Tears, conjunctival epithelium, and corneoscleral tissue from AIDS patients were used for the isolation of HTLV-III and also for identifying cell types which support its replication. HTLV-III was isolated from tears of AIDS patients (66.6%) by cultivation of cells and fluid from patients' eyes with fresh human mononuclear cells. The cells from the conjunctival scrapings of these patients (33.3%) revealed HTLV-III antigens by indirect immunofluorescence (IF) using anti-P24 and P15 monoclonal antibodies. HTLV-III from the cell-free supernatant of the infected mononuclear cells from two patients' cocultures were further transmitted into fresh cells. The cells from right and left central cornea, as well as limbal cornea from an asymptomatic HTLV-III antibody-positive individual and one AIDS patient revealed HTLV-III upon cocultivation. HTLV-III P15 and P24 antigens were detected in cultured primary cornea epithelial cells. The tears and conjunctival cells from a control group were found to be free of HTLV-III. Although no documented cases of AIDS have been reported in corneal transplant recipients, serologic screening of donors prior to the use of the tissues for transplantation is advisable. Our data also raises important questions regarding possible transmission of virus during ophthalmologic examination by way of examiner's hands, through instruments and during contact lens fittings. Moreover, these findings indicate the need for testing various eye disinfectants for virus inactivation and/or inhibition.