Sulfatide induces leukocyte activation, which is thought to be mediated via L-selectin. Here we demonstrate that lymphocytes express a receptor for sulfatide distinct from L-selectin and that this receptor is involved in sulfatide-induced cell activation. While cell surface L-selectin expression was abolished by phorbol 12-myristate 13-acetate (PMA), lymphocytes retained the ability to bind sulfatide in liquid phase as well as in immobilized solid phase. The novel sulfatide receptor obtained from PMA-treated lymphocytes showed a molecular size of 65 kDa. Stimulation through this receptor triggered cytosolic free Ca 2+ elevation and intercellular aggregation.