Clonal deletion and functional inactivation of self-reactive cells have been invoked as mechanisms underlying intrathymic development of T-cell tolerance. The relative importance of these mechanisms in the development of tolerance of more mature, peripheral T cells either to self or to exogenous antigens is unclear, although recent data relate the development of T-cell tolerance in the periphery to clonal anergy. We have now investigated the induction of extrathymic tolerance using BALB/c mice that were made tolerant to Staphylococcus aureus enterotoxin B, a superantigen which specifically interacts in such mice with T cells bearing V beta 8 antigen receptors. Both euthymic and athymic mice made tolerant to S. aureus enterotoxin B had a markedly reduced number of V beta 8.1,2+ CD4+ peripheral T cells. This reduction was accompanied by genomic DNA fragmentation that is associated with cell death. These results indicate that a deletional mechanism can contribute to the induction of T-cell tolerance in peripheral lymphoid cells.