Antigen-mediated growth control of hybridoma cells via a human artificial chromosome

Biochim Biophys Acta. 2007 Feb;1770(2):206-12. doi: 10.1016/j.bbagen.2006.10.014. Epub 2006 Nov 6.

Abstract

Human artificial chromosome (HAC) vectors possess several characteristics sufficient for the requirements of gene therapy vectors, including stable episomal maintenance and mediation of long-term transgene expression. In this study, we adopted an antigen-mediated genetically modified cell amplification (AMEGA) system employing an antibody/cytokine receptor chimera that triggers a growth signal in response to a cognate non-toxic antigen, and applied it to growth control of HAC-transferred cells by adding an antigen that differed from cytokines that may manifest pleiotropic effects. We previously constructed a novel HAC vector, 21 Delta qHAC, derived from human chromosome 21, housed in CHO cells. Here, we constructed an HAC vector harboring an ScFv-gp130 chimera responsive to fluorescein-conjugated BSA (BSA-FL) as well as a model transgene, enhanced green fluorescent protein (EGFP), in CHO cells. The modified HAC was transferred into interleukin (IL)-6-dependent hybridoma 7TD1 cells by microcell-mediated chromosome transfer, and the cells were subsequently found to show BSA-FL-dependent cell growth and sustained expression of EGFP in the absence of IL-6. The AMEGA system in combination with HAC technology will be useful for increasing the efficacy of gene therapy by conferring a growth advantage on the genetically modified cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cell Division / physiology*
  • Cell Line
  • Chromosomes, Artificial, Human / physiology*
  • Cricetinae
  • Cricetulus
  • Gene Transfer Techniques
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Humans
  • Hybridomas / cytology*
  • Interleukin-6 / pharmacology
  • Mice
  • Recombinant Fusion Proteins
  • Transfection

Substances

  • Interleukin-6
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins