Abstract
Abstract: :
Purpose: Feline immunodeficiency virus (FIV)-based vectors effectively transduce a lacZ reporter gene into retinal ganglion cells (RGCs) in culture, resulting in a high transduction efficiency. The purpose of this study was to evaluate FIV vector mediated gene transfer to the retina in vivo. Methods: FIV vector mediated gene transfer to the retina in vivo was evaluated in adult Sprague Dawley rats using intravitreal and subretinal injections of FIV vector containing the lacZ reporter gene. One group of rats received an intravitreal injection of 6µl of FIV vector through the pars plana of one eye. The second group of rats received an intravitreal injection of 6µl of FIV vector through the pars plana of one eye, with additional scraping of the internal limiting membrane. The third group of rats received a subretinal injection of FIV vector. The titer of the lentiviral vector was 5 x 108 TU/ml. The fellow eye was not injected and served as the control. X-gal staining was used to determine lacZ expression at 96 hours. Results: Eyes injected subretinally with FIV vector showed blue X-gal staining after 96 hours. Retinal cross sections showed high efficiency transduction of the retinal pigment epithelium (RPE). No lacZ expression was present in the fellow (control) eyes. Eyes injected intravitreally did not show lacZ expression in the retinal ganglion cells (RGCs). Removal of the internal limiting membrane in eyes injected intravitreally did not affect lacZ expression. Conclusions: In the adult rat, subretinal injections of FIV vector result in significant transduction of the RPE. However intravitreal injections of FIV vector using the same viral titer do not result in transduction of the RGCs.
Keywords: neuroprotection • gene transfer/gene therapy • ganglion cells