The aim of this study was to compare the immunogenicity and antigenicity of cyclic and linear peptides that mimic the disulfide loops in HIV-2ROD gp125. Based on the hypothetical assignment of intrachain disulfide bonds in HIV-2 envelope glycoprotein, peptides expected to mimic all 11 disulfide-bonded domains were synthesized, oxidized or cysteine-alkylated; they were then purified and characterized. Rabbits were immunized with either linear cysteine-alkylated peptides (L1-L11) or cyclic oxidized peptides (C1-C11). All peptides except 7L elicited antibodies with titers between 10(3) and 5 x 10(6). Anti-peptide C (2, 3, 4, 7, 8, 9, 11) and anti-peptide L (2, 3, 8, 9, 11) antibodies recognized the native HIV-2 gp 125. Moreover, we found that cyclization of the peptides significantly increased the level of anti-peptide antibodies reacting with the intact antigen protein. Deglycosylation increased the level of protein reactivity of anti-peptide antibodies and rendered the epitopes in peptides 5, 6, 10 accessible, which were masked in the native protein. Peptide 1 induced antibodies reacting only with the denatured reduced gp125 HIV-2. In addition, while anti-peptide L antibodies reacted better with L peptide (called "linear" structural specificity), anti-peptide C antibodies reacted similarly with L and C peptides (called "broad" structural specificity). Interestingly, the "broad" structural specificity of antibodies correlated with reactivity against native gp125. Although none of these anti-peptide antisera displayed neutralizing activity against HIV-2ROD, these results support the hypothesis that the structural restriction of peptides have a major influence upon the generation of more specific antibodies for recognizing the intact protein.