Because of the importance of the envelope glycoprotein (Env) in determining the pathogenicity of HIV-1 and the importance of the immune response to Env in controlling virus spread, attempts are being made to study HIV-1 Env-directed immunity in primate models. To date HIV-1 Env-specific effector T lymphocyte responses have not been demonstrated in virus-infected nonhuman primates. We have previously reported that cynomolgus monkeys can develop a persistent infection with a chimeric simian-human immunodeficiency virus (SHIV) composed of SIVmac239 carrying the HIV-1 env, tat, rev, and vpu genes. We now demonstrate that SHIV-infection of another macaque species, the rhesus monkey, generates persistent, HIV-1 Env-specific cytolytic T lymphocyte (CTL) responses. These CTL are CD8+ and major histocompatibility complex (MHC) class I-restricted. The induction of CTL was correlated neither to the virus load nor to the MHC class I haplotypes of the monkeys. The SHIV-infected rhesus monkey can, therefore, now be employed for studying effector T lymphocyte recognition of HIV-1 Env.