Crayfish hemocytes displayed cytotoxic capacity towards all tested mammalian tumor and nontumor cell lines. The ratio required for the cytotoxic action of effector cells to target cells was at least 1:1. The lysis of the target cells required a minimum of 1 hr to become detected. After separation and isolation of the hemocyte populations of crayfish, the semigranular and granular cells retained their cytotoxic capacity. These cells contain the prophenoloxidase activating (proPO) system, a complement-like pathway, which in an activated form lyses semigranular cells in vitro, but failed to kill the tested target cells.