We have evaluated alum, immunostimulatory cytosine guanine dinucleotide-containing oligodeoxy-nucleotides (CPG ODN), and an alum/CPG ODN combination as adjuvants with hepatitis B surface antigen, to compare their potential to allow lower doses of antigen to be used for induction of humoral responses. BALB/c mice were immunized by intramuscular injection with 0.01, 0.1 or 1.0 micro g recombinant hepatitis B surface antigen without adjuvant or with alum and/or CPG ODN added. When given without adjuvant or with alum, each 10-fold decrease in amount of antigen resulted in a similarly reduced titre of antibody against hepatitis B surface antigen. In contrast, CPG ODN, on its own or combined with alum, allowed high anti-hepatitis B surface antigen titres even with a 1000-fold reduction in amount of antigen. These findings may have important immunological and economical consequences for vaccine development.