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  • Viral Transfer Technology
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Calcium phosphate precipitates augment adenovirus-mediated gene transfer to blood vessels in vitro and in vivo

Gene Therapy volume 7, pages 1284–1291 (2000)Cite this article

Abstract

Adenovirus (Ad)-mediated gene transfer to blood vessels is relatively inefficient, probably because binding of adenovirus to the endothelium and adventitia seems to be limited. Association of calcium phosphate (CaPi) precipitates with adenovirus improves efficiency of gene transfer to some cells in culture and to mouse lung in vivo. In this study, we tested the hypothesis that CaPi is useful for adenovirus-mediated gene transfer to blood vessels. In fibroblast and endothelial cells in culture, Ad:CaPi coprecipitates greatly increased transgene expression. Ad:CaPi also enhanced transgene expression in both adventitia and endothelium of carotid arteries and aortae from rabbits studied ex vivo. After injection of Ad:CaPi into the cisterna magna of rabbits in vivo, the transgene product was markedly increased in leptomeninges of the ventral brain stem, including the adventitia of the basilar artery. We also examined mechanisms of enhanced gene transfer. Binding of adenovirus to fibroblast and endothelial cells in culture, and to the basilar artery in vivo, as determined using Southern blot analysis, was augmented by CaPi. Antibody to adenoviral fiber knob did not inhibit augmented transgene expression by Ad:CaPi. The finding suggests that improved adenoviral binding occurs primarily via a fiber-independent pathway. Thus, CaPi precipitates are useful for improvement of adenovirus-mediated gene transfer to blood vessels in vitro and in vivo.

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Acknowledgements

We thank Dr Yi Chu and Robert W Walters for excellent advice, Pamela K Tompkins and Sonya Mehta for technical assistance, and Arlinda LaRose for typing the manuscript. We also thank Dr Beverly L Davidson, Richard D Anderson, and the University of Iowa Gene Transfer Vector Core for preparation of virus. This work was supported by HL 14388, HL 16066, NS 24621, HL 14355, NS 37386, HK 62984, and DK 54759.

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Authors and Affiliations

  1. Department of Internal Medicine, Pharmacology, University of Iowa College of Medicine, and Veterans Administration Medical Center, Iowa City, Iowa, USA

    K Toyoda, JJ Andresen, FM Faraci & DD Heistad

  2. Cardiovascular Center, University of Iowa College of Medicine, and Veterans Administration Medical Center, Iowa City, Iowa, USA

    K Toyoda, JJ Andresen, FM Faraci & DD Heistad

  3. Department of Physiology and Biophysics, University of Iowa College of Medicine, and Veterans Administration Medical Center, Iowa City, Iowa, USA

    J Zabner

  4. Howard Hughes Medical Institute, University of Iowa College of Medicine, and Veterans Administration Medical Center, Iowa City, Iowa, USA

    J Zabner

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Toyoda, K., Andresen, J., Zabner, J. et al. Calcium phosphate precipitates augment adenovirus-mediated gene transfer to blood vessels in vitro and in vivo. Gene Ther 7, 1284–1291 (2000). https://doi.org/10.1038/sj.gt.3301214

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