Topical treatment of primary herpetic keratitis with 0.25% retinoic acid was effective in significantly diminishing the severity of epithelial lesions in rabbits. Antiviral activity against herpes simplex virus type 1 was also demonstrated in cell culture. Retinoic acid treatment of virus particles for 24 hours had no effect on the infectivity of the virus. At effective antiviral concentrations in cell culture, retinoic acid had toxic action on two different cell lines as detected by an increase in cellular generation time or inhibition of DNA synthesis. Topical instillation of retinoic acid into the eyes of rabbits at the effective antiviral dose (0.25%) had no inhibitory effect on corneal wound closure but did reduce the rate of incorporation of thymidine into DNA by 27%. These studies suggest that although retinoic acid may inhibit the replication of herpes simplex virus type 1, it is difficult to distinguish antiviral effects from anticellular effects.