We examined the effect of a synthetic oligodeoxynucleotide containing unmethylated CpG immunostimulatory sequences (ISS) as an adjuvant for an HIV-1 immunogen (inactivated, gp120-depleted HIV-1 virus particles). The addition of the ISS to HIV-1 in incomplete Freund's adjuvant (IFA) was the optimal combination for the production of HIV-1-specific immune responses as measured by IFN-gamma (p=0.002) and IgG antibody production. Furthermore, the group that was immunized with HIV/IFA/ISS, as well as the group that received HIV-1 in complete Freund's adjuvant (CFA), stimulated significantly more RANTES (a beta-chemokine) (p=0.002) production from lymph node cells. These results suggest that the addition of CpG immunostimulatory sequences to HIV antigens in IFA may optimize HIV-1-specific immune responses and provides further rationale for the testing of ISS in combination with gp120-depleted whole-killed HIV-1 in IFA as a potential preventive or therapeutic HIV-1 vaccine.