An Improved Method for Extracting Cell-Free Herpesviruses of Marek's Disease and Turkeys from Infected Cell Cultures

Br, Cho

doi:10.2307/1589522

Cited by 8 publications

(5 citation statements)

References 14 publications

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“…Table 5 shows the titers of celt-free viruses obtained from infected CEF by sonic extraction with various suspending media. In general, cell-free virus was extracted more easily with SPGA and SPGA with 10 per cent sorbitol than PBS and MEM with 10 per cent FCS as reported by others (7,9). The titers of cell-free type 2 PPA extracted with the four different media were similar to those of FC-126.…”

Section: Biological Characterization O/type 2 P Pasupporting

confidence: 71%

“…Eagle's minimum essential medium (MEM, Nissui, Japan) supplemented with 10 per cent FCS; 3. sucrose-phosphateg]utaminate-albumin buffer (SPGA) (7) ; and 4. SPGA supplemented with 10 per cent sorbitel (9). Harvested cells were washed once with the above suspending media, extracted sonically (18) and then centrifuged at 6,000×g for 20 minutes.…”

Section: ~ I~i2isawa Et Almentioning

confidence: 99%

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A comparison of the biological properties of type 2 plaque-producing agent derived from the Cal-1 strain of Marek's disease virus with other related viruses

Kirisawa

Hirai

Yachida

et al. 1986

Archives of Virology

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The serological and biological properties of the type 2 plaque-producing agent (PPA) derived from the Cal-1 strain of Marek's disease virus (MDV) were compared with those of reference strains of the three serotypes of MDV and herpesvirus of turkeys (HVT) groups; namely JM, HPRS-24 strains of MDV and the FC-126 strain of HVT for serotype 1, 2, and 3, respectively. By agar gel precipitation (AGP), indirect fluorescent antibody and virus neutralization tests, type 2 PPA was related but not identical to the FC-126 strain. By the AGP test, type 2 PPA showed a poor ability to synthesize B antigen and the A antigen was different from that of strain FC-126. To compare the virological characteristics of type 2 PPA with the reference strains, the release of cell-free virus into supernatants of infected cell cultures and titers of cell-free virus extracted sonically from infected cell cultures were examined. Cell-free type 2 PPA virus was easily detected in the supernatants and extracted from infected cell cultures. These properties were similar to reference strains of serotype 2 and 3. Next, the structural similarities of viral DNAs of type 2 PPA and strain FC-126 were examined by Southern blot hybridization. The restriction endonuclease-cleavage patterns of DNA of type 2 PPA were very similar but not identical to those of the FC-126 strain. In chickens inoculated with type 2 PPA, splenic lymphocytes supported a non-productive latent infection as did also those from chickens inoculated with the FC-126 or HPRS-24 strains. From these results, type 2 PPA appears to belong to serotype 3 of MDV and HVT groups. The origin of type 2 PPA is discussed.

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Section: Biological Characterization O/type 2 P Pasupporting

confidence: 71%

Section: ~ I~i2isawa Et Almentioning

confidence: 99%

A comparison of the biological properties of type 2 plaque-producing agent derived from the Cal-1 strain of Marek's disease virus with other related viruses

Kirisawa

Hirai

Yachida

et al. 1986

Archives of Virology

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“…Lack of knowledge of receptors for the MDV entry, which helps to determine host range and tissue tropism, will certainly limit our understanding of the cascade of events in the early pathogenic processes. Various extraction methods have been described for cell-free MDVs from infected cell cultures, but were reported to be successful only for in vitro infection [3,5,7,9,12,23]. Here, we reported that cell-free MDV prepared in this study was high titer of infectious virus, and useful to study the infection mechanism in vivo as well as in vitro.…”

Section: Discussionmentioning

confidence: 74%

“…Briefly, infected cultures after incubation for 5 to 7 days were drained and rinsed once with phosphatebuffered saline (PBS). The cultures were treated with trypsin-versene solution, transferred to tubes, washed with SPGA-SBT (0.4 mM KH 2 PO 4 , 7.2 mM K 2 HPO 4 , 7.46% sucrose, 1% BSA, 10% sorbitol and 0.92 mg/ml sodium glutamate), and disrupted in SPGA-SBT by sonication (Astrason Ultrasonic Processor, Misonix Inc., NY) at a 50% probe intensity (20 kHz) for 1 min [9]. Then, the sonicated solutions were centrifuged at 2,000 × g for 5 min to remove the cell debris, and the supernatants were aliquoted and stored at -80°C.…”

Section: Methodsmentioning

confidence: 99%

Heparin Inhibits Plaque Formation by Cell-free Marek's Disease Viruses in vitro.

Lee

Ohashi

Sugimoto

et al. 2001

The Journal of Veterinary Medical Science

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ABSTRACT. The mechanisms of Marek's disease virus (MDV) entry to host cells have not yet been analyzed. Heparan sulfate (HS) on the cell surface serves as a receptor for several herpesviruses in mammalian species. In this study, we demonstrated that plaque formation by cell-free MDV is inhibited by the addition of soluble heparin to the cell culture. Moreover, pretreatment of susceptible cells, chicken embryo fibroblasts, with heparinase, partially reduced infectivity of the cell-free MDV. From these results, it was suggested that the MDV entry, at least in the case of cell-free MDV, is dependent on the presence of cell surface glycosaminoglycans, principally HS. KEY WORDS: cell-free MDV, heparan sulfate, heparin.J. Vet. Med. Sci. 63(4): 427-432, 2001 The primary event of any viral infection is attachment of virus to its host cell. Viral receptors are often involved in defining the host range and specific tissue tropism of a virus. Attachment of herpesviruses to target cells is mediated by viral glycoproteins which are embedded in the virion envelope, and which interact with cellular surface components acting as virus receptors. The first virus found to bind to heparan sulfate (HS) was herpes simplex virus (HSV) [43], and since then, a number of herpesviruses have been demonstrated to use HS as an initial receptor for their entry. In the cases of the alphaherpesviruses, initial interaction between the virion and the target cell involves binding of glycoprotein C (gC) [37] and/or glycoprotein B (gB) [22] to cell-surface glycosaminoglycans (GAG), in particular, HS, which are components of proteoglycans. This interaction with HS has been observed for HSV-1, and HSV-2 [43], pseudorabies virus (PRV) [24], varicella-zoster virus [44], and bovine herpesvirus 1 (BHV-1) [27]. In addition, the gamma- [40] and betaherpesviruses [10,35] Marek's disease virus (MDV), an avian herpesvirus, is the causative agent of Marek's disease (MD), which is characterized by CD4 + T cell lymphoma and nerve enlargement caused by infiltration of lymphocytes and/or lymphoma cells. The pathogenesis of MD can be sequentially divided into three phases: early cytolytic infection; latent infection; and secondary cytolytic infection with immunosuppression and tumor development [26,32,34]. Early cytolytic infection occurs mainly in B cells, and latently infected lymphocytes are mainly activated T cells [36]. In addition, target cells for transformation by MDV are mainly CD4 + T cells [34], suggesting that latent infection in the T cell subsets could be intimately related to the subsequent transformation by MDV. MDVs are highly cell-associated, and the cellfree virus particles are produced only in feather follicle epithelium (FFE) of MDV-infected chickens [6]. However, the mechanism(s) by which MDV attaches to its host cells has not been delineated.Here, we demonstrated that, cell-free MDVs, experimentally prepared from cell culture in vitro, were infectious to cells not only in vitro but also in vivo. In addition, we also showed that cell surfac...

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“…Genetic heterogeneity in a herpes simplex virus type-1 clinical isolate was also reported, and this genetic heterogeneity was associated with viral pathogenicity (Montgomery & Centifanto, 1989). Although it is possible to obtain MDV populations from one single virion that is artificially extracted from infected cells (Cho, 1978), the extraction is extremely inefficient, especially with virulent strains.…”

Section: Introductionmentioning

confidence: 99%

Virulent Marek's disease virus generated from infectious bacterial artificial chromosome clones with complete DNA sequence and the implication of viral genetic homogeneity in pathogenesis

Niikura

Kim

Silva

et al. 2010

Journal of General Virology

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An Improved Method for Extracting Cell-Free Herpesviruses of Marek's Disease and Turkeys from Infected Cell Cultures

Cited by 8 publications

References 14 publications

A comparison of the biological properties of type 2 plaque-producing agent derived from the Cal-1 strain of Marek's disease virus with other related viruses

A comparison of the biological properties of type 2 plaque-producing agent derived from the Cal-1 strain of Marek's disease virus with other related viruses

Heparin Inhibits Plaque Formation by Cell-free Marek's Disease Viruses in vitro.

Virulent Marek's disease virus generated from infectious bacterial artificial chromosome clones with complete DNA sequence and the implication of viral genetic homogeneity in pathogenesis

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