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The αvβ5 integrin of hematopoietic and nonhematopoietic cells is a transduction receptor of RGD-4C fiber-modified adenoviruses

Gene Therapy volume 10, pages 1643–1653 (2003)Cite this article

Abstract

Epithelial and endothelial cells expressing the primary Coxsackie virus B adenovirus (Ad) receptor (CAR) and integrin coreceptors are natural targets of human Ad infections. The fiber knob of species A, C, D, E and F Ad serotypes binds CAR by mimicking the CAR–homodimer interface, and the penton base containing arginine–glycine–aspartate (RGD) motifs binds with low affinity to αv integrins inducing cell activation. Here, we generated seven different genetically modified Ad vectors with RGD sequences inserted into the HI loop of fiber knob. All mutants bound and infected CAR and αv integrin-positive epithelial cells with equal efficiencies. However, the Ads containing two additional cysteines, both N and C terminals of the RGD sequence (RGD-4C), were uniquely capable of transducing CAR-less hematopoietic and nonhematopoietic human tumor cell lines and primary melanoma cells. Both binding and transduction of RGD-4C Ad were blocked by soluble RGD peptides. Flow cytometry of cell surface integrins and virus binding to CAR-less cells in the presence of function-blocking anti-integrin antibodies indicated that the αvβ5 integrin, but not αvβ3, αIIbβ3 or β1,α5 or α6-containing integrins served as a functional transduction receptor of the RGD-4C Ads. However, in cells with low levels of αvβ5 integrin, the function-blocking anti-αvβ5 antibodies were not effective, unlike soluble RGD peptides. Collectively, our data demonstrate that the αvβ5 integrin is a functional transduction receptor of RGD-4C Ads in the absence of CAR, and that additional RGD receptors are targets of these viruses. The RGD-4C vectors further extend the tropism of Ads towards potential human therapies.

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Acknowledgements

This work has been supported by the Kanton Zürich and by a grant of the Krebsliga of the Kanton Zürich (to SH). UFG was supported by the Swiss National Science Foundation. We thank C Torres-de los Reyes and E Horvath for excellent technical assistance and F Ochsenbein for graphic designs.

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  1. H Nagel and S Maag: The first two authors contributed equally

Authors and Affiliations

  1. Institute of Molecular Biology, University of Zürich, Zürich, Switzerland

    H Nagel, S Maag & S Hemmi

  2. Department of Dermatology, University of Zürich Medical School, Zürich, Switzerland

    A Tassis & F O Nestlé

  3. Institute of Zoology, University of Zürich, Zürich, Switzerland

    U F Greber

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  1. H Nagel
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Nagel, H., Maag, S., Tassis, A. et al. The αvβ5 integrin of hematopoietic and nonhematopoietic cells is a transduction receptor of RGD-4C fiber-modified adenoviruses. Gene Ther 10, 1643–1653 (2003). https://doi.org/10.1038/sj.gt.3302058

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