IRES-dependent replication of Venezuelan equine encephalitis virus makes it highly attenuated and incapable of replicating in mosquito cells

Eugenia Volkova, Elena Frolova, Justin R. Darwin, Naomi L. Forrester, Scott Weaver, Ilya Frolov

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The development of infectious cDNA for different alphaviruses opened an opportunity to explore their attenuation by extensively modifying the viral genomes, an approach that might minimize or exclude the reversion to the wild-type, pathogenic phenotype. Moreover, the genomes of such alphaviruses can be engineered to contain RNA elements that would be functional only in cells of vertebrate, but not insect, origin. In the present study, we developed a recombinant VEEV that is more attenuated than TC-83 and capable of replicating only in vertebrate cells. This phenotype was achieved by rendering the translation of the viral structural proteins, and ultimately viral replication, dependent on the internal ribosome entry site of encephalomyocarditis virus (EMCV IRES). This recombinant virus was viable, but required additional, adaptive mutations in nsP2 that strongly increased its replication rates. In spite of efficient replication in cultured vertebrate cells, the genetically modified VEEV demonstrated a highly attenuated phenotype in newborn mice, and yet induced protective immunity against VEEV infection.

Original languageEnglish (US)
Pages (from-to)160-169
Number of pages10
JournalVirology
Volume377
Issue number1
DOIs
StatePublished - Jul 20 2008

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Venezuelan Equine Encephalitis Viruses
Culicidae
Encephalomyocarditis virus
Alphavirus
Vertebrates
Phenotype
Viral Structural Proteins
Viral Genome
Insects
Cultured Cells
Immunity
Complementary DNA
Genome
RNA
Viruses
Mutation
Infection

Keywords

  • Alphavirus
  • EMCV IRES
  • Mosquito cells
  • Replication
  • VEEV

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

IRES-dependent replication of Venezuelan equine encephalitis virus makes it highly attenuated and incapable of replicating in mosquito cells. / Volkova, Eugenia; Frolova, Elena; Darwin, Justin R.; Forrester, Naomi L.; Weaver, Scott; Frolov, Ilya.

In: Virology, Vol. 377, No. 1, 20.07.2008, p. 160-169.

Research output: Contribution to journalArticle

Volkova, Eugenia ; Frolova, Elena ; Darwin, Justin R. ; Forrester, Naomi L. ; Weaver, Scott ; Frolov, Ilya. / IRES-dependent replication of Venezuelan equine encephalitis virus makes it highly attenuated and incapable of replicating in mosquito cells. In: Virology. 2008 ; Vol. 377, No. 1. pp. 160-169.
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