Vaccinia virus infection and gene transduction in cultured neurons

Gregory A. Allen, B. Dénes, Istvan Fodor, Marino De Leon

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The study of neurons in culture would benefit from the development of a gene transduction system capable of delivering foreign genes at high efficiency, as transduction of primary neurons with existing systems is inefficient. The efficacy of lytic vaccinia virus (VV) infection of primary retinal cultures and PC12 cells (a model of neuronal differentiation) was examined in order to determine the efficiency of gene transduction using VV in neuronal primary culture. VV was able to infect retinal cells and PC12 cells and express transgenes of Escherichia coli β-galactosidase (lacZ) and epithelial fatty acid binding protein (E-FABP) in a virus dose-dependent manner. Most (50-100%) of the retinal cells were positive for transgene protein at multiplicities of infection (MOI) between 10 and 100 plaque-forming units (PFU), while over 50% of VV-infected PC12 cells expressed the virus encoded gene at an MOI = 10. The production of foreign mRNA and protein by VV following infection was verified by PCR and Western blot. Because VV is a lytic virus, cytopathic effects were examined. Retinal cultures maintained for 0.5 days in vitro showed greater than 90% survival at 24 h post-infection, while 14-day cultures were equally viable for 48 h. Retinal ganglion cells and differentiated PC12 cells appear to be more protected against lytic VV infection than proliferating glial and undifferentiated PC12 cells. These data suggest that VV may be a useful vector for delivering foreign genes to neuronal cells with an efficient transient transgene expression.

Original languageEnglish
Pages (from-to)1087-1096
Number of pages10
JournalMicrobes and Infection
Volume7
Issue number9-10
DOIs
Publication statusPublished - Jul 2005

Fingerprint

Vaccinia virus
Virus Diseases
PC12 Cells
Neurons
Genes
Transgenes
Viruses
Infection
Galactosidases
Fatty Acid-Binding Proteins
Retinal Ganglion Cells
Neuroglia
Proteins
Western Blotting
Escherichia coli
Polymerase Chain Reaction
Messenger RNA

Keywords

  • Gene expression
  • PC12
  • Retinal ganglion cells
  • Vaccinia

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Vaccinia virus infection and gene transduction in cultured neurons. / Allen, Gregory A.; Dénes, B.; Fodor, Istvan; De Leon, Marino.

In: Microbes and Infection, Vol. 7, No. 9-10, 07.2005, p. 1087-1096.

Research output: Contribution to journalArticle

Allen, Gregory A. ; Dénes, B. ; Fodor, Istvan ; De Leon, Marino. / Vaccinia virus infection and gene transduction in cultured neurons. In: Microbes and Infection. 2005 ; Vol. 7, No. 9-10. pp. 1087-1096.
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