Generation of high-titer retroviral vector-producing macrophages as vehicles for in vivo gene transfer

S. Pastorino, S. Massazza, M. Cilli, L. Varesio, M. C. Bosco

Research output: Contribution to journalArticlepeer-review


The goal of this project was to develop a novel gene transfer system based on macrophages (Mφ) as shuttles of recombinant retroviral vectors carrying therapeutic or marker genes. The murine Mφ cell line WGL5 was used as a source of Mφ for this study. We generated retrovirus-producing Mφ by transducing the WGL5 cells with a replication-defective retroviral vector carrying the enhanced green fluorescent protein (EGFP) reporter gene and the Moloney murine leukemia virus (MoMLV) as helper virus. We demonstrated stable integration of the recombinant retrovirus in the Mφ genome, efficient recombinant retrovirus production, and EGFP gene delivery to different cell lines in vitro. To evaluate Mφ-mediated EGFP gene transfer in vivo, allogeneic mice were injected s.c. with the retrovirus-producing WGL5 Mφ, that gave rise to solid tumor masses at the injection site, highly infiltrated with host leukocytes. We observed EGFP fluorescence in tumor-infiltrating CD4+ and Cd8+ host T lymphocytes, providing direct evidence of the ability of engineered Mφ to mediate EGFP gene delivery to host cells in vivo. Moreover, we showed that retrovirus-producing Mφ could home to different organs in vivo following i.v. injection into mice. These data demonstrate that Mφ can be engineered as cellular vehicles for recombinant retroviruses carrying heterologous genes and suggest potential applications of this novel vector system for gene therapy.

Original languageEnglish
Pages (from-to)431-441
Number of pages11
JournalGene Therapy
Issue number6
Publication statusPublished - 2001


  • Gene therapy
  • In vivo transduction
  • Macrophages
  • Retrovirus

ASJC Scopus subject areas

  • Genetics


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