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DA Leske, N Loewen, Y Chen, MP Fautsch, EM Poeschla, JM Holmes; Comparison of FIV and Adenovirus Mediated Long Term Transgene Expression in the Neonatal Rat Retina . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4611.
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Purpose: To compare the long-term expression of a transgene (lacZ) delivered to neonatal rat retina by either feline immunodeficiency virus (FIV) or adenovirus (AdV) vectors. Replication-deficient FIV vectors are derived from a genomically integrating lenti-retrovirus whereas AdV remains episomal. These factors may influence persistence. The neonatal rat retina was chosen because of its use in models for retinal neovascularization, particularly for retinopathy of prematurity (ROP) and diabetic retinopathy. Methods: The right eyes of 44 five day old Sprague Dawley rats received subretinal injections of FIV2.CMV.lacZ, a VSV-G pseudotyped lacZ-encoding FIV vector (2 µl of 1 X 105 TDU/µl). Left eyes were injected subretinally with AdV vector containing the lacZ gene (2 µl of 1 X 105 TDU/µl). Animals were sacrificed at 2 days (n=14), 1 week (n=10), 4 weeks (n=10) and 12 weeks (n=10) after injection. Eyes were fixed, cornea and lens removed and eyecups stained for lacZ expression. Eyecups were graded in a masked manner for expression (colorimetric assay; incidence and extent, based on a scale from 0 to 4) along with non-injected controls. Incidence and extent were compared between FIV and AdV (Chi-square and Wilcoxon signed-rank tests). Representative eyecups were embedded in paraffin and sectioned for localization of lacZ expression. Results: Twelve weeks post-injection, the extent of transduction was greater in FIV injected eyes than AdV injected eyes (median grade 2.5 vs 1.5, p=0.03). 40% of eyes injected with FIV showed total retinal transduction at 12 weeks vs. 0% of eyes injected with AdV. Overall, 10 of 10 FIV injected eyes showed some retinal transduction at 12 weeks, compared to 9 of 10 AdV injected eyes. There were no significant differences in incidence or extent of transduction at the 2 day, 1 week and 4 week timepoints (incidence range 80% - 100%, severity median range 1 to 2). LacZ expression was localized primarily to the RPE in both FIV and AdV injected eyes and there was preservation of retinal architecture. In addition to RPE, both vectors showed variable lacZ expression in inner retinal layers at 4 weeks and 12 weeks. Conclusion: Gene transfer using an FIV vector resulted in greater extent of retinal transduction than AdV at 12 weeks following subretinal injection. FIV vectors show more promise than AdV as long-term delivery systems for in vivo gene transfer into retina and may have specific clinical applications to neovascular retinal diseases such as ROP, diabetic retinopathy, and age-related macular degeneration.
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