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Picolinic acid- or desferrioxamine-inducible autocrine activation of macrophages engineered to produce IFNγ: an approach for gene therapy

Gene Therapy volume 11, pages 560–568 (2004)Cite this article

Abstract

Macrophage (Mφ)-based vectors are highly mobile cellular shuttles designed to deliver therapeutic genes within the tissues. We engineered a mouse Mφ cell line to express the murine interferon-γ (IFNγ) under the control of an inducible promoter containing the hypoxia-responsive element, which can be triggered by hypoxia and other stimuli. We show that this Mφ vector can be induced to produce IFNγ under normoxic conditions by stimulation with picolinic acid (PA), a catabolite of tryptophan, or desferrioxamine (DFX), an iron-chelating drug. The Mφ vector responds to IFNγ with the induction of IRF-1 and of other IFNγ-inducible genes, the expression of Ia antigens and induction of phagocytic activity. Inducible nitric oxygen synthase gene expression, nitric oxide production, as well as TNFα secretion were enhanced by PA or DFX as the sole stimuli. None of the above responses could be triggered individually by PA or DFX in control, normal Mφ, indicating that the Mφ vector overcame the need for costimulatory molecules derived from the immune system for its full activation. Furthermore, we demonstrate that extracellular iron can downregulate such response, thereby identifying an additional tool for the fine tuning of the Mφ vector response to stimulation.

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Acknowledgements

We thank Ms Chantal Dabizzi for secretarial assistance. This work was supported by grants from the Italian Association for Cancer Research (AIRC), Fondazione Italiana per la Lotta al Neuroblastoma, Associazione Italiana Glicogenosi and San Paolo foundation. Dr Pastorino was supported by a fellowship from the Fondazione Italiana per la Ricerca sul Cancro (FIRC).

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Author notes

  1. S Pastorino

    Present address: Neuro Oncology Branch, National Cancer Institute, National Institute of Health, 36 Convent Drive, Bldg 36, Rm 3B02, Bethesda, MD, 20892, USA

  2. L Carta

    Present address: Brookdale Department of Molecular, Cell and Developmental Biology, Mount Sinai School of Medicine, 1 G. Levy Place, NY, 10029, New York, USA

  3. S Pastorino and L Carta: The first two authors, contributed equally to this work

Authors and Affiliations

  1. Laboratory of Molecular Biology, G Gaslini Institute, Largo Gaslini, Genova, Italy

    S Pastorino, L Carta, M Puppo, MC Bosco & L Varesio

  2. DTP-Tumor Hypoxia Program, National Cancer Institute, National Institute of Health, Frederick, MD, USA

    G Melillo

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Pastorino, S., Carta, L., Puppo, M. et al. Picolinic acid- or desferrioxamine-inducible autocrine activation of macrophages engineered to produce IFNγ: an approach for gene therapy. Gene Ther 11, 560–568 (2004). https://doi.org/10.1038/sj.gt.3302217

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