Adenovirus-mediated gene transfer to human lens epithelial cells in organ culture.
OBJECTIVE
To assess the feasibility of using recombinant adenovirus vectors to transduce the human lens epithelial cells (LECs) involved in posterior capsule opacification (PCO).


METHODS
Department of Ophthalmology and Molecular Medicine Unit, University 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 completely covered capsules were infected with a recombinant adenovirus vector 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 hours. Assessment of infection and transduction of the marker gene were achieved by calculating the percentage of cells exhibiting X-gal staining both macroscopically 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 higher and lower viral doses. Microscopic assessment demonstrated that all cells expressed beta-galactosidase when infected with 10(9) pfu, 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 used to transfer genes to the LECs involved in PCO. The transfer of cytotoxic genes after cataract surgery may be considered a preventive measure for PCO.
Carrington
L M
LM
Department of Ophthalmology, University of Manchester, Manchester, UK.
Southgate
T
T
Saxby
L A
LA
Abul-Hassan
K
K
Maleniak
T C
TC
Castro
M G
MG
Boulton
M E
ME
D000256:Adenoviridae / Q000235:genetics*; D002455:Cell Division; D004847:Epithelial Cells / Q000378:metabolism* / Q000821:virology; D005240:Feasibility Studies; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007903:Lens Capsule, Crystalline / Q000378:metabolism* / Q000821:virology; D009924:Organ Culture Techniques; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001616:beta-Galactosidase