We investigated whether the immunogenicity of antigens delivered by recombinant E1-deleted adenovirus (Ad) is impaired by the concomitant priming of specific immunity to protein antigens of the vector. A comparative evaluation of the immunogenicity of hepatitis B surface antigen (HBsAg or S) or ovalbumin (OVA) was carried out in mice injected with either the antigen-encoding Ad vector or a corresponding plasmid DNA vaccine. Recombinant Ad, but not the plasmid DNA vaccine, induced long lasting, specific CD8 T-cell immunity to immunodominant epitopes of the two antigens. In contrast, the HBsAg-encoding pCI/S DNA, but not the Ad/S vaccine, was shown to prime CD8 T-cell responses to subdominant HBsAg epitopes. Ad/S-primed CD8 T-cell responses to immunodominant epitopes of vector-encoded or capsid-delivered Ad proteins apparently suppressed CD8 T-cell priming to subdominant HBsAg epitopes. In B-cell-deficient mice, the established, Ad-specific T-cell immunity induced by vaccination with an irrelevant Ad vector impaired the priming of HBsAg-specific CD8 T-cell responses by Ad/S. It is clear, therefore, that a T-cell immunity specific for Ad proteins (either delivered with the Ad capsid or transcribed from the Ad genome) is efficiently primed by vaccination with Ad vectors, and can limit the immunogenicity (particularly of subdominant epitopes) of Ad vector-encoded transgenic antigens.