The mechanisms of rejection of secondary tumours were studied in concomitant immunity using transplants of standard-size solid SL2 tumours in syngeneic DBA/2 mice. In such a system primary tumor implants are not rejected, in contrast to secondary tumour implants. The second tumour was mainly rejected 2-4 days after implantation. Both primary tumour and secondary tumour implants (which are rejected) contained hardly any lymphocyte infiltrate, whereas, 2-4 days after implantation they contained 40%-50% macrophages, which were cytotoxic in vitro. Transfer (s.c.) of these tumours to naive mice showed that cellular infiltrates in the secondary implants did not always cause tumour rejection. Serum collected on day 4 after implantation of the secondary tumour was cytotoxic to SL2 tumour cells in vitro, whereas serum from mice with only primary implants was not cytotoxic on day 4 after implantation. Preliminary characterization of this cytotoxic factor showed that it was heat-labile, as cytotoxicity disappeared after 30 min at 56 degrees C, the molecular mass of the factor was higher than 100 kDa, and it was not IgG. We hypothesize that secondary tumours in the DBA/2-SL2 concomitant immunity system are rejected mainly between 2 and 4 days after implantation as a result of the combined action of cytotoxic serum and the presence of 40%-50% cytotoxic macrophages. The primary tumour is not rejected at 2-4 days after implantation as there is no cytotoxic factor.