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Journal of virology1999; 73(9); 7399-7409; doi: 10.1128/JVI.73.9.7399-7409.1999

Equine herpesvirus 1 gene 12 can substitute for vmw65 in the growth of herpes simplex virus (HSV) type 1, allowing the generation of optimized cell lines for the propagation of HSV vectors with multiple immediate-early gene defects.

Abstract: Herpes simplex virus (HSV) has often been suggested for development as a vector, particularly for the nervous system. Considerable evidence has shown that for use of HSV as a vector, immediate-early (IE) gene expression must be minimized or abolished, otherwise such vectors are likely to be highly cytotoxic. Mutations of vmw65 which abolish IE promoter transactivating activity may also be included to reduce IE gene expression generally. However, when vmw65 mutations are combined with an IE gene deletion, such viruses are hard to propagate, even on cells which otherwise complement the IE gene deletion effectively. We have found that vmw65 mutants can be effectively grown on cell lines expressing equine herpesvirus 1 gene 12, a non-HSV homologue of vmw65 with little sequence similarity to its HSV counterpart. This prevents repair by homologous recombination of vmw65 mutations in the virus, which would occur if mutations were complemented by vmw65 itself. The gene 12 protein is not packaged into HSV virions, which is important if viruses grown on such cells are to be used as vectors. These results not only further strengthen the evidence for direct functional homology between and similar modes of action of the two proteins but have allowed the generation of gene 12-containing cell lines in which ICP4 and ICP27 expression is induced by virus infection (probably by ICP0) and which give efficient growth of viruses deficient in ICP27, ICP4, and vmw65 (the viruses also have ICP34.5/ORFP deleted). Efficient growth of such viruses has not previously been possible. As these viruses are highly deficient in IE gene expression generally, such virus-cell line combinations may provide an alternative to HSV vectors with deletions of all four of the regulatory IE genes which, for optimal growth, require cell lines containing all four IE genes but which are hard to generate due to the intrinsic cytotoxicity of each of the proteins.
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Publication Date: 1999-08-10 PubMed ID: 10438830PubMed Central: PMC104267DOI: 10.1128/JVI.73.9.7399-7409.1999Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research paper discusses the development and propagation of a safer vector for the herpes simplex virus (HSV) type 1 by substituting a specific viral gene, vmw65, with an analogous gene from equine herpesvirus 1, known as gene 12. This substitution allows the creation of optimized cell lines that diminish the cytotoxicity caused by immediate-early gene expression in HSV vector-based treatments.

Motivation for the Research

  • HSV is frequently proposed for use as a vector in medical treatments, particularly those involving the nervous system.
  • However, the expression of immediate-early (IE) genes in HSV makes these vectors highly cytotoxic, limiting their applications.
  • To reduce this cytotoxicity, the researchers aim to minimize or eliminate IE gene expression.
  • A typical strategy includes mutations in vmw65, a gene which regulates IE gene expression, but this approach often results in HSV vectors that are difficult to grow and propagate.

The Role of Equine Herpesvirus 1 Gene 12

  • The authors discovered that vmw65 mutant HSV can be efficiently grown on cell lines expressing equine herpesvirus 1 gene 12, an analogous but not identical gene to vmw65 from a different virus species.
  • The use of gene 12 prevents the repair of vmw65 mutations in HSV through homologous recombination, which could restore its cytotoxicity.
  • Importantly, the gene 12 protein is not incorporated into HSV virions, making it a safe choice for the development of vectors.

Outcomes and Implications of the Research

  • The use of equine herpesvirus 1 gene 12 enabled the creation of cell lines capable of efficient growth of HSV deficient in ICP27, ICP4, and vmw65; an outcome previously unattainable.
  • The resulting HSV vectors exhibit reduced IE gene expression, mitigating the typical cytotoxicity associated with HSV vectors.
  • This research provides a novel alternative to prior strategies for creating less cytotoxic HSV vector, and could lead to the development of safer, more effective HSV-based treatments.

Cite This Article

APA
Thomas SK, Lilley CE, Latchman DS, Coffin RS. (1999). Equine herpesvirus 1 gene 12 can substitute for vmw65 in the growth of herpes simplex virus (HSV) type 1, allowing the generation of optimized cell lines for the propagation of HSV vectors with multiple immediate-early gene defects. J Virol, 73(9), 7399-7409. https://doi.org/10.1128/JVI.73.9.7399-7409.1999

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 73
Issue: 9
Pages: 7399-7409

Researcher Affiliations

Thomas, S K
  • Department of Molecular Pathology, The Windeyer Institute of Medical Sciences, University College London, London W1P 6DB, England, UK.
Lilley, C E
    Latchman, D S
      Coffin, R S

        MeSH Terms

        • Animals
        • Cell Line
        • Cricetinae
        • Gene Deletion
        • Genes, Immediate-Early
        • Genetic Complementation Test
        • Genetic Vectors / genetics
        • Herpes Simplex Virus Protein Vmw65 / genetics
        • Herpes Simplex Virus Protein Vmw65 / physiology
        • Herpesvirus 1, Equid / genetics
        • Herpesvirus 1, Human / genetics
        • Herpesvirus 1, Human / growth & development
        • Horses
        • Humans
        • Immediate-Early Proteins / genetics
        • Promoter Regions, Genetic
        • Trans-Activators / genetics
        • Trans-Activators / physiology
        • Transcriptional Activation
        • Ubiquitin-Protein Ligases
        • Viral Proteins / genetics
        • Viral Proteins / physiology
        • Virion / physiology
        • Virus Assembly

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        Citations

        This article has been cited 8 times.
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