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Mammalian orthoreovirus T3D infects U-118 MG cell spheroids independent of junction adhesion molecule-A

Gene Therapy volume 21, pages 609–617 (2014)Cite this article

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Abstract

In the canonical pathway, infection of cells by the wild-type mammalian orthoreovirus Type 3 Dearing (T3D) is dependent on the interaction of the viral spike protein σ1 with the high-affinity cellular receptor junction adhesion molecule-A (JAM-A). We previously demonstrated that the human glioblastoma cell line U-118 MG does not express JAM-A and resists reovirus T3D infection in standard cell culture conditions (SCCC). Heterologous JAM-A expression sensitises U-118 MG cells to reovirus T3D. Here we studied reovirus infection in U-118 MG cells grown in spheroid cultures with the premise that cells in such cultures resemble cells in tumours more than those grown under standard adherent cell culture conditions on a plastic surface. Although the U-118 MG cells in spheroids do not express JAM-A, they are susceptible to reovirus T3D infection. We show that this can be attributed to factors secreted by cells in the spheroids. The concentration of active extracellular proteases cathepsin B and L in the medium of spheroid cultures was increased 19- and 24-fold, respectively, as compared with SCCC. These enzymes can convert the reovirus particles into a form that can infect the U-118 MG cells independent of JAM-A. Taken together, these data demonstrate that infection of tumour cells by wild-type reovirus T3D is not strictly dependent on the expression of JAM-A on the cell surface.

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Acknowledgements

We thank Dr Martine Lamfers and Anne Kleijn (Department of Neurosurgery, Erasmus MC, Rotterdam, The Netherlands) for help in generating spheroids on solid agarose matrices. This work was supported in part by the European Union through the 6th Framework Program GIANT (Contract No. 512087).

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Authors and Affiliations

  1. Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands

    I J C Dautzenberg, D J M van den Wollenberg, S K van den Hengel, R W A Limpens, M Bárcena, A J Koster & R C Hoeben

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  1. I J C Dautzenberg
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  2. D J M van den Wollenberg
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Correspondence to R C Hoeben.

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Dautzenberg, I., van den Wollenberg, D., van den Hengel, S. et al. Mammalian orthoreovirus T3D infects U-118 MG cell spheroids independent of junction adhesion molecule-A. Gene Ther 21, 609–617 (2014). https://doi.org/10.1038/gt.2014.34

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