Propagation of Swine Hemagglutinating Encephalomyelitis Virus and Pseudorabies Virus in Dorsal Root Ganglia Cells

Hara, Yoko; Hasebe, Rie; Sunden, Yuji; Ochiai, Kenji; Honda, Eiichi; Sakoda, Yoshihiro; Umemura, Takashi

doi:10.1292/jvms.71.595

Cited by 22 publications

(20 citation statements)

References 28 publications

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“…It is not known, however, if rearrangement of actin is required for early stages on alphaherpesvirus infection in vivo . In Vero cells and cultured sensory neurons, actin-depolymerization drugs have no impact on virus entry [143,184]. Conversely, the microtubule cytoskeleton and long-range microtubule-dependent transport are very important for alphaherpesvirus infection, especially in regard to axonal transport.…”

Section: Alphaherpesviruses and The Host Cell Cytoskeleton: Interactimentioning

confidence: 99%

Alphaherpesviruses and the Cytoskeleton in Neuronal Infections

Zaichick

Bohannon

Smith

2011

Viruses

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Following infection of exposed peripheral tissues, neurotropic alphaherpesviruses invade nerve endings and deposit their DNA genomes into the nuclei of neurons resident in ganglia of the peripheral nervous system. The end result of these events is the establishment of a life-long latent infection. Neuroinvasion typically requires efficient viral transmission through a polarized epithelium followed by long-distance transport through the viscous axoplasm. These events are mediated by the recruitment of the cellular microtubule motor proteins to the intracellular viral particle and by alterations to the cytoskeletal architecture. The focus of this review is the interplay between neurotropic herpesviruses and the cytoskeleton.

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Section: Alphaherpesviruses and The Host Cell Cytoskeleton: Interactimentioning

confidence: 99%

Alphaherpesviruses and the Cytoskeleton in Neuronal Infections

Zaichick

Bohannon

Smith

2011

Viruses

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“…Recently, more research has focused on PHEV because infection rates have risen in some countries (Cartwright et al, 1969;Gao et al, 2011;Mengeling and Cutlip, 1976;Quiroga et al, 2008;Rho et al, 2011). This virus spreads to the CNS via peripheral nerves, and nerve cells are one target for viral replication (Hara et al, 2009). In addition, PHEV can be isolated from primary porcine kidney cell cultures (Hirano et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

Porcine hemagglutinating encephalomyelitis virus induces apoptosis in a porcine kidney cell line via caspase-dependent pathways

et al. 2013

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Porcine hemagglutinating encephalomyelitis is an acute, highly contagious disease in piglets that is caused by the porcine hemagglutinating encephalomyelitis virus (PHEV). However, the pathogenesis of PHEV and the relationship between PHEV and the host cells are not fully understood. In this study, we investigated whether the PHEV-induced cytopathic effect (CPE) was caused by apoptosis. Replication of PHEV in a porcine kidney-derived cell line (PK-15 cells) caused an extensive CPE, leading to the destruction of the entire monolayer and the death of the infected cells. Staining with Hoechst 33,342 revealed morphological changes in the nuclei and chromatin fragmentation. In addition, PHEV caused DNA fragmentation detectable by agarose gel electrophoresis 48h post-infection, increasing with the incubation time. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 96h post-infection, as determined using flow cytometry and fluorescence microscopy of cells that were stained with annexin V-FITC and propidium iodide (PI). Moreover, as is commonly observed for coronavirus infections of other animals, the activities of the effecter caspase, caspase-3, and the initiator caspases, caspase-8 and caspase-9, which are representative factors in the death receptor-mediated apoptotic pathway and the mitochondrial apoptotic pathway, respectively, were increased in PHEV-infected PK-15 cells. Moreover, the tripeptide pan-ICE (caspase) inhibitor Z-VAD-FMK blocked PHEV-induced apoptosis but did not have an effect on virus production by 96h post-infection. These results suggested that PHEV induces apoptosis in PK-15 cells via a caspase-dependent pathway. Apoptotic death of infected cells is detrimental to animals because it causes cell and tissue destruction. Although the pathological characteristics of PHEV are largely unknown, apoptosis may be the pathological basis of the lesions resulting from PHEV infection.

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“…Even if an infected pig survives the diseaseassociated immunopathology or the toxicity of the drug treatment, PHE can acutely affect its growth. Increasing research attention has been focused on PHE because infection rates have increased in some countries [31][32][33][34] . PHE is caused by PHE-CoV, a virus that has only one known strain and is the only known CoV that is neurotropic in pigs [25,35,36] .…”

Section: Discussionmentioning

confidence: 99%

In vitro Inhibition of Porcine Hemagglutinating Encephalomyelitis Virus Replication with siRNAs Targeting the Spike Glycoprotein and Replicase Polyprotein Genes

Lan

Zhao

et al. 2011

Intervirology

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Objective: The specific effect of RNA interference on the replication of porcine hemagglutinating encephalomyelitis virus (PHE-CoV) was explored. Methods: Four species of small interfering RNA (siRNA), targeting different regions of the PHE-CoV spike glycoprotein and replicase polyprotein genes, were prepared by in vitro transcription. After transfection of PK-15 cells with each of the siRNAs followed by infection with PHE-CoV, the cytopathic effect (CPE) was examined by phase-contrast microscope, and viral proliferation within cells was examined by indirect immunofluorescence microscopy, hemagglutination (HA) test, TCID₅₀ assay and real-time RT-PCR. Results: Examination of CPE demonstrated that the four siRNAs were capable of protecting cells against PHE-CoV invasion with very high specificity and efficiency. At 48 h post-infection, only a few siRNA-treated cells were positive for viral antigen staining, whereas most untreated virus-infected cells were positive. Transfection with siRNAs also suppressed the production of infectious virus by up to 18- to 32-fold as assessed by a HA test and 93- to 494-fold as assessed by TCID₅₀ assay. Furthermore, treatment with siRNAs caused a 53–91% reduction in the viral genome copy number as assessed by real-time RT-PCR. Conclusion: These results suggested that the four species of siRNAs can efficiently inhibit PHE-CoV genome replication and infectious virus production.

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Propagation of Swine Hemagglutinating Encephalomyelitis Virus and Pseudorabies Virus in Dorsal Root Ganglia Cells

Cited by 22 publications

References 28 publications

Alphaherpesviruses and the Cytoskeleton in Neuronal Infections

Alphaherpesviruses and the Cytoskeleton in Neuronal Infections

Porcine hemagglutinating encephalomyelitis virus induces apoptosis in a porcine kidney cell line via caspase-dependent pathways

In vitro Inhibition of Porcine Hemagglutinating Encephalomyelitis Virus Replication with siRNAs Targeting the Spike Glycoprotein and Replicase Polyprotein Genes

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