Macrophage migration inhibition factor (MIF), a soluble mediator of delayed hypersensitivity, was assayed for in the aqueous humor of rabbits undergoing corneal graft rejection. Penetrating and interlamellar xenografts and interlamellar allografts were performed in rabbits, and aqueous humor was obtained during the rejection. Significant levels of MIF activity were measured in the aqueous humor early in the course of xenograft and allograft rejection, and MIF activity was present during the course of the active rejection. This activity returned to near normal after the active rejection resolved. No significant MIF activity could be measured during the nonspecific inflammations produced by alkali burns, multiple paracenteses, intracorneal clove oil, or mechanical debriding of the endothelium.