The potential of staphylococcal enterotoxin A (SEA), B (SEE), C (SEC) and toxic shock syndrome toxin-l (TSST-1) to act as superantigens by inducing polyclonal T-cell mitogenesis and cytokine production was tested on bovine peripheral blood mononuclear cells (PBMC). These four toxins were capable of inducing strong proliferative response of PBMC from carves over a broad dosage range (1 pg/ml to 1 mu g/ml) in vitro. The toxin-activated blast cells consisted of both CD4(+) T-cells and CD8(+) T-cells, but the T-cell proliferation depended upon the presence of monocytes. Treatment of monocytes with monoclonal antibody to major histocompatibility complex class II antigens substantially inhibited the toxin-induced T-cell proliferative response, but paraformaldehyde-fixation did not abrogate the accessory function. SEA, SEE, SEC and TSST-1, all induced the in vitro release of interleukin-2, interferon gamma and tumor necrosis factor cu in a dose dependent manner. The results indicate that SEA, SEE, SEC and TSST-1 are capable of acting as superantigens by stimulating bovine T-cells as shown in the human and murine systems. The possible implications of these toxins in the immunopathogenesis of bovine mastitis caused by the infection with Staphylococcus aureus are discussed.