Sequestration of adenoviral vector by Kupffer cells leads to a nonlinear dose response of transduction in liver

Mol Ther. 2001 Jan;3(1):28-35. doi: 10.1006/mthe.2000.0227.


Systemic administration of a recombinant adenovirus encoding the human interferon-beta gene (H5.110CMVhIFN-beta) results in transduction of hepatocytes and detectable circulating levels of IFN-beta protein. In preclinical studies in mice, we noticed a distinctly nonlinear dose response, with low levels of virus (1-3 x 10(10) viral particles) yielding barely detectable levels of IFN-beta but with a higher viral dose (1 x 10(11) particles) resulting in disproportionately high IFN-beta levels. Further studies showed that transgene expression levels from low viral doses could be dramatically enhanced by coadministering an unrelated recombinant adenovirus (H5.110CMVlacZ), suggesting that there was a viral dose threshold effect for efficient viral transduction and/or IFN-beta expression. This enhancement of reporter expression by a nonreporter adenovirus, effective upon coadministration, was further enhanced by preadministration of H5.110CMVlacZ (up to 8 h), but was ineffective if the helper virus was administered as little as 5 min after the H5.110CMVhIFN-beta reporter virus. Our data suggest that the reticuloendothelial system plays a role in this threshold effect, such that low doses of virus are efficiently taken up by the RES/Kupffer cells without leading to appreciable transgene expression, whereas high doses saturate these cells and are able to productively transduce hepatocytes. A better understanding of this phenomenon could have an impact on gene therapy clinical trial safety and efficacy.

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Carbocyanines / pharmacology
  • Dose-Response Relationship, Drug
  • Endothelium / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Fluorescent Dyes / pharmacology
  • Genes, Reporter
  • Genetic Vectors*
  • Hepatocytes / metabolism
  • Humans
  • Interferon-beta / genetics
  • Interferon-beta / metabolism
  • Kupffer Cells / metabolism*
  • Liver / metabolism*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Mice, Nude
  • Species Specificity
  • Tissue Distribution
  • Transduction, Genetic
  • Transgenes / genetics
  • Tumor Cells, Cultured
  • alpha 1-Antitrypsin / genetics
  • alpha 1-Antitrypsin / metabolism
  • beta-Galactosidase / metabolism


  • Carbocyanines
  • Fluorescent Dyes
  • alpha 1-Antitrypsin
  • cyanine dye 3
  • Interferon-beta
  • beta-Galactosidase