Adenovirus-mediated gene transfer to human lens epithelial cells in organ culture

Purpose: To assess the feasibility of using recombinant adenovirus vectors to transduce the human lens epithelial cells (LECs) involved in posterior c apsule opacification (PCO). Setting: Department of Ophthalmology and Molecular Medicine Unit, Universit y of Manchester, Manchester, United Kingdom. Methods: Seventeen human lens capsules were maintained in organ culture to allow LECs to proliferate onto the posterior capsule. Partly covered and co mpletely covered capsules were infected with a recombinant adenovirus vecto r RAd35, encoding for the marker gene beta-galactosidase at plaque-forming units per milliliter (pfu/mL) ranging from 10(7) to 10(10) for up to 48 hou rs. Assessment of infection and transduction of the marker gene were achiev ed by calculating the percentage of cells exhibiting X-gal staining both ma croscopically and microscopically. Results: Staining appeared to be dependent on virus dose, with most intense staining at doses of 10(8) and 10(9) pfu/ml with decreased staining at hig her and lower viral doses. Microscopic assessment demonstrated that all cel ls expressed beta-galactosidase when infected with 10(9) piu, 84% at 10(8) pfu, and 45% at 10(7) pfu. At 10(10) pfu, some cytotoxicity was observed. Conclusions: These results indicate that recombinant adenoviruses can be us ed to transfer genes to the LECs involved in PCO. The transfer of cytotoxic genes after cataract surgery may be considered a preventive measure for PC O. (C) 2000 ASCRS and ESCRS.