Fifty-three (25%) of 215 keratectomy specimens of patients with herpes simplex stromal keratitis displayed granulomatous reactions at the level of Descemet's membrane (50/53), midstroma (13/53), and Bowman's layer (5/53). Using an immunoperoxidase technique, herpes simplex virus (HSV) antigens were detected in keratocytes, endothelial cells, and foci of epithelioid histiocytes and multinucleated giant cells around Descemet's membrane. Both granulomatous reactions and HSV antigens were identified significantly more often in specimens with ulcerative necrotizing stromal keratitis than in those from patients with stromal scarring or nonulcerative nonnecrotizing keratitis (P less than 0.00001 and P less than 0.005, respectively). Herpes simplex virus antigens also were present in endothelial cells adjacent to foci of granulomatous reactions around Descemet's membrane in association with disciform stromal scarring. To our knowledge, this is the first demonstration of HSV antigens in human corneal endothelial cells and in the granulomatous reactions at the level of Descemet's membrane.