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

Virology. 2001 Dec 20;291(2):272-84. doi: 10.1006/viro.2001.1209.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Apoptosis*
  • Canarypox virus / genetics*
  • Gene Expression*
  • Genes, Viral
  • Genes, env
  • Genes, gag
  • Genes, nef
  • Genes, pol
  • Genetic Vectors / genetics*
  • HIV-1 / genetics
  • HIV-1 / physiology*
  • HeLa Cells
  • Humans
  • Microscopy, Electron
  • Phosphorylation
  • RNA-Binding Proteins / genetics*
  • Recombination, Genetic
  • Vaccinia virus / genetics*
  • Viral Proteins / genetics*
  • Virion / physiology
  • Virus Assembly*
  • eIF-2 Kinase / metabolism

Substances

  • E3L protein, Vaccinia virus
  • RNA-Binding Proteins
  • Viral Proteins
  • pK3 protein, Vaccinia virus
  • eIF-2 Kinase