Structure and immunogenicity of alternative forms of the simian immunodeficiency virus gag protein expressed using Venezuelan equine encephalitis virus replicon particles

Abstract Venezuelan equine encephalitis virus replicon particles (VRP) were engineered to express different forms of SIV Gag to compare expression in vitro , formation of intra- and extracellular structures and induction of humoral and cellular immunity in mice. The three forms examined were full-le...

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Bibliographic Details
Published in:Virology (New York, N.Y.) N.Y.), 2007-06, Vol.362 (2), p.362-373
Main Authors: Cecil, Chad, West, Ande, Collier, Martha, Jurgens, Christy, Madden, Victoria, Whitmore, Alan, Johnston, Robert, Moore, Dominic T, Swanstrom, Ronald, Davis, Nancy L
Format: Article
Language:English
Subjects:
Animals
Antibodies, Viral - blood
Cell Line
Cells, Cultured
Cercopithecus aethiops
Embryo, Mammalian - cytology
Encephalitis Virus, Venezuelan Equine - genetics
Enzyme-Linked Immunosorbent Assay
Female
Fibroblasts
gene expression
Gene Products, gag - chemistry
Gene Products, gag - immunology
genetic vectors
Genetic Vectors - genetics
H-2 Antigens - immunology
Infectious Disease
Interferon-gamma - biosynthesis
Lymphocytes - immunology
Mice
Mice, Inbred BALB C
Mice, Inbred Strains
Microscopy, Electron, Transmission
Models, Animal
nucleocapsid proteins
Pregnancy
replicon
Replicon - genetics
Simian immunodeficiency virus
Simian Immunodeficiency Virus - immunology
SIV Gag
SIV Gag-specific immunity in BALB/c mice
Vaccines, Synthetic - genetics
Vaccines, Synthetic - immunology
VEE replicon vectors
Venezuelan equine encephalitis virus
Vero Cells
Viral Vaccines - genetics
Viral Vaccines - immunology
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Summary:Abstract Venezuelan equine encephalitis virus replicon particles (VRP) were engineered to express different forms of SIV Gag to compare expression in vitro , formation of intra- and extracellular structures and induction of humoral and cellular immunity in mice. The three forms examined were full-length myristylated SIV Gag (Gagmyr+ ), full-length Gag lacking the myristylation signal (Gagmyr− ) or a truncated form of Gagmyr− comprising only the matrix and capsid domains (MA/CA). Comparison of VRP-infected primary mouse embryo fibroblasts, mouse L929 cells and primate Vero cells showed comparable expression levels for each protein, as well as extracellular virus-like particles (VRP-Gagmyr+ ) and distinctive cytoplasmic aggregates (VRP-Gagmyr− ) with each cell type. VRP were used to immunize BALB/c mice, and immune responses were compared using an interferon (IFN)-γ ELISPOT assay and a serum antibody ELISA. Although all three VRP generated similar levels of IFN-γ-producing cells at 1 week post-boost, at 10 weeks post-boost the MA/CA-VRP-induced response was maintained at a significantly higher level relative to that induced by Gagmyr+ -VRP. Antibody responses to MA/CA-VRP and Gagmyr+ -VRP were not significantly different.
ISSN:0042-6822
1096-0341
DOI:10.1016/j.virol.2006.12.029