreaks of the disease

Nylund

Hovland

Watanabe

et al. 1995

Journal of Fish Diseases

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“…The presence of cytoplasmic inclusion bodies in RSV-infected cells was first observed by electron microscopy (31). Inclusion bodies, made of aggregated nucleocapsids, have also been observed in the cytoplasm of cells infected with other paramyxoviruses.…”

Section: Structure Of Inclusion Bodies In Live and Fixed Cellsmentioning

confidence: 96%

Live-Cell Characterization and Analysis of a Clinical Isolate of Bovine Respiratory Syncytial Virus, Using Molecular Beacons

Santangelo

Nitin

LaConte

et al. 2006

J Virol

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Understanding viral pathogenesis is critical for prevention of outbreaks, development of antiviral drugs, and biodefense. Here, we utilize molecular beacons to directly detect the viral genome and characterize a clinical isolate of bovine respiratory syncytial virus (bRSV) in living cells. Molecular beacons are dual-labeled, hairpin oligonucleotide probes with a reporter fluorophore at one end and a quencher at the other; they are designed to fluoresce only when hybridizing to a complementary target. By imaging the fluorescence signal of molecular beacons, the spread of bRSV was monitored for 7 days with a signal-to-noise ratio of 50 to 200, and the measured time course of infection was quantified with a mathematical model for viral growth. We found that molecular beacon signal could be detected in single living cells infected with a viral titer of 2 ؋ 10 3.6 50% tissue culture infective doses/ml diluted 1,000 fold, demonstrating high detection sensitivity. Low background in uninfected cells and simultaneous staining of fixed cells with molecular beacons and antibodies showed high detection specificity. Furthermore, using confocal microscopy to image the viral genome in live, infected cells, we observed a connected, highly three-dimensional, amorphous inclusion body structure not seen in fixed cells. Taken together, the use of molecular beacons for active virus imaging provides a powerful tool for rapid viral infection detection, the characterization of RNA viruses, and the design of new antiviral drugs.

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“…A hallmark feature of RSV infection in epithelial cells is the formation of discrete collections of viral replication proteins that have been termed viral inclusion bodies (28). These cytoplasmic structures increase in size during the course of infection and have been shown to contain the N, P, M2-1, L, and M proteins (3,9,21).…”

mentioning

confidence: 99%

Respiratory Syncytial Virus Induces Host RNA Stress Granules To Facilitate Viral Replication

Lindquist¹,

Lifland

Utley

et al. 2010

J Virol

149

175

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Mammalian cell cytoplasmic RNA stress granules are induced during various conditions of stress and are strongly associated with regulation of host mRNA translation. Several viruses induce stress granules during the course of infection, but the exact function of these structures during virus replication is not well understood. In this study, we showed that respiratory syncytial virus (RSV) induced host stress granules in epithelial cells during the course of infection. We also showed that stress granules are distinct from cytoplasmic viral inclusion bodies and that the RNA binding protein HuR, normally found in stress granules, also localized to viral inclusion bodies during infection. Interestingly, we demonstrated that infected cells containing stress granules also contained more RSV protein than infected cells that did not form inclusion bodies. To address the role of stress granule formation in RSV infection, we generated a stable epithelial cell line with reduced expression of the Ras-GAP SH3 domain-binding protein (G3BP) that displayed an inhibited stress granule response. Surprisingly, RSV replication was impaired in these cells compared to its replication in cells with intact G3BP expression. In contrast, knockdown of HuR by RNA interference did not affect stress granule formation or RSV replication. Finally, using RNA probes specific for RSV genomic RNA, we found that viral RNA predominantly localized to viral inclusion bodies but a small percentage also interacted with stress granules during infection. These results suggest that RSV induces a host stress granule response and preferentially replicates in host cells that have committed to a stress response.Respiratory syncytial virus (RSV) is a leading cause of serious viral lower respiratory tract illness in infants and the elderly worldwide. The virus is a member of the Paramyxoviridae family, and the genome consists of a single-stranded, negativesense RNA molecule that encodes 11 proteins. The ribonucleoprotein complex necessary for transcription and replication includes the nucleoprotein (N), the phosphoprotein (P), and the large polymerase protein (L). M2-1 and M2-2 are accessory proteins that are involved in transcription and replication, respectively (7). The fusion (F) protein, attachment protein (G), and small hydrophobic (SH) protein are found on the surface of infectious virions, while the matrix (M) protein locates inside the virion particle. Two nonstructural proteins (NS1 and NS2) are expressed in the cytoplasm of infected cells and appear to act as interferon antagonists during infection (36). The mechanisms by which the virus replicates and assembles in infected epithelial cells are incompletely understood.A hallmark feature of RSV infection in epithelial cells is the formation of discrete collections of viral replication proteins that have been termed viral inclusion bodies (28). These cytoplasmic structures increase in size during the course of infection and have been shown to contain the N, P, M2-1, L, and M proteins (3,9,21). It is tho...

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Morphogenesis of Respiratory Syncytial Virus in a Green Monkey Kidney Cell Line (Vero)

Cited by 92 publications

References 12 publications

Presence of infectious salmon anaemia virus (ISAV) in tissues of Atlantic salmon, Salmo salar L., collected during three separate outbreaks of the disease

Presence of infectious salmon anaemia virus (ISAV) in tissues of Atlantic salmon, Salmo salar L., collected during three separate outbreaks of the disease

Live-Cell Characterization and Analysis of a Clinical Isolate of Bovine Respiratory Syncytial Virus, Using Molecular Beacons

Respiratory Syncytial Virus Induces Host RNA Stress Granules To Facilitate Viral Replication

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