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

Louis M. Carrington, Tom Southgate, Lisa A. Saxby, Khaled Abul-Hassan, Tricia C. Maleniak, Maria G. Castro, Michael E. Boulton

Research output: Contribution to journalArticle

8 Citations (Scopus)

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 β-galactosidase at plaque-forming units per milliliter (pfu/mL) ranging from 107 to 1010 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 108 and 109 pfu/mL with decreased staining at higher and lower viral doses. Microscopic assessment demonstrated that all cells expressed β-galactosidase when infected with 109 pfu, 84% at 108 pfu, and 45% at 107 pfu. At 1010 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. (C) 2000 ASCRS and ESCRS.

Original languageEnglish (US)
Pages (from-to)887-892
Number of pages6
JournalJournal of Cataract and Refractive Surgery
Volume26
Issue number6
DOIs
StatePublished - Jun 2000
Externally publishedYes

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Organ Culture Techniques
Capsule Opacification
Adenoviridae
Lenses
Epithelial Cells
Galactosidases
Staining and Labeling
Capsules
Genes
Molecular Medicine
Ophthalmology
Cataract
Viruses
Infection

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Carrington, L. M., Southgate, T., Saxby, L. A., Abul-Hassan, K., Maleniak, T. C., Castro, M. G., & Boulton, M. E. (2000). Adenovirus-mediated gene transfer to human lens epithelial cells in organ culture. Journal of Cataract and Refractive Surgery, 26(6), 887-892. https://doi.org/10.1016/S0886-3350(00)00325-4

Adenovirus-mediated gene transfer to human lens epithelial cells in organ culture. / Carrington, Louis M.; Southgate, Tom; Saxby, Lisa A.; Abul-Hassan, Khaled; Maleniak, Tricia C.; Castro, Maria G.; Boulton, Michael E.

In: Journal of Cataract and Refractive Surgery, Vol. 26, No. 6, 06.2000, p. 887-892.

Research output: Contribution to journalArticle

Carrington, LM, Southgate, T, Saxby, LA, Abul-Hassan, K, Maleniak, TC, Castro, MG & Boulton, ME 2000, 'Adenovirus-mediated gene transfer to human lens epithelial cells in organ culture', Journal of Cataract and Refractive Surgery, vol. 26, no. 6, pp. 887-892. https://doi.org/10.1016/S0886-3350(00)00325-4
Carrington, Louis M. ; Southgate, Tom ; Saxby, Lisa A. ; Abul-Hassan, Khaled ; Maleniak, Tricia C. ; Castro, Maria G. ; Boulton, Michael E. / Adenovirus-mediated gene transfer to human lens epithelial cells in organ culture. In: Journal of Cataract and Refractive Surgery. 2000 ; Vol. 26, No. 6. pp. 887-892.
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