May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
The Effect of Anti–viral Compounds on Herpes Simplex–1 Infected Corneal Epithelial Cells
Author Affiliations & Notes
  • D.K. Yoo
    Ophthalmology, Northwestern University, Chicago, IL
  • P.J. Bryar
    Ophthalmology, Northwestern University, Chicago, IL
  • M. Wong
    Ophthalmology, Northwestern University, Chicago, IL
  • A. Muthialu
    Ophthalmology, Northwestern University, Chicago, IL
  • Footnotes
    Commercial Relationships  D.K. Yoo, None; P.J. Bryar, None; M. Wong, None; A. Muthialu, None.
  • Footnotes
    Support  Support Research to Prevent Blindness Inc. NY, NY; Illinois Society for the Prevention of Blindness
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 4941. doi:
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      D.K. Yoo, P.J. Bryar, M. Wong, A. Muthialu; The Effect of Anti–viral Compounds on Herpes Simplex–1 Infected Corneal Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4941.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose:A cell culture model was used to assess the effect of using topical formulations of famciclovir and valacyclovir on a human corneal epithelial cell model infected with herpes simplex virus–1 (HSV–I). Methods:Minimal Essential Media (MEM) was used as the vehicle solution to mix all the reagents as well as to grow the human corneal cells. Vero cells (ATCC) were used to propagate human HSV–1, KOS strain. Vero cells were grown in MEM with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, 55mg/ml sodium pyruvate, and 0.1mM non–essential amino acids. SV–40 transfected human corneal epithelial cells (HCEC) were obtained from the Dr. Christopher Murphy (University of Wisconsin School of Veterinary Medicine), initially a gift from Kaoru Sasaki (Toyonaka Municipal Hospital, Osaka, Japan). These cells were grown in MEM with 0.5% dimethyl sulfoxide, 10% FBS, and 40µg/ml gentamicin. HCECs were grown to 80% confluency, and incubated with HSV–1 at 107.5 pfu for 2 hours in serum–free MEM; this infectious media was replaced with serum containing no drug, valcyclovir, or famciclovir (IC50, 50% inhibition of cell growth, equivalent to 13.6 µg/ml for valacyclovir and 0.6 µg/ml for famciclovir). Cells were incubated for 16 hours and at this time, cytopathic effect (CPE) was noted with cell rounding and detachment. DNA was isolated with trizol (Gibco BRL), and used for PCR. PCR primers for HSV–1 were designed for glycoprotein D, unique to the HSV–1 genome; glyceraldehyde–3–phosphate dehydrogenase (GAPDH) primers were used as a control. Results: Cells were plated in equal densities, reaching 80% confluency. They were subsequently infected with HSV–1 and treated with one of three media; without anti–viral agents, with valacyclovir, or with famciclovir. Harvesting of DNA was done after 16 hours. Microscopic observation showed that compared to the control and famciclovir group, the valacyclovir group had a greater number cells and greater amount of DNA. DNA isolated from the untreated cells was quantified by spectroscopy and found to be 0.18 µg/µl for the untreated group, 0.22 µg/µl for the valacyclovir group, and 0.05 µg/µl for the famciclovir group. Conclusions: A topical formulation of valacyclovir has been shown to inhibit cytopathic effects in comparison to famciclovir and no reagent.

Keywords: herpes simplex virus • cornea: epithelium • antiviral drugs 
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