Molecular analysis of acyclovir-induced suppression of HSV-1 replication in rabbit corneal cells.
An in vitro HSV drug-suppression model has been established in rabbit corneal cell (SIRC) monolayers. Confluent SIRC monolayers were inoculated with McKrae strain HSV-1 (0.014 PFU/cell) and subsequently suppressed with acyclovir (ACV) (40 micrograms/ml) after adsorption for 1 hr at 37 degrees C. On days 0-5 postinoculation (PI), infectious HSV-1 was detected in approximately 50% of the SIRC cell cultures by standard supernatant and cell-free homogenate co-culture assays. On days 6-10 postsuppression, infectious HSV was detected in only 7-19% of the cultures. After removal of ACV suppression in the cultures (day 11 PI), HSV cytopathology developed to a 3-4+ level within 2-5 days. Dot blot hybridization of DNA extracted from the cultures indicated that the HSV genome was retained consistently in SIRC cells at a level of 0.0015-0.015 copies per cell during active ACV-suppression. During ACV-suppressed infection, approximately 0.9% of the SIRC cells contained the HSV genome as demonstrated by blot hybridization. After removal of ACV, a 1000-fold increase in HSV DNA concentration in SIRC cell cultures was detected.
Rong
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BL
Eye Research Institute, Boston, Massachusetts 02114.
Dunkel
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EC
Pavan-Langston
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D000212:Acyclovir / Q000494:pharmacology*; D000818:Animals; D003315:Cornea / Q000166:cytology / Q000382:microbiology*; D004279:DNA, Viral; D009693:Nucleic Acid Hybridization; D011817:Rabbits; D018139:Simplexvirus / Q000502:physiology*; D014779:Virus Replication / Q000187:drug effects*
D004279:DNA, Viral; D000212:Acyclovir