Construction of stable producer cells to make high-titer lentiviral vectors for dendritic cell-based vaccination

Biotechnol Bioeng. 2012 Jun;109(6):1551-60. doi: 10.1002/bit.24413. Epub 2012 Jan 17.


Lentiviral vectors (LVs) enveloped with an engineered Sindbis virus glycoprotein can specifically bind to dendritic cells (DCs) through the surface receptor DC-SIGN and induce antigen expression, thus providing an efficient method for delivering DC-directed vaccines. In this study, we constructed a stable producer line (LV-MGFP) for synthesizing DC-SIGN-targeted HIV-1-based LVs (DC-LVs) encoding green fluorescent protein (GFP) by a concatemeric array transfection technique. We demonstrated that the established stable clones could routinely produce vector supernatants with titers above 10(7) transduction units per milliliter (TU/mL) during a continuous 3-month cell passage. The producer cells were also capable of generating similar titers of DC-LVs in serum-free medium. Moreover, the addition of 1-deoxymannojirimycin (DMJ) enabled the producer cells to manufacture DC-LVs with both improved titers and enhanced potency to evoke antigen-specific CD8(+) T cell responses in mice. The stable lines could accommodate the replacement of the internal murine stem cell virus (MSCV) promoter with the human ubiquitin-C (Ubi) promoter in the lentiviral backbone. The resulting DC-LVs bearing Ubi exhibited the enhanced potency to elicit vaccine-specific immunity. Based on accumulated evidence, our studies support the application of this production method in manufacturing DC-LVs for preclinical and clinical testing of novel DC-based immunization.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotechnology / methods*
  • CD8-Positive T-Lymphocytes / immunology
  • Cell Line
  • Culture Media, Serum-Free
  • Dendritic Cells / immunology*
  • Genes, Reporter
  • Genetic Vectors*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HIV-1 / genetics*
  • Mice
  • Sindbis Virus / genetics
  • Staining and Labeling
  • Technology, Pharmaceutical / methods*
  • Tissue Culture Techniques / methods
  • Transduction, Genetic*
  • Vaccination / methods*
  • Viral Envelope Proteins / genetics
  • Viral Load


  • Culture Media, Serum-Free
  • Viral Envelope Proteins
  • Green Fluorescent Proteins