The results of this investigation demonstrate that the IgA antibody in tears inhibits the attachment of herpes simplex virus (HSV) to cells and implicate immunoglobulin A (IgA) isotype antibody in the protection of the ocular surface. Tears were collected from the eyes of infected and uninfected rabbits and tested for the presence of infectious virus, for viral neutralization titer, and in an enzyme-linked immunoassay (ELISA) for antibody to HSV. Neither the tears from the uninfected rabbits nor the tears from the infected rabbits produced viral cytopathic effects on indicator cells in vitro. Tears from uninfected rabbits had no virus neutralization capacity while the tears from infected rabbits had titers of 1:10 to 1:40. Tears collected from HSV-1 infected rabbits, but not uninfected rabbits, had anti-HSV antibody titers as determined by the enzyme-linked immunosorbent assay. Lectin affinity column enrichment of IgA from the tears of uninfected and infected rabbits was examined for the capacity to block viral adsorption to cells using32P-labeled virus. It was found that exposure of the radio-actively-labeled virus to the purified IgA fraction of tears from infected rabbits reduced the amount of virus binding to Vero cells, whereas the purified IgA obtained from uninfected tears had minimal activity. These results may imply a role for the IgA-containing fraction of tears from HSV-infected animals in preventing viral attachment to cells and suggest that antibody of this isotype plays a protective role at the ocular surface.