title: Evaluation of permissiveness and cytotoxic effects in equine chondrocytes, synovial cells, and stem cells in response to infection with adenovirus 5 vectors for gene delivery.
abstract: OBJECTIVE
  To evaluate host cell permissiveness and cytotoxic effects of recombinant and modified adenoviral vectors in equine chondrocytes, synovial cells, and bone marrow-derived mesenchymal stem cells (BMD-MSCs).
  
  
  METHODS
  Articular cartilage, synovium, and bone marrow from 15 adult horses.
  
  
  METHODS
  Equine chondrocytes, synovial cells, and BMD-MSCs and human carcinoma (HeLa) cells were cultured and infected with an E-1-deficient adenovirus vector encoding the beta-galactosidase gene or the green fluorescent protein gene (Ad-GFP) and with a modified E-1-deficient vector with the arg-gly-asp capsid peptide insertion and containing the GFP gene (Ad-RGD-GFP). Percentages of transduced cells, total and transduced cell counts, and cell viability were assessed 2 and 7 days after infection.
  
  
  RESULTS
  -Permissiveness to adenoviral vector infection was significantly different among cell types and was ranked in decreasing order as follows: HeLa cells > BMD-MSCs > chondrocytes > synovial cells. Morphologic signs of cytotoxicity were evident in HeLa cells but not in equine cells. Numbers of transduced cells decreased by day 7 in all cell types except equine BMD-MSCs. Transduction efficiency was not significantly different between the Ad-GFP and Ad-RGD-GFP vectors.
  
  
  CONCLUSIONS
  Sufficient gene transfer may be achieved by use of an adenovirus vector in equine cells. High vector doses can be used in equine cells because of relative resistance to cytotoxic effects in those cells. Greater permissiveness and sustained expression of transgenes in BMD-MSCs make them a preferential cell target for gene therapy in horses.
authors.affiliation: Comparative Orthopedic Molecular Medicine Laboratory, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, 43210, USA.
authors.forename: Akikazu
authors.initials: A
authors.lastname: Ishihara
authors.forename: Terri A
authors.initials: TA
authors.lastname: Zachos
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Bartlett
authors.forename: Alicia L
authors.initials: AL
authors.lastname: Bertone
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D000818:Animals; D001854:Bone Marrow Cells; D019902:Chondrocytes / Q000166:cytology* / Q000821:virology; D015870:Gene Expression; D015316:Genetic Therapy / Q000009:adverse effects / Q000662:veterinary*; D005822:Genetic Vectors / Q000009:adverse effects; D006367:HeLa Cells; D006736:Horses / Q000821:virology; D006801:Humans; D013234:Stem Cells / Q000166:cytology* / Q000821:virology; D013583:Synovial Membrane / Q000166:cytology* / Q000821:virology