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Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase

Gene Therapy volume 12, pages 169–176 (2005)Cite this article

Abstract

The treatment of solid tumors by retroviral delivery of the herpes simplex virus thymidine kinase (TK) followed by ganciclovir (GCV) treatment has so far shown only limited success in patients. One major drawback in this approach is the lack of efficient in vivo gene delivery to cancer cells. Although, the transduction of every single tumor cell is not a requirement since the bystander effect (BE) mediated by gap junctions allows the diffusion of the toxic GCV metabolites from TK-expressing cells toward untransduced cells. To render the TK/GCV approach more potent, and independent of the level of gap junctions, we have tested the efficiency of a TK mutant (TK30) fused to VP22, a herpes simplex protein that seems to be capable of intercellular trafficking. We failed to detect an increase in the BE with cells expressing VP22 fused to TK30 versus cells containing TK30 alone, and this result forced us to reinvestigate the trafficking properties of VP22. Using very sensitive Western blot and fluorescence assays, we were not able to detect the spread of VP22 fused either to TK30 or GFP. These results indicate that VP22 cannot be used as a cargo to translocate TK30 or GFP.

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Acknowledgements

This study was supported by grants from the Canadian Institute of Health Research (IC074582) and Manuel Caruso is a Junior II Research Scholar of the Fonds de la Recherche en Santé du Québec. Vincent Roy holds a graduate student award from the MCETC/CIHR strategic training program. We are grateful to Margaret Black for providing us with TK30 and the rabbit polyclonal antibody directed against TK. We thank Nancy Roberge for the help in flow cytometry and the cell sorting.

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  1. J Qiao

    Present address: Molecular Medicine Program, Mayo Clinic, Rochester, MN, 55905, USA

  2. V Roy and J Qiao: These authors contributed equally to this work

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  1. Le Centre de Recherche en Cancérologie de l'Université Laval, L'Hôtel-Dieu de Québec, Centre Hospitalier Universitaire de Québec, Québec, Canada

    V Roy, J Qiao, P de Campos-Lima & M Caruso

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Roy, V., Qiao, J., de Campos-Lima, P. et al. Direct evidence for the absence of intercellular trafficking of VP22 fused to GFP or to the herpes simplex virus thymidine kinase. Gene Ther 12, 169–176 (2005). https://doi.org/10.1038/sj.gt.3302394

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