This study examined the immune responses induced by cruzipain, a well-characterized T. cruzi antigen, to determine whether it is a relevant immunogen among the parasite acidic antigens (FIII), for which some biological properties have been studied previously. Humoral and cellular immune responses were investigated in BALB/C mice after immunization with cruzipain or FIII. Skin tests revealed immediate type-hypersensitivity (ITH) and delayed-type hypersensitivity (DTH) reactions to cruzipain in both groups of immunized mice. IgG1 and IgE isotypes against cruzipain were detected by ELISA in both groups and immunoblot studies showed that these antibodies recognized a major protein band of 50 kDa, cruzipain. The antigen-specific proliferative responses of spleen lymphocytes from both groups of immunized mice were also increased. Immunization with cruzipain of FIII antigen significantly enhanced the percentage survival of mice challenged with 10(3) trypomastigotes. The results revealed high cross-reactivity between cruzipain and FIII, suggesting the cruzipain is a relevant immunogen among the parasite acidic antigens.