Isolation, characterization, and physical mapping of a pseudorabies virus mutant containing antigenically altered gp50

Wathen, Michael W.; Wathen, L. M. K.

doi:10.1128/jvi.51.1.57-62.1984

Cited by 81 publications

(42 citation statements)

References 43 publications

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“…As controls, anti-gII(PrV) monoclonal antibody (MAb) 5/14 (kindly provided by H.-J. Rziha, Tubingen, Germany [17]) and anti-gpSO MAb MCA50-1 (kindly provided by M. W. Wathen, Kalamazoo, Mich. [36]) were used. Precipitates were separated in nonreducing sodium dodecyl sulfate-10% polyacrylamide gels VIRULENCE OF gp50-PrV Mice were infected intranasally with different PFU of gX-Pgal (gX-) mutants derived from either wild-type strain NIA-3 or Ka.…”

Section: Methodsmentioning

confidence: 99%

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Glycoprotein gp50-negative pseudorabies virus: a novel approach toward a nonspreading live herpesvirus vaccine

Heffner¹,

Kovács²,

Klupp³

et al. 1993

J Virol

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Essential herpesvirus glycoproteins are involved in membrane fusion processes during infection, e.g., viral penetration and direct cell-to-cell transmission. We previously showed that the gD-homologous glycoprotein gpSO of pseudorabies virus (PrV) is essential for virus entry into target cells but proved to be dispensable for direct viral cell-to-cell spread in cell culture (I. Rauh and T. C. Mettenleiter, J. Virol. 65:5348-5456, 1991). For gp5O-negative (gp50-) viruses, after phenotypic complementation necessary for primary infection, the only means of viral spread is by way of direct cell-to-cell transmission. In contrast, virus mutants lacking the essential gB-homologous glycoprotein gll after phenotypic complementation are only able to infect primary target cells and are blocked in further viral spread. To analyze how these in vitro phenotypes translate into virus replication in the animal, mice were infected intranasally with gp50or gII-PrV mutants after prior phenotypic complementation by propagation on cell lines providing the essential glycoprotein in trans. Our results show that whereas the gIImutants did not cause disease or any symptoms, gp50mutants derived from two different PrV strains were fully virulent, with animals exhibiting severe symptoms ultimately leading to death. However, free infectious virus could not be recovered from either gp50or gIl-PrV-infected animals. We conclude that direct cell-to-cell transmission as the only means of viral spread of the gp50-* Corresponding author.

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Section: Methodsmentioning

confidence: 99%

“…Glycoprotein gpSO has been shown to represent a major immunogenic protein of PrV, and several potent neutralizing anti-gp5O antibodies have been described (3,8,36). In addition, vaccination with recombinant gpSO has been shown to protect animals from AD (18,30).…”

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confidence: 99%

Glycoprotein gp50-negative pseudorabies virus: a novel approach toward a nonspreading live herpesvirus vaccine

Heffner¹,

Kovács²,

Klupp³

et al. 1993

J Virol

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“…While considerable progress has been made in the identification and localization of glycoprotein genes for a variety of herpesviruses (3,6,9,11,13,15,17,20,22,25,31,33,35), the functional and genetic analysis of glycoprotein gene expression has advanced more slowly. One reason is that until recently, few good selection or screening techniques existed for the direct analysis of glycoproteins.…”

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confidence: 99%

Pseudorabies virus gene encoding glycoprotein gIII is not essential for growth in tissue culture

Robbins¹,

Whealy²,

Watson³

et al. 1986

J Virol

101

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We have established that in the Becker strain of pseudorabies virus (PRV), the glycoprotein glll gene is not essential for growth in cell culture. This was accomplished by construction and analysis of viral mutants containing two defined deletion mutations affecting the glll gene. These mutations were first constructed in vitro and introduced into Escherichia coli expression plasmids to verify structure and protein production. Each mutation was then crossed onto PRV by cotransfection of plasmid DNA and parental viral DNA by using glll-specific monoclonal antibodies as selective and screening reagents. One resultant virus strain, PRV-2, contained an in-frame deletion of a 402-base-pair (bp) Sacl fragment contained within the glll gene. Another virus strain, PRV-10, contained a deletion of a 1,480-bp XhoI fragment removing 230 bp of the upstream, putative transcriptional control sequences and 87% of the gIll coding sequence. The deletion mutants were compared with parental virus by analysis of virion DNA, glll specific RNA, and proteins reacting with glll specific antibodies. Upon infection of PK15 cells, the deletion mutants did not produce any proteins that reacted with two glll specific monoclonal antibodies. However, two species of truncated glycosylated proteins were observed in PRV-2 infected cells that reacted with antiserum raised against bacterially produced gIll protein. PRV-10 produced no detectable gIll-specific RNA or protein. PRV-10 could be propagated without difficulty in tissue culture. Virus particles lacking gIII were indistinguishable from parental PRV virus particles by analysis of infected-cell thin sections in the electron microscope. We therefore conclude that expression of the gIll gene was not absolutely essential for PRV growth in tissue culture. 635on August 1, 2020 by guest

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“…Glycoproteins gB, gD, gH and gL are known to be essential for viral replication and although gC, gE, gG, and gl can be deleted, the fact that no field isolates have been found which lack these glycoproteins suggests an important role in the biology of the virus in vivo. Genes for glycoproteins gD, gE, gG and gl are found in the unique short segment of the genome while gB, gC, gH, gK, gL, gM and gN are found in the unique long segment [110,130,131,181,184,220,230,236,285].…”

Section: Glycoproteinsmentioning

confidence: 99%

Evaluation of vaccinia vectored vaccines expressing glycoproteins gB, gC or gD of pseudorabies virus

Brockmeier

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This study evaluated vaccinia-vectored vaccines expressing glycoproteins gB, gC, or gD of pseudorabies virus (PRV) in pigs. The vaccines are based on an attenuated vaccinia virus strain, NYVAC, which has reduced virulence and replicative capacity for certain species, including swine. The recombinant vaccinia vaccines were unable to prevent replication of virulent pseudorabies virus or latency but they were able to decrease the amount of virus shed after challenge and decrease clinical signs. In particular, pigs vaccinated with the recombinants expressing either gB or gD were protected at a level comparable to an inactivated PRV virus that was given for comparison. No lesions or clinical signs were seen following vaccination in these studies and no seronegative pigs in contact with vaccinia-vaccinated pigs ever seroconverted. No significant increase in protection occured by giving recombinants expressing multiple glycoproteins and there was no virus neutralizing antibody response or protection induced when the recombinant vaccines were given orally or intranasally. The response to the recombinants is dose dependent but there was no indication that prior immunity to the vaccinia parent virus had any inhibitory effect on subsequent vaccination with these recombinants. The gB recombinant, but not the gD recombinant, was able to stimulate an active immune response in piglets with passive immunity fix)m PRV or NYVAC/gD vaccinated dams. The NYVACyPRV recombinant vaccines were compatible with differential serologic tests currently on the market We could not demonstrate significant cell-mediated immune responses after vaccination with the recombinant vaccines but they may prime this type of response. In summary, the NYVAC/PRV recombinant vaccines afforded protection comparable with that of PRV vaccines currentiy on the market, were safe, and offer some advantages over currentiy available vaccines such as elimination of attenuated PRV viral strains, better induction of immunity in the presence of passive immunity, and more sensitive differential testing capabilities.

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Isolation, characterization, and physical mapping of a pseudorabies virus mutant containing antigenically altered gp50

Cited by 81 publications

References 43 publications

Glycoprotein gp50-negative pseudorabies virus: a novel approach toward a nonspreading live herpesvirus vaccine

Glycoprotein gp50-negative pseudorabies virus: a novel approach toward a nonspreading live herpesvirus vaccine

Pseudorabies virus gene encoding glycoprotein gIII is not essential for growth in tissue culture

Evaluation of vaccinia vectored vaccines expressing glycoproteins gB, gC or gD of pseudorabies virus

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