We have shown in a previous study that immunization of a rhesus monkey (Macaca mulatta) with inactivated liver stages of the simian malaria parasite Plasmodium cynomolgi (B strain) produced high antibody titers against sporozoites, liver stages, and blood stages of P. cynomolgi. In the present study, we demonstrate that these anti-P. cynomolgi immune sera recognized P. vivax (Salvador I) antigens. In an indirect immunofluorescence assay, both postimmunization and postchallenge sera reacted with antigens of sporozoite, liver-, and blood-stage parasites. In Western blot analysis, postimmunization sera recognized four bands of 97, 93, 70, and 65 kD in sporozoite antigens; postchallenge sera further recognized three doublet (set of two) bands of 86-90, 73-78, and 44-52 kD. When blood-stage extracts were used as antigens, five bands of 118, 105, 76, 68, and 47 kD reacted with postimmunization sera; two doublet bands of 81-87 and 49-52 kD further reacted with postchallenge sera. None of these sera reacted with asexual blood-stage extracts of P. falciparum and P. malariae, or with a recombinant circumsporozoite protein of P. vivax.