Expression of vaccinia E3L and K3L genes by a novel recombinant canarypox HIV vaccine vector enhances HIV-1 pseudovirion production and inhibits apoptosis in human cells.
Poxviruses that are attenuated for growth in human cells provide a safe means of HIV antigen expression and are capable of eliciting HIV-specific immune responses, including CD8+ cytotoxic T-lymphocyte (CTL) responses. HIV-1 antigen expression in human cells by attenuated poxvirus vectors may be limited by interferon-mediated host defense mechanisms. To enhance HIV antigen expression in human cells, the vaccinia virus E3L and K3L genes were inserted into a canarypox vector that expresses HIV-1 Gag, Env, and a Nef/Pol polyepitope string. E3L and K3L markedly reduced the activation of the double-stranded RNA-dependent protein kinase, PKR, and led to a significant reduction in apoptosis in HeLa cells. Production and release of HIV-1 antigen in the form of pseudovirions was enhanced in both duration and magnitude by this vector modification. The addition of immunomodulatory genes to attenuated poxviruses represents a novel strategy for enhancing antigen production by live vector HIV vaccine candidates.
Fang
Z Y
ZY
Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, TN 37232-2581, USA.
Limbach
K
K
Tartaglia
J
J
Hammonds
J
J
Chen
X
X
Spearman
P
P
D017209:Apoptosis; D028121:Canarypox virus / Q000235:genetics*; D015870:Gene Expression; D005814:Genes, Viral; D015752:Genes, env; D015750:Genes, gag; D015754:Genes, nef; D015751:Genes, pol; D005822:Genetic Vectors / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D010766:Phosphorylation; D016601:RNA-Binding Proteins / Q000235:genetics*; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*; D014771:Virion / Q000502:physiology; D019065:Virus Assembly; D019892:eIF-2 Kinase / Q000378:metabolism
C081374:E3L protein, Vaccinia virus; D016601:RNA-Binding Proteins; D014764:Viral Proteins; C081646:pK3 protein, Vaccinia virus; D019892:eIF-2 Kinase