A highly complex network of coinhibitory and costimulatory receptors regulates the outcome of virus-specific CD8⁺ T-cell responses. Here, we report on the expression patterns of multiple inhibitory receptors on HIV-specific, cytomegalovirus-specific, and bulk CD8⁺ T-cell memory populations. In contrast to cytomegalovirus-specific CD8⁺ T cells, the majority of HIV-specific CD8⁺ T cells exhibited an immature phenotype and expressed Programmed Death-1, CD160 and 2B4 but not lymphocyte activation gene-3. Notably, before antiretroviral therapy, simultaneous expression of these negative regulators correlated strongly with both HIV load and impaired cytokine production. Suppression of HIV replication by antiretroviral therapy was associated with reduced surface expression of inhibitory molecules on HIV-specific CD8⁺ T cells. Furthermore, in vitro manipulation of Programmed Death-1 and 2B4 inhibitory pathways increased the proliferative capacity of HIV-specific CD8⁺ T cells. Thus, multiple coinhibitory receptors can affect the development of HIV-specific CD8⁺ T-cell responses and, by extension, represent potential targets for new immune-based interventions in HIV-infected persons.