title: 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.
abstract: 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.
authors.affiliation: Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, TN 37232-2581, USA.
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Fang
authors.forename: K
authors.initials: K
authors.lastname: Limbach
authors.forename: J
authors.initials: J
authors.lastname: Tartaglia
authors.forename: J
authors.initials: J
authors.lastname: Hammonds
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.forename: P
authors.initials: P
authors.lastname: Spearman
mesh_terms: 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
chemical_list: C081374:E3L protein, Vaccinia virus; D016601:RNA-Binding Proteins; D014764:Viral Proteins; C081646:pK3 protein, Vaccinia virus; D019892:eIF-2 Kinase