title: Gene transfer to the rat kidney in vivo and ex vivo using an adenovirus vector: factors influencing transgene expression.
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.
authors.affiliation: Division of Organ Replacement Research, Center for Molecular Medicine, Jichi Medical School, 3311-1 Yakushiji, Minamikawachi-machi, Tochigi 329-0498, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Fujishiro
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Takeda
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Takeno
authors.forename: Koichi
authors.initials: K
authors.lastname: Takeuchi
authors.forename: Yukiyo
authors.initials: Y
authors.lastname: Ogata
authors.forename: Masafumi
authors.initials: M
authors.lastname: Takahashi
authors.forename: Yoji
authors.initials: Y
authors.lastname: Hakamata
authors.forename: Takashi
authors.initials: T
authors.lastname: Kaneko
authors.forename: Takashi
authors.initials: T
authors.lastname: Murakami
authors.forename: Takashi
authors.initials: T
authors.lastname: Okada
authors.forename: Keiya
authors.initials: K
authors.lastname: Ozawa
authors.forename: Kohei
authors.initials: K
authors.lastname: Hashizume
authors.forename: Eiji
authors.initials: E
authors.lastname: Kobayashi
mesh_terms: D000256:Adenoviridae; D000818:Animals; D015870:Gene Expression / Q000502:physiology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007668:Kidney / Q000502:physiology*; D007763:Lac Operon / Q000502:physiology*; D049409:Luciferases, Firefly / Q000378:metabolism*; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D013696:Temperature; D013997:Time Factors; D019076:Transgenes / Q000502:physiology
chemical_list: D049409:Luciferases, Firefly