Adenovirus-mediated gene transfer to the ocular surface epithelium

Exp Eye Res. 1998 Nov;67(5):531-8. doi: 10.1006/exer.1998.0557.


Gene transfer to the ocular surface epithelium is of potential therapeutic value. It was determined whether a reporter gene can be introduced into the ocular surface epithelium in vitro (human cell lines), ex vivo (human tissues), and in vivo (rats) by treating with a recombinant, replication-deficient, adenovirus type 5. Human and conjunctival cell lines were cultured with various multiplicities of infection (MOI; 3.2x10(-5)-5x10(-1)) of adenovirus vector (Ad5:Adex1CAlacZ) containing the reporter gene lacZ (1.3-2.0x10(4) PFU ml-1). The ex vivo study used human corneal and conjunctival tissues obtained from an eye bank and during surgery. Non-specific upregulation of inflammatory cytokines of conjunctival epithelium infected by Ad5 was assayed and its suppression by steroids. For the in vivo study, Ad5 (5x10(5) PFU, 5-10 microliter) was applied to the eyes of 8-12-week-old cotton rats, which were enucleated 24 and 48 hr later. The maximum lacZ expression in vitro was demonstrated in the corneal epithelial cell line at 7 days (1x10(-1) MOI) and conjunctival epithelial cell line at 2 days (4x10(-4) MOI). Furthermore, lacZ was also expressed in the superficial corneal and conjunctival epithelium in the ex vivo study. IL-6, IL-8, and ICAM-1 expression from conjunctival epithelium by Ad5 was significantly inhibited by treatment with betamethasone (BM). For the in vivo study, only the conjunctival epithelium demonstrated beta-Gal activity at 24 and 48 hr after application. These data indicate that adenovirus vector is capable of directly delivering gene to the corneal and conjunctival epithelium, suggesting a variety of possible gene therapy uses. The concomitant application of steroid eye drops may avoid inflammation.

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Anti-Inflammatory Agents / pharmacology
  • Betamethasone / pharmacology
  • Cell Culture Techniques
  • Cell Line
  • Conjunctiva / cytology*
  • Conjunctiva / metabolism
  • Epithelium, Corneal / cytology*
  • Epithelium, Corneal / metabolism
  • Gene Transfer Techniques*
  • Genes, Reporter
  • Genetic Vectors*
  • Humans
  • Interleukin-6 / biosynthesis
  • Interleukin-8 / biosynthesis
  • Lac Operon
  • Sigmodontinae
  • Up-Regulation / drug effects
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism


  • Anti-Inflammatory Agents
  • Interleukin-6
  • Interleukin-8
  • Betamethasone
  • beta-Galactosidase