OBJECTIVE To determine whether the subretinal space supports the induction of deviant immune responses to cell-associated and soluble antigens and to elucidate factors influencing the immunologic properties of the subretinal space. METHODS P815 mastocytoma cells were used as cell-associated antigens and were inoculated into the anterior chamber (AC), the vitreous cavity (VC), the subretinal space, and subconjunctivally in H2-compatible, but minor H-incompatible, BALB/c mice. Ovalbumin, as a soluble antigen, was similarly injected into eyes, after which recipient animals were immunized with ovalbumin and complete Freund's adjuvant. Delayed-type hypersensitivity (DTH) was assessed by ear challenge. To alter the conditions in the subretinal space, the outer blood-retinal barrier was disrupted by compromising retinal pigment epithelial (RPE) cells with a systemic injection of sodium iodate. Immune privilege in the AC was abolished by mild corneal cauterization. RESULTS Antigen-specific DTH did not develop in mice in which alloantigenic tumor cells or ovalbumin was injected into the AC, the VC, or the subretinal space, and the mice's spleens contained lymphocytes capable of suppressing DTH when adoptively transferred into naive mice. When RPE cells were compromised with sodium iodate, tumor cells or ovalbumin injected into the subretinal space or the VC did not induce immune deviation, although the AC of these eyes still promoted AC-associated immune deviation. By contrast, when immune privilege in the AC was abolished by corneal cauterization, neither tumor cells nor ovalbumin injected into the subretinal space or the VC of eyes elicited immune deviation. CONCLUSIONS The subretinal space supports immune deviation for histoincompatible tumor cells and soluble protein antigens by actively suppressing antigen-specific DTH. Acute loss of immune privilege in the subretinal space and the VC does not cause loss of privilege in the AC, but abolition of immune privilege in the AC eliminates the capacity of the subretinal space and the VC to support immune deviation to antigens injected locally.