Abstract
Objective - To determine whether the active metabolite of leflunomide, A77 1726 (A77), inhibits replication of feline herpesvirus-1 (FHV-1) in cell culture. Study Population - Crandell Rees feline kidney (CRFK) cell cultures. Procedures - Cell cultures were inoculated with FHV-1 and treated simultaneously with concentrations of A77 ranging from 0 to 200 μM. The antiviral effect of A77 was determined by use of conventional plaque reduction assays. The effect of A77 on viral load was determined via real-time PCR analysis, and transmission electron microscopy was used to evaluate the effect of A77 on viral morphology. To determine whether the antiviral effect was attributable to alterations in CRFK cell viability and number, CRFK cells were treated with various concentrations of A77 and stained with Annexin V and propidium iodide to assess apoptosis and a mitochondrial function assay was used to determine cell viability. Results - Concentrations of A77 ≥ 20μM were associated with substantial reduction in plaque number and viral load. Concentrations ≥ 100μM were associated with complete suppression of plaque formation. At low concentrations of A77, clusters of intracytoplasmic virus particles that appeared to lack tegument and an external membrane were detected. Treatment of uninfected CRFK cell monolayers with A77 was associated with reduction in mitochondrial function with minimal evidence of apoptosis. Conclusions and Clinical Relevance - Leflunomide may be an alternative to current calcineurin-based immunosuppressive protocols used in feline organ transplantation because of its antiherpesviral activity.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1010-1015 |
| Number of pages | 6 |
| Journal | American Journal of Veterinary Research |
| Volume | 68 |
| Issue number | 9 |
| DOIs | |
| State | Published - Sep 2007 |
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- veterinary(all)
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In vitro effects of the active metabolite of leflunomide, A77 1726, on feline herpesvirus-1. / Williams, Courtney R.; Sykes, Jane E; Mehl, Margo; MacLeod, Jennifer S.; Lindsay, LeAnn L.; Poland, Amy M.; Chen, Yi-Je; Kyles, Andrew E.; Waldman, W. James; Gregory, Clare R.
In: American Journal of Veterinary Research, Vol. 68, No. 9, 09.2007, p. 1010-1015.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - In vitro effects of the active metabolite of leflunomide, A77 1726, on feline herpesvirus-1
AU - Williams, Courtney R.
AU - Sykes, Jane E
AU - Mehl, Margo
AU - MacLeod, Jennifer S.
AU - Lindsay, LeAnn L.
AU - Poland, Amy M.
AU - Chen, Yi-Je
AU - Kyles, Andrew E.
AU - Waldman, W. James
AU - Gregory, Clare R.
PY - 2007/9
Y1 - 2007/9
N2 - Objective - To determine whether the active metabolite of leflunomide, A77 1726 (A77), inhibits replication of feline herpesvirus-1 (FHV-1) in cell culture. Study Population - Crandell Rees feline kidney (CRFK) cell cultures. Procedures - Cell cultures were inoculated with FHV-1 and treated simultaneously with concentrations of A77 ranging from 0 to 200 μM. The antiviral effect of A77 was determined by use of conventional plaque reduction assays. The effect of A77 on viral load was determined via real-time PCR analysis, and transmission electron microscopy was used to evaluate the effect of A77 on viral morphology. To determine whether the antiviral effect was attributable to alterations in CRFK cell viability and number, CRFK cells were treated with various concentrations of A77 and stained with Annexin V and propidium iodide to assess apoptosis and a mitochondrial function assay was used to determine cell viability. Results - Concentrations of A77 ≥ 20μM were associated with substantial reduction in plaque number and viral load. Concentrations ≥ 100μM were associated with complete suppression of plaque formation. At low concentrations of A77, clusters of intracytoplasmic virus particles that appeared to lack tegument and an external membrane were detected. Treatment of uninfected CRFK cell monolayers with A77 was associated with reduction in mitochondrial function with minimal evidence of apoptosis. Conclusions and Clinical Relevance - Leflunomide may be an alternative to current calcineurin-based immunosuppressive protocols used in feline organ transplantation because of its antiherpesviral activity.
AB - Objective - To determine whether the active metabolite of leflunomide, A77 1726 (A77), inhibits replication of feline herpesvirus-1 (FHV-1) in cell culture. Study Population - Crandell Rees feline kidney (CRFK) cell cultures. Procedures - Cell cultures were inoculated with FHV-1 and treated simultaneously with concentrations of A77 ranging from 0 to 200 μM. The antiviral effect of A77 was determined by use of conventional plaque reduction assays. The effect of A77 on viral load was determined via real-time PCR analysis, and transmission electron microscopy was used to evaluate the effect of A77 on viral morphology. To determine whether the antiviral effect was attributable to alterations in CRFK cell viability and number, CRFK cells were treated with various concentrations of A77 and stained with Annexin V and propidium iodide to assess apoptosis and a mitochondrial function assay was used to determine cell viability. Results - Concentrations of A77 ≥ 20μM were associated with substantial reduction in plaque number and viral load. Concentrations ≥ 100μM were associated with complete suppression of plaque formation. At low concentrations of A77, clusters of intracytoplasmic virus particles that appeared to lack tegument and an external membrane were detected. Treatment of uninfected CRFK cell monolayers with A77 was associated with reduction in mitochondrial function with minimal evidence of apoptosis. Conclusions and Clinical Relevance - Leflunomide may be an alternative to current calcineurin-based immunosuppressive protocols used in feline organ transplantation because of its antiherpesviral activity.
UR - http://www.scopus.com/inward/record.url?scp=34548634388&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34548634388&partnerID=8YFLogxK
U2 - 10.2460/ajvr.68.9.1010
DO - 10.2460/ajvr.68.9.1010
M3 - Article
C2 - 17764417
AN - SCOPUS:34548634388
VL - 68
SP - 1010
EP - 1015
JO - American Journal of Veterinary Research
JF - American Journal of Veterinary Research
SN - 0002-9645
IS - 9
ER -