Cellular responses to several purified antigens of Mycobacterium bovis were examined in experimentally infected cattle over a period of 36 months, using in vitro cellular proliferation and interferon-gamma assays. These antigens (12, 19, 22a, b, 24, 25, 30, 32, 39, 65 and 70 kDa) included the majority of M. bovis protein antigens described to date and are highly homologous to those purified from M. tuberculosis. Cellular responses were examined at 3-month time intervals during the 36-month course of infection. All purified antigens induced cellular immune responses in the infected animals. The onset and magnitude of response to individual antigens varied among the animals. At any specific time during the period of infection one or more antigens appeared to be immunodominant but the immunodominance profile changed as the infection progressed. Humoral immune responses were low or absent in the first half of the infection period, but increased substantially for some of the antigens during the second half. Variation was observed among the different animals as to which antigens they recognized.