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

J Cataract Refract Surg. 2000 Jun;26(6):887-92. doi: 10.1016/s0886-3350(00)00325-4.

Abstract

Purpose: To assess the feasibility of using recombinant adenovirus vectors to transduce the human lens epithelial cells (LECs) involved in posterior capsule opacification (PCO).

Setting: 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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Cell Division
  • Epithelial Cells / metabolism*
  • Epithelial Cells / virology
  • Feasibility Studies
  • Gene Transfer Techniques*
  • Genetic Vectors
  • Humans
  • Lens Capsule, Crystalline / metabolism*
  • Lens Capsule, Crystalline / virology
  • Organ Culture Techniques
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • beta-Galactosidase