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  • Author or Editor: Charlotte B. Keller x
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Abstract

Objective—To isolate and characterize pure cultures of feline corneal epithelial cells and to assess the extent and nature of feline herpesvirus (FHV)-1 infection in these cells.

Sample Population—Healthy eyes from 23 recently euthanatized cats.

Procedure—Stroma and epithelium of the rostral portion of the cornea were surgically isolated, and epithelial cells were detached from the stroma by enzymatic incubation. Epithelial cells were cultured in hormone-supplemented media. Cells were passaged, and cytokeratin expression was assessed. Cells were then infected with FHV-1, and cytopathic effects were determined.

Results—Cell cultures were readily established from samples obtained from each eye and could be maintained through 6 passages. Cultured cells expressed cytokeratins 3 and 12 but not other cytokeratins. Infection with FHV-1 was rapid and caused widespread cytopathic effects.

Conclusions and Clinical Relevance—Feline corneal cells cultured in vitro during multiple passages maintain consistent morphologic characteristics and intermediate filament expression. They are susceptible to infection with FHV-1 and may provide a useful in vitro model for investigation of ocular drugs. (Am J Vet Res 2005;66:205–209)

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in American Journal of Veterinary Research

Abstract

Objective— To assess the effect of cidofovir on viability of feline corneal epithelial (FCE) cells, replication of feline herpesvirus (FHV)-1, and virus-induced cytopathic changes.

Sample Population—Healthy eyes from 14 recently euthanatized cats.

Procedure—Cidofovir at concentrations ranging from 0.05 to 0.000005 mg/mL was added to primary cultures of FCE cells, and cytopathic changes and effects on cell proliferation and cell viability were determined during the subsequent 48 hours. Efficacy of cidofovir (0.02 and 0.05 mg/mL) to prevent in vitro infection of FCE cells with FHV-1 was determined during 72 hours of culture by assessing viral cytopathic effects and viral titers.

Results—Cidofovir at concentrations of 0.05, 0.005, and 0.0005 mg/mL significantly reduced mean viable cell counts, and cidofovir at a concentration of 0.05 mg/mL significantly reduced the percentage viability of cultured FCE cells. Minimal cytopathic changes were observed at concentrations of 0.02 and 0.05 mg of cidofovir/mL. Cidofovir at concentrations of 0.05 and 0.02 mg/mL abrogated the cytopathic effects attributable to FHV-1 infection and reduced viral titers from ≥ 1014 TCID50/mL to ≤ 103.5 TCID50/mL.

Conclusions and Clinical Relevance—Cidofovir in vitro was highly efficacious against FHV-1 infection of a primary culture of FCE cells but had cytostatic effects on cultured cells. (Am J Vet Res 2005;66:217–222)

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in American Journal of Veterinary Research

Abstract

Objective—To assess the effect of interferon (IFN)-α on viability of feline corneal epithelial cells, replication of feline herpesvirus (FHV)-1, and virus-induced cytopathic changes.

Sample Population—Healthy eyes from 10 recently euthanatized cats.

Procedure—4 replicate primary cultures of feline corneal epithelial cells were grown after the addition of 102 to 106 IU of IFN-α/mL. Cultures were examined every 24 hours for evidence of cytotoxic changes. Viable cell counts and percentage of viable cells were determined 48 hours after initiation of culture. In a separate experiment, cultures of corneal cells were inoculated with FHV-1 and cultured for 72 hours with or without 105 IU of IFN-α/mL. The FHV-1–infected cultures were evaluated for viral-induced cytopathic effects, and viral titers were determined in samples of culture supernatant.

Results—Interferon-α did not have cytotoxic effects on corneal epithelial cells at concentrations ranging from 102 to 106 IU of IFN-α/mL. Interferon-α at a concentration of 105 IU/mL significantly reduced the cytopathic changes and FHV-1 titers.

Conclusions and Clinical Relevance—Lack of in vitro cytotoxic effects and efficacy against FHV-1 infection in primary cultures of feline corneal cells suggests that the in vivo therapeutic effect of IFN-α should be assessed in controlled clinical trials. (Am J Vet Res 2005;66:210–216)

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in American Journal of Veterinary Research