Peptides presenting predicted antigenic sites of CBV3 capsid proteins and peptide sequences from conserved regions of the nonstructural proteins were synthesized, and rabbit antipeptide sera were tested for their immunoreactivity. Peptides derived from different capsid regions were able to induce production of neutralizing antibodies in rabbits. As measured by EIA, all peptides representing four different proposed antigenic sites were immunogenic, inducing an antibody response against the homologous peptide and purified CBV3 as measured by EIA. Immunization with inactivated CBV3 induced a secondary response especially in rabbits primed with peptides representing polypeptide VP2. Antisera against the nonstructural protein sequences were highly cross-reactive with other enteroviruses, while the capsid peptide antisera were mainly type-specific when tested by immunoblotting against a panel of enteroviruses. Four of the capsid region peptides also exhibited distinct T-cell reactivity in a mouse T-cell proliferation assay.