Mucosal immunization can induce immune responses different from those induced by systemic immunization. In this study, murine immune responses were analysed after immunization through gastric and subcutaneous routes with Mycobacterium bovis BCG. The number of circulating cells secreting antigen-specific immunoglobulins (Ig), and the number of gamma interferon (IFN-γ) secreting cells in spleen-cell cultures after in vitro stimulation with mycobacterial antigens were analysed at the single cell level by the ELISPOT method. Levels of antigen-specific antibodies in sera were determined by ELISA. In the subcutaneously immunized mice the authors found approximately 100 times more splenic cells secreting antigen-specific IgG than in gastrically immunized mice or control mice. Their levels of antigen-specific IgG in sera were 66–6700 times higher than in sera from gastrically immunized mice or control mice. In contrast, the numbers of IFN-γ producing cells in spleen-cell cultures after in vitro activation with BCG were equally high in the immunized groups of mice, and for both groups higher than in non-immunized controls. Furthermore, IFN-γ producing cells could be demonstrated in gastrically immunized animals even without in vitro activation. The results demonstrate that gastric immunization with the BCG vaccine can induce a systemic T cell-mediated immune response against mycobacterial antigens.