Extracellular cytolysis by activated macrophages: studies with macrophages on permeable membranes.

Mouse peritoneal macrophages were allowed to adhere to discs cut from permeable membranes, then activated by incubation in lymphokine-rich supernates from ConA-stimulated spleen cells. Such filter-borne cultures of activated macrophages (AM) were cytotoxic for various target cells (Tc). The kinetics of the cytotoxic process could be monitored by removal of the filter-bound AM after increasing times of contact with Tc. Using 3 assay procedures to assess macrophage cytotoxicity, i.e. chromium-51 release, thymidine incorporation, and cloning inhibition, most of the damage to Tc was found to occur within 30 min to 2 h of interaction between the two cell types. The kinetics of the cytolytic effect were similar, whether Tc were in direct contact with AM or separated by the filter; thus cytotoxicity appeared to be mediated by a highly diffusible compound. Supernates of AM incubated with Tc for 1 to 4 h, but not of AM incubated alone, were toxic for Tc, suggesting that Tc provide a signal to AM, in the absence of which toxic intermediates fail to be released. Addition of catalase or peroxidase considerably reduced Tc destruction by AM, indicating that oxygen metabolites might play a role as mediators of AM cytotoxicity in the present experimental model.