The hypothesis that natural killer (NK) cells represent an important form of cell recognition and cytotoxicity leads to the prediction that NK-like activity should be preserved throughout phylogenetic development. This was tested in the invertebrate Planorbarius corneus. Two types of cells can be identified and separated from the hemolymph of this mollusc, i.e. glass-adherent macrophage-like spreading hemocytes (SH) and nonadherent round hemocytes (RH). Only RH are able to lyse the K-562 human target cell line in a short-term NK cytotoxicity test. This NK-like activity, severely reduced after 18 h incubation at 24 degrees C, is preserved by human recombinant interleukin 2. A further analysis of P. corneus hemocytes has been performed by using several mouse anti-human monoclonal antibodies and cytofluorimetric analysis. Unexpectedly, both SH and RH react with several monoclonal antibodies, including those directed against epitopes typical of mammalian NK cells and cell adhesion molecules. On the whole, these data support the hypothesis that a primitive NK-like activity appeared early in evolution and is not shared by phagocytic cells.