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A new genetic method to generate and isolate small, short-lived but highly potent dendritic cell-tumor cell hybrid vaccines

Nature Medicine volume 9, pages 1215–1219 (2003)Cite this article

Abstract

Fusion of tumor cells with antigen-presenting cells (APCs) has been proposed for the preparation of cancer vaccines. However, generation of these hybrids, using physical or chemical methods such as electrofusion or polyethylene glycol (PEG), has been difficult to standardize. Characterization of cell fusion has also been problematic because of difficulties in differentiating fusion from cell aggregation, leakage of cellular dyes and dendritic-cell (DC) phagocytosis of tumor material. In this report, we describe a new method to generate hybrid cell vaccines, based on gene transfer of a viral fusogenic membrane glycoprotein (FMG) into tumor cells, and incorporate a genetic method by which true hybrid formation can be unambiguously detected. We describe a new class of tumor cell–DC hybrid that can be rapidly isolated after cell fusion. These hybrids are highly potent in in vitro antigen presentation assays, target lymph nodes in vivo and are powerful immunogens against established metastatic disease.

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Figure 1: A genetic method to identify and isolate DC–tumor cell hybrids.
Figure 2: Hybrid cells can be isolated by FACS and express both tumor and DC markers early, but not late, after fusion.
Figure 3: Small, GFP-positive hybrid cells retain the ability to travel to lymph nodes.
Figure 4: Small FMG-induced tumor cell–DC hybrids are very efficient at presenting a model antigen to T cells.
Figure 5: Small, short-lived GFP-positive hybrids are effective immunogens against 7-d established metastatic disease.

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Acknowledgements

We thank T. Higgins for expert secretarial assistance. This work was supported by National Institutes of Health grants RO1 CA85931, RO1 CA094180 and P50 CA91956; AstraZeneca; the Mayo Foundation; and the Université Libre de Bruxelles.

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Authors and Affiliations

  1. Molecular Medicine Program, Mayo Clinic, 200 First Street SW, Rochester, 55905, Minnesota, USA

    Vy Phan, Tim Kottke, Michael Gough, Sharon Altmann, Andrew Bateman & Richard Vile

  2. CRUK Oncology Unit, St. James' Hospital, Leeds, LS9 7TF, UK

    Fiona Errington & Alan Melcher

  3. Université Libre de Bruxelles, Gosselies, 6041, Belgium

    S Chiat Cheong & Annick Brandenburger

  4. AstraZeneca, Alderley Park, Cheshire, SK10, UK

    Steve Emery

  5. Department of Otorhinolaryngology, Mayo Clinic, 200 First Street SW, Rochester, 55905, Minnesota, USA

    Scott Strome

  6. INSERM U517, Faculty of Medicine, Dijon, 21079, Cedex, France

    Bernard Bonnotte

  7. Department of Immunology, Mayo Clinic, 200 First Street SW, Rochester, 55905, Minnesota, USA

    Richard Vile

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  1. Vy Phan
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Correspondence to Richard Vile.

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Phan, V., Errington, F., Cheong, S. et al. A new genetic method to generate and isolate small, short-lived but highly potent dendritic cell-tumor cell hybrid vaccines. Nat Med 9, 1215–1219 (2003). https://doi.org/10.1038/nm923

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