Previous studies indicate that immunization with recombinant (r) vaccinia-human immunodeficiency virus type 1 (HIV-1) gp160 and boosting with baculovirus-derived HIV-1 rgp160 results in stronger cellular and antibody responses than those following either vaccine alone. The durability of immunity over 1 year was evaluated in 12 recipients. Both cellular and binding antibody responses remained detectable but diminished, and neutralizing antibodies were absent. To boost immunity, rgp160 was given again 1 year after the initial boost. Reboosting elicited strong HIV-specific lymphoproliferative responses. Binding antibody levels also rose dramatically, and the magnitude of the peak responses was significantly greater following the 2-year than following the 1-year boost. However, neutralizing antibody titers were low (1:10-1:20) and detected in only 4 of 12 persons. Moreover, persistent CD8+ cytolytic responses were not induced. Thus, although repeated rgp160 boosting after vaccinia-envelope priming can augment selected immune components, an altered regimen may be necessary to achieve protective long-term immunity to HIV-1.