Adenovirus-mediated gene transfer to corneal and conjunctival epithelium by topical application

Kazuo Tsubota, M. Ono, H. Ando, H. Inoue, I. Saito

Research output: Contribution to journalArticle

Abstract

Purpose. Gene transfer to the ocular surface epithelium holds the potential for therapy and delivery of therapeutic proteins to the surface epithelium. We determined whether a reporter gene can be introduced into ocular surface epithelium in vitro, ex vivo and in vivo using a recombinant replication-deficient adenovirus. Methods. Adenovirus vector containing the reporter gene of lacZ (1.3 to 2.0X104 PFU) was added to the culture medium of human corneal and conjunctival cell lines. The ex vivo study used human corneal and conjunctival tissues obtained in surgery. In vivo study, adenovirus vector (5 X 105 PFU, 1.5-3.0 μl) was applied topically to the eyes of 7 week-old cotton rats (all female, N=14), which were enucleated at 24 and 48 hours after application. Results. The lacZ expression was demonstrated in corneal epithelium cell line at day 2 (8.0X102 or more PFU) and conjunctival cell line (8.0X102 or less PFU) until day 7, as well as ex vivo study. Conjunctival epithelium showed the cell toxity with 4.0X103 or more PFU, however no side effects were observed in corneal epithelium. In vivo study, only the conjunctival epithelium was positive for β-Gal activity at 24 and 48 hours after topical application. Conclusions. Adenovirus vectors are capable to direct delivery of genes to the corneal and conjunctival epithelium with different optimal concentration without toxicity, suggesting a variety of possible gene therapy application.

Original languageEnglish
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
Publication statusPublished - 1996 Feb 15
Externally publishedYes

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Corneal Epithelium
Adenoviridae
Epithelium
Genes
Reporter Genes
Cell Line
Sigmodontinae
Genetic Therapy
Culture Media
Membrane Proteins
Therapeutics

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Adenovirus-mediated gene transfer to corneal and conjunctival epithelium by topical application. / Tsubota, Kazuo; Ono, M.; Ando, H.; Inoue, H.; Saito, I.

In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 3, 15.02.1996.

Research output: Contribution to journalArticle

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AU - Ono, M.

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AU - Saito, I.

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