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NDT Advance Access published online on May 4, 2005
Nephrology Dialysis Transplantation, doi:10.1093/ndt/gfh783
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© The Author [2005]. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please email: journals.permissions@oupjournals.org
Received April 12, 2004
Accepted January 14, 2005

Original Articles

Gene transfer to the rat kidney in vivo and ex vivo using an adenovirus vector: factors influencing transgene expression

Jun Fujishiro 1, Shin-ichi Takeda 2, Yuichi Takeno 2, Koichi Takeuchi 3, Yukiyo Ogata 2, Masafumi Takahashi 2, Yoji Hakamata 2, Takashi Kaneko 2, Takashi Murakami 2, Takashi Okada 4, Keiya Ozawa 4, Kohei Hashizume 5, and Eiji Kobayashi 2*

1 Division of Organ Replacement Research, Center for Molecular Medicine, Tochigi, Japan; Department of Pediatric Surgery, Faculty of Medicine, University of Tokyo, Tokyo, Japan
2 Division of Organ Replacement Research, Center for Molecular Medicine, Tochigi, Japan
3 Division of Anatomy, Department of Anatomy, Jichi Medical School, Tochigi, Japan
4 Division of Genetic Therapeutics, Center for Molecular Medicine, Tochigi, Japan
5 Department of Pediatric Surgery, Faculty of Medicine, University of Tokyo, Tokyo, Japan

* To whom correspondence should be addressed.
Eiji Kobayashi, E-mail: eijikoba{at}jichi.ac.jp



  Abstract

Background. The characteristics of adenovirus-mediated gene transfer into the kidney are not well examined. We studied the effects of contact time and temperature on adenovirus-mediated transgene expression in rat kidneys, using catheter-based in vivo gene transfer and a rat renal transplant model ex vivo.

Methods. An adenovirus vector containing the luciferase (Ad-Luc) or {beta}-galactosidase (Ad-LacZ) gene was introduced in vivo into the kidney via a renal artery catheter. Various contact times and temperatures were evaluated. Ex vivo, the renal graft was injected with Ad-Luc through the renal artery, chilled for 60 min and then transplanted. Luciferase expression was evaluated periodically by a non-invasive bioimaging system or histology. Cells expressing the LacZ gene were identified by immunoelectron microscopy.

Results. In in vivo gene transfer, successful transgene expression was achieved; however, its efficiency was independent of contact time or temperature. In ex vivo gene transfer, transgene expression in the renal graft peaked early and gradually decreased. Strong gene expression was observed in the recipients' livers. LacZ expression was detected in fibroblasts, parietal epithelial cells of Bowman's capsule, mesangial cells, podocytes and tubular cells.

Conclusions. This study generated new information about in vivo and ex vivo gene transfer into the kidney, which would be useful for renal gene therapy.

Keywords: adenovirus; gene expression; gene transfer; renal transplantation.
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