A comparative study of the radioiodinated promastigote cell-surface antigens of Leishmania mexicana and L. major was carried out under reduced and nonreduced conditions by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by autoradiography. Under reduced conditions, the cell surface of L. mexicana promastigotes showed three iodinated polypeptides with molecular weights of 65,000, 50,000 and 27,000 daltons, whereas L. major promastigotes displayed a single polypeptide of 63,000 daltons. Under nonreduced conditions, the radioiodinated cell-surface component of L. major shifted to a mol.wt. of 51,000 daltons, whereas only one of the three components of L. mexicana (mol.wt., 65,000 daltons) underwent a large shift (to 59,000 daltons). The different immunochemical nature of the L. mexicana cell-surface antigens was demonstrated by using different anti-Leishmania sera. The rabbit anti-promastigote serum immunoprecipitated mainly the 50,000- and 27,000-dalton L. mexicana cell-surface polypeptides, whereas the rabbit anti-amastigote serum as well as a serum from a patient with cutaneous leishmaniasis immunoprecipitated almost exclusively the 65,000-dalton polypeptide. Immunoblot studies using a rabbit antibody against the L. major deglycosylated major surface antigen gp63 confirmed the differences in nature of the 65,000- and 50,000-dalton cell-surface antigens of L. mexicana. The results obtained are discussed in the light of the differences in antigenic cell-surface expression among Leishmania isolates and their consequences in the development of a differential diagnosis of leishmaniasis.