A Comparative Study of Pseudorabies Virus (PRV) Strains with Defects in Thymidine Kinase and Glycoprotein Genes

Ferrari, M; Mettenleiter, Thomas C.; Romanelli, Maria Grazia; Cabassi, E.; Corradi, A.; Mas, Nora; Silini, R.

doi:10.1053/jcpa.2000.0406

Cited by 24 publications

(31 citation statements)

References 18 publications

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“…The enzyme thymidine kinase (TK) of alphaherpesviruses is a product of an early gene required for viral DNA replication in neurons, but is not essential for viral replication in epithelial cells or in cell cultures (Chen et al 1998, 2004, Enquist et al 1998. Even though TK is non-essential for viral replication in dividing cells, alphaherpesviruses defective in TK activity usually show a reduced ability to replicate in peripheral tissues compared to their TK+ counterparts (Kaashoek et al 1996, Ferrari et al 2000. Herein, we demonstrated that BoHV-5TKΔ replicates with reduced efficiency in the nasal mucosa of rabbits.…”

Section: Discussionmentioning

confidence: 99%

“…Nonetheless, TK is required for the full expression of herpesvirus neurovirulence in vivo (Enquist et al 1998). Deletion of TK gene in HSV-1 and PRV has been associated with deficient replication in neurons and reduced neurovirulence (Chen et al 2004, Ferrari et al 2000. The ability to establish latent infection does not seem to be affected in TK-negative viruses, like HSV-1, PRV and bovine herpesvirus type 1 (BoHV-1) (Chen et al 2004, Ferrari et al 2000, Whetstone et al 1992.…”

Section: Introductionmentioning

confidence: 99%

“…Deletion of TK gene in HSV-1 and PRV has been associated with deficient replication in neurons and reduced neurovirulence (Chen et al 2004, Ferrari et al 2000. The ability to establish latent infection does not seem to be affected in TK-negative viruses, like HSV-1, PRV and bovine herpesvirus type 1 (BoHV-1) (Chen et al 2004, Ferrari et al 2000, Whetstone et al 1992. However, the ability to reactivate the infection is usually severely impaired, if not abolished, depending on the viruses, animal and experimental procedures (Coen et al 1989, Mengeling 1991, Whetstone et al 1992, Ferrari et al 2000.…”

Section: Introductionmentioning

confidence: 99%

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A thymidine kinase-negative bovine herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection

et al. 2011

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ABSTRACT.-Silva S.C., Brum M.C.S., Oliveira S.A.M., Weiblen R. & Flores E.F. 2011. A thymidine kinase-negative bovine herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection. Pesquisa Veterinária Brasileira 31(5):389-397. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Camobi, Santa Maria, Mutant viral strains deleted in non-essential genes represent useful tools to study the function of specific gene products in the biology of the virus. We herein describe an investigation on the phenotype of a bovine herpesvirus 5 (BoHV-5) recombinant deleted in the gene encoding the enzyme thymidine kinase (TK) in rabbits, with special emphasis to neuroinvasiveness and the ability to establish and reactivate latent infection. Rabbits inoculated with the parental virus (SV-507/99) (n=18) at a low titer (10 TCID 50 ) and for a shorter period (average: 6.6 days [2-11]) and remained healthy. PCR examination of brain sections of inoculated rabbits at day 6 post-infection (pi) revealed a widespread distribution of the parental virus, whereas DNA of the recombinant BoHV-5TKΔ-was detected only in the trigeminal ganglia [TG] and olfactory bulbs [OB]. Nevertheless, during latent infection (52pi), DNA of the recombinant virus was detected in the TGs, OBs and also in other areas of the brain, demonstrating the ability of the virus to invade the brain. Dexamethasone (Dx) administration at day 65 pi was followed by virus reactivation and shedding by 5/8 rabbits inoculated with the parental strain (mean duration of 4.2 days [1 -9]) and by none of seven rabbits inoculated with the recombinant virus. Again, PCR examination at day 30 post-Dx treatment revealed the presence of latent DNA in the TGs, OBs and in other areas of the brain of both groups. Taken together, these results confirm that the recombinant BoHV-5TKΔ is highly attenuated for rabbits. It shows a reduced ability to replicate in the nose but retains the ability to invade the brain and to establish latent infection. Additional studies are underway to determine the biological and molecular mechanisms underlying the inability of BoHV-5TKΔ to reactivate from latency.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A thymidine kinase-negative bovine herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection

et al. 2011

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“…We therefore sought to employ a replication-defective PrV strain to construct a single helper virus to provide replication and packaging functions for rAAV vector production. Since the TK activity of herpesviruses has been associated with virulence, inactivation of the TK gene in the PrV genome attenuates the virus (14). With regard to safety concerns for gene therapy, recombinant viral vectors should not spread out to be transmitted to other uninoculated individuals.…”

Section: Vol 79 2005mentioning

confidence: 99%

“…Moreover, PrV-encoded thymidine kinase (TK) serves as another virulence factor to support viral DNA replication. PrV mutants that are defective in TK function reduce virulence, replication in peripheral target tissues, and migration to the central nervous system (14). For this study, we constructed a gD/gE/TK triply defective recombinant PrV, designated PS virus, to express the AAV Rep and Cap proteins and then employed it as a helper virus accompanied with packaging machinery for the production of rAAV vectors.…”

mentioning

confidence: 99%

Novel Strategy for Generation and Titration of Recombinant Adeno-Associated Virus Vectors

Shiau

Liu

2005

J Virol

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Recombinant adeno-associated virus (rAAV) vectors have many advantages for gene therapeutic applications compared with other vector systems. Several methods that use plasmids or helper viruses have been reported for the generation of rAAV vectors. Unfortunately, the preparation of large-scale rAAV stocks is labor-intensive. Moreover, the biological titration of rAAV is still difficult, which may limit its preclinical and clinical applications. For this study, we developed a novel strategy to generate and biologically titrate rAAV vectors. A recombinant pseudorabies virus (PrV) with defects in its gD, gE, and thymidine kinase genes was engineered to express the AAV rep and cap genes, yielding PS virus, which served as a packaging and helper virus for the generation of rAAV vectors. PS virus was useful not only for generating high-titer rAAV vectors by cotransfection with an rAAV vector plasmid, but also for amplifying rAAV stocks. Notably, the biological titration of rAAV vectors was also feasible when cells were coinfected with rAAV and PS virus. Based on this strategy, we produced an rAAV that expresses prothymosin ␣ (ProT). Expression of the ProT protein in vitro and in vivo mediated by rAAV/ProT gene transfer was detected by immunohistochemistry and a bioassay. Taken together, our results demonstrate that the PrV vector-based system is useful for generating rAAV vectors carrying various transgenes.

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Comparison of immune responses to different foot-and-mouth disease genetically engineered vaccines in guinea pigs

Yao

Qian

Huang

et al. 2008

Journal of Virological Methods

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A Comparative Study of Pseudorabies Virus (PRV) Strains with Defects in Thymidine Kinase and Glycoprotein Genes

Cited by 24 publications

References 18 publications

A thymidine kinase-negative bovine herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection

A thymidine kinase-negative bovine herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection

Novel Strategy for Generation and Titration of Recombinant Adeno-Associated Virus Vectors

Comparison of immune responses to different foot-and-mouth disease genetically engineered vaccines in guinea pigs

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