Herpes simplex virus mediated gene transfer to primate ocular tissues

Xuyang Liu, Curtis R. Brandt, B'Ann T. Gabelt, Paul J. Bryar, Morton E. Smith, Paul L. Kaufman*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

56 Scopus citations

Abstract

We evaluated the feasibility of delivering a gene into monkey eyes using a replication-competent herpes simplex virus (HSV) type 1 ribonucleotide reductase mutant (hrR3) expressing the Escherichia coli lacZ gene. To determine the efficiency of in vitro HSV-mediated gene transfer, cultured human trabecular meshwork (HTM) and human ciliary muscle (HCM) cells were infected with hrR3 and β-galactosidase activity was measured histochemically. Six cynomolgus monkey eyes received viral injections into the anterior chamber (2 x 107 pfu) and/or the vitreous (5 x 107 pfu), and the distribution of cells expressing lacZ was evaluated. In vitro, both cultured HTM and HCM cells displayed multiplicity-dependent β-galactosidase activity. In vivo, intracameral and/or intravitreal injection resulted in transgene expression in TM cells and in non-pigmented ciliary epithelial cells (NPE), but not in CM cells. Transgene expression was also detected in retinal pigmented epithelial (RPE) cells and sporadic retinal ganglion cells (RGC) in eyes receiving virus intracamerally and intravitreally respectively. We observed significant inflammation in the anterior chamber, TM and CM in virus-injected eyes, along with mild vitritis and retinitis. This study demonstrates successful gene transfer using hrR3 as a vector in human ocular cells and in ocular tissues in living monkeys. Further investigation of the etiology of the inflammatory response, possible cytotoxicity, and limited duration of transgene expression is necessary in order to make this technique clinically applicable.

Original languageEnglish (US)
Pages (from-to)385-395
Number of pages11
JournalExperimental eye research
Volume69
Issue number4
DOIs
StatePublished - Oct 1999

Keywords

  • Ciliary epithelium
  • Ciliary muscle
  • Cynomolgus monkey
  • Gene therapy
  • Gene transfer
  • Herpes simplex virus type 1
  • LacZ
  • Retinal ganglion cells
  • Ribonucleotide reductase mutant
  • Trabecular meshwork
  • β- galactosidase

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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