Generation of cytotoxic T lymphocytes (CTL) in vitro and tumor-rejection responses by sensitization of semi-syngeneic mice with tumor-antigen-reconstituted liposomes were investigated. Liposomes were prepared from a crude butanol extract (CBE) of BALBRVD leukemia cells and egg phosphatidylcholine (PC): 1,2-dimyristoylamido-1,2-deoxyphosphatidylcholine (DDPC) (3:2) or dimyristoylphosphatidylcholine (DMPC):DDPC (1:4). Inter-membrane protein transfer (IMPT) liposomes were prepared by incubating BALBRVD cells with DMPC:DDPC (1:4) liposomes. Sensitization of male CB6F1 mice with CBE or IMPT liposomes induced a level of cytotoxicity similar to that on sensitization with mitomycin-C(MMC)-treated BALBRVD against BALBRVD target cells after in vitro sensitization with the tumor cells. Sensitization with CBE alone resulted in only marginal cytotoxicity. The cytotoxic effector cells induced by either mode of sensitization were CD8+ T-cells whose recognition was Kd-restricted. No difference in specificity was observed with the different modes of sensitization. Two in vivo immunizations with CBE or with CBE liposomes at a dose of 25 micrograms of protein (equivalent to 2.5 x 10(7) cells) cause moderate inhibition of BALBRVD tumor growth in male CB6F1 mice and immunization with IMPT liposomes at a dose of 1 microgram of protein result in efficient protection.