Recent studies have demonstrated that anti-DNA antibodies share important genetical features with antibodies to exogenous antigens, suggesting that anti-DNA antibody responses may be (auto-) antigen driven. We have earlier defined three out of five rabbits as anti-dsDNA antibody responders based on reactivity with calf thymus (CT) dsDNA after inoculation with the human dsDNA virus BK. In the present study we demonstrate that all five animals that received BK virus inoculations produced antibodies to BK virus dsDNA. These antibodies did not cross-react with CT dsDNA, as shown by inhibition experiments. The anti-BK dsDNA antibodies persisted over time, in contrast to the anti-CT dsDNA antibodies that decreased shortly after a peak following the first boost of BK virus. While the anti-CT dsDNA antibodies decreased, the anti-BK dsDNA antibodies remained elevated, thus supporting the results of the inhibition experiments which showed that two independent antibody populations are produced after BK virus inoculations. In the three animals producing anti-mammalian dsDNA antibodies, antibodies recognizing CT dsDNA reappeared after intravenous administration of a complex of CT dsDNA and methylated bovine serum albumin (MBSA) without adjuvant. The latter anti-CT dsDNA antibodies did not cross-react with BK dsDNA. In contrast to earlier studies we conclude that mammalian dsDNA may be immunogenic, and that discrete molecular differences in DNA antigens from different sources may induce anti-dsDNA antibodies specific for dsDNA molecules of different origin.