Analysis of Venezuelan equine encephalitis replicon particles packaged in different coats

The Venezuelan equine encephalitis (VEE) virus replicon system was used to produce virus-like replicon particles (VRP) packaged with a number of different VEE-derived glycoprotein (GP) coats. The GP coat is believed to be responsible for the cellular tropism noted for VRP and it is possible that dif...

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Bibliographic Details
Published in:PloS one 2008-07, Vol.3 (7), p.e2709-e2709
Main Authors: Kamrud, Kurt I, Alterson, Kim D, Andrews, Chasity, Copp, Laura O, Lewis, Whitney C, Hubby, Bolyn, Patel, Deepa, Rayner, Jonathan O, Talarico, Todd, Smith, Jonathan F
Format: Article
Language:English
Subjects:
Analysis
Animals
Biotechnology
Cell culture
Cloning
Coat protein
Coating
Cytomegalovirus
Dendritic cells
Encephalitis
Encephalitis Virus, Venezuelan Equine - metabolism
Encephalomyelitis, Venezuelan Equine - virology
Enzyme-Linked Immunosorbent Assay
Equine encephalomyelitis
Female
Genes
Genetic Vectors
Genome
Genomes
Genomics
Glycoproteins
Glycoproteins - chemistry
Health aspects
Heparan sulfate
HIV
Human immunodeficiency virus
Immune response (cell-mediated)
Immune response (humoral)
Immune System
Immunization
In vivo methods and tests
Infectious Diseases/Viral Infections
Infectivity
Kinases
Mice
Mice, Inbred BALB C
Mutation
Phosphorylation
Proteins
Replicon
Reverse Transcriptase Polymerase Chain Reaction
Tropism
Venezuelan equine encephalitis
Virology
Viruses
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Description
Summary:The Venezuelan equine encephalitis (VEE) virus replicon system was used to produce virus-like replicon particles (VRP) packaged with a number of different VEE-derived glycoprotein (GP) coats. The GP coat is believed to be responsible for the cellular tropism noted for VRP and it is possible that different VEE GP coats may have different affinities for cells. We examined VRP packaged in four different VEE GP coats for their ability to infect cells in vitro and to induce both humoral and cellular immune responses in vivo. The VRP preparations were characterized to determine both infectious units (IU) and genome equivalents (GE) prior to in vivo analysis. VRP packaged with different VEE GP coats demonstrated widely varying GE/IU ratios based on Vero cell infectivity. BALB/c mice were immunized with the different VRP based on equal GE titers and the humoral and cellular responses to the expressed HIV gag gene measured. The magnitude of the immune responses measured in mice revealed small but significant differences between different GP coats when immunization was based on GE titers. We suggest that care should be taken when alternative coat proteins are used to package vector-based systems as the titers determined by cell culture infection may not represent accurate particle numbers and in turn may not accurately represent actual in vivo dose.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0002709