title: Recombinant AAV-transduced iris pigment epithelial cell transplantation may transfer vector to native RPE but suppress systemic dissemination.
abstract: OBJECTIVE
  To determine whether adenoassociated virus (AAV) vectors transduced into iris pigment epithelial (IPE) cells and transplanted into the subretinal space of rats will transfer the AAV genome to the host cells and whether the vectors are disseminated systemically.
  
  
  METHODS
  Recombinant (r)AAV was transduced into rat IPE cells and transplanted into the subretinal space of rats. For the control, rAAVs alone were injected subretinally. The transplanted IPE cells were detected by LacZ staining. Immunohistochemistry, electron microscopy, electroretinography, and fluorescein-dextran angiography were performed. DNA was extracted from various organs and blood and examined for the AAV genome by polymerase chain reaction.
  
  
  RESULTS
  No toxicity from rAAV transduction was observed in vitro. LacZ was expressed in the transplanted cells 1 and 2 weeks after transplantation. At 4 and 12 weeks, fewer transplanted cells were detected than at 1 week, and LacZ expression was occasionally detected at the level of host retinal pigment epithelial (RPE) cells. Expression was also detected in ciliary body epithelial cells. The electroretinograms and fluorescein-dextran angiography were only mildly altered. Significantly lower levels of AAV genome were detected in the organs and blood of rats receiving rAAV-IPE cell transplants than with direct intravenous injection of AAV vectors.
  
  
  CONCLUSIONS
  AAV-mediated LacZ was expressed in the transplanted cells after subretinal transplantation, and the transplanted IPE cells may transfer the rAAV to host tissues, such as RPE cells, long after the transplantation. This method of gene delivery did not lead to systemic dissemination of the vectors.
authors.affiliation: Department of Ophthalmology and Visual Science, Tohoku University Graduate School of Medicine, Sendai, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yoshioka
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Abe
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Wakusawa
authors.forename: Takuya
authors.initials: T
authors.lastname: Moriya
authors.forename: Shizue
authors.initials: S
authors.lastname: Mochizuki
authors.forename: Yoko
authors.initials: Y
authors.lastname: Saigo
authors.forename: Takae
authors.initials: T
authors.lastname: Saito
authors.forename: Hiromi
authors.initials: H
authors.lastname: Murata
authors.forename: Yumi
authors.initials: Y
authors.lastname: Tokita
authors.forename: Takashi
authors.initials: T
authors.lastname: Iseya
authors.forename: Eriko
authors.initials: E
authors.lastname: Sugano
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tomita
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Sagara
authors.forename: Makoto
authors.initials: M
authors.lastname: Tamai
mesh_terms: D000818:Animals; D002470:Cell Survival; D017690:Cell Transplantation; D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis; D000229:Dependovirus / Q000235:genetics*; D003911:Dextrans; D004596:Electroretinography; D005110:Extracellular Space; D005452:Fluoresceins; D005822:Genetic Vectors; D016679:Genome, Viral; D007150:Immunohistochemistry; D007498:Iris / Q000166:cytology*; D007763:Lac Operon / Q000502:physiology; D008856:Microscopy, Fluorescence; D010857:Pigment Epithelium of Eye / Q000637:transplantation* / Q000821:virology*; D016133:Polymerase Chain Reaction; D051381:Rats; D020318:Rats, Long-Evans; D012160:Retina / Q000601:surgery* / Q000821:virology; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D003911:Dextrans; D005452:Fluoresceins; C052526:fluorescein-dextran; D001616:beta-Galactosidase