Low Infectivity of HSV-1 DNA Caused by Defective-interfering Genomes

Hennes-Stegmann, B.; Schröder, Claus H.

doi:10.1099/0022-1317-63-2-307

Cited by 8 publications

(2 citation statements)

References 17 publications

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“…Interference by DI particles has been described in various virus systems including vesicular stomatitis virus (18), rotavirus (27), avian leukosis virus (39), herpes simplex (14) and pseudorabies viruses (3), and the parvoviruses, adeno-associated virus (AAV) (7), H-I (38), and Lu III (33). Empty (noninfectious) parvovirus particles have also been suggested as an additional viral interference factor in parvovirus replication (23,42).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of porcine parvovirus replication by empty virus particles

Choi

Molitor

Joo

1987

Archives of Virology

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The influence of empty porcine parvovirus (PPV) particles on viral replication was examined in cell cultures and in swine. Following extensive purification, homogeneous preparations of full and empty PPV preparations were obtained and used for in vitro and in vivo analyses. In the first in vitro experiment, swine testes cells were infected with mixtures of various ratios of empty and full (E/F) particles. The production of both intracellular and extracellular virus was markedly inhibited in the presence of empty particles. This inhibition was dependent upon the concentration of empty particles present in the mixture. In the second in vitro study, various concentrations of empty particles were added prior to full virus infection. Again, marked inhibition of progeny virus production was evident and related to the concentrations of empty particles added. Based on the results of in vitro studies, the influence of empty particles on PPV infection in swine was tested by infecting mid-term and late-term gestation swine fetuses with various E/F particle ratios. Both mid-term and late-term fetuses exposed to 0:1, 1:1 and 5:1 E/F ratios displayed gross pathological evidence of PPV infection whereas fetuses exposed to E/F ratios of 30:1 or greater were grossly normal in appearance. However, fetuses infected with 30:1, 50:1 and 300:1 E/F ratios showed evidence of virus in their tissues by DNA hybridization. Regardless of the E/F ratios, late-term infected fetuses responded with high antibody titers ranging from 1024 to 4096. The results from these studies suggested that empty particles interfered with viral replication in both cell culture and in animals.

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Section: Introductionmentioning

confidence: 99%

Inhibition of porcine parvovirus replication by empty virus particles

Choi

Molitor

Joo

1987

Archives of Virology

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“…Therefore, increased expression of ICP4 presumably not only led to higher levels of early and late classes of genes but may serve as a protection function in the stability of the viral genome as evidenced by enhancement in total HSV-1 genome copy number. The greater differential in total genome copy number can be reflected in the generation of defective interfering viral genomic DNA that occurs during serial passaging of HSV-1 (Hennes-Stegmann and Schroder, 1982;Schroder et al, 1984).…”

Section: Discussionmentioning

confidence: 99%

Engineering cell lines for production of replication defective HSV‐1 gene therapy vectors

Grant

Krisky²,

Ataai

2008

Biotech & Bioengineering

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Herpes simplex virus type 1 (HSV-1) represents an attractive vehicle for a variety of gene therapy applications. To render this virus safe for clinical use, its cytotoxic genes must be removed without losing its ability to express transgenes efficiently. Our vectors are deleted for the essential immediate early genes ICP4 and ICP27. These genes are controlled by unique promoters having enhancer elements responsive to a viral structural protein VP16. The expression of these genes occurs prior to the activation of all other lytic functions and is thus required to initiate and complete the virus replication cycle. For large scale manufacture of clinical grade vectors, efficient cell lines must be generated that express the essential viral gene products in trans during vector propagation. Here we describe methods for engineering HSV-1 production cell lines that improve vector growth by altering the kinetics of complementing gene expression. We examined the ability of Vero cells independently transduced with ICP4 and ICP27 under transcriptional control of their respective promoters to support the growth of a replication defective vector (JDTOZHE), deleted for ICP4, ICP27 and approximately 20 kb of internal elements that are not required for virus growth in Vero cells. Vector yield on this cell line was 3 logs lower than wild-type virus grown on Vero cells. To understand the mechanism underlying poor vector yield, we examined the expression of ICP4 and ICP27 during virus complementation. While ICP27 was expressed immediately on vector infection, the expression of ICP4 was considerably delayed by 8-10 h, suggesting that the ICP4 promoter was not adequately activated by VP16 delivered by the infectious vector particle. Use of the ICP0 promoter to express ICP4 from the cellular genome resulted in higher induction levels and faster kinetics of ICP4 expression and a 10-fold improvement in vector yield. This study suggests that vector complementation is highly dependent on the kinetics of complementing gene expression and can lead to large differences in vector yield.

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Cyclosporin a resistance of Herpes simplex virus-induced “fusion from within” as a phenotypical marker of mutations in the syn 3 locus of the glycoprotein B gene

et al. 1994

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We here report research in which nine strains of Herpes simplex virus (HSV) with fusing activity were investigated in order to establish precise phenotypical markers of mutations in the carboxy terminus of glycoprotein B (gB). The gene region encoding the carboxy terminus of gB was isolated, then cloned, and finally sequenced. Our investigation showed that seven strains have different mutations in the syn 3 locus. We observed no base difference in the gB gene region encoding the carboxy terminus of gB of two other strains. Strains with a mutation in the carboxy terminus of gB induced fusion from within (FFWI) in the presence of Cyclosporin A (CyA) at a concentration up to 150 microM. There are two clusters of mutations correlated with the syn 3 locus and selected in the presence of CyA: One group comprised of amino acid substitutions at position 816, the other of changes at positions 853, 854, and 857. In contrast, the fusion induced by strains with mutations in other syn loci is CyA sensitive. CyA inhibits the FFWI at concentrations of 20-60 microM. The results demonstrate the CyA resistance of HSV-induced FFWI should serve as a phenotypical marker of mutations in the carboxy terminus of gB. Moreover, our investigations revealed that fusion from without (FFWO) does not always serve as a phenotypical marker of mutations in the syn 3 locus. On the one hand, all FFWO-positive strains possess a syn 3 locus mutation, whilst, on the other hand, five strains with mutations in the carboxy terminus of gB are FFWO negative.(ABSTRACT TRUNCATED AT 250 WORDS)

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Low Infectivity of HSV-1 DNA Caused by Defective-interfering Genomes

Cited by 8 publications

References 17 publications

Inhibition of porcine parvovirus replication by empty virus particles

Inhibition of porcine parvovirus replication by empty virus particles

Engineering cell lines for production of replication defective HSV‐1 gene therapy vectors

Cyclosporin a resistance of Herpes simplex virus-induced “fusion from within” as a phenotypical marker of mutations in the syn 3 locus of the glycoprotein B gene

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