We recently identified IgE-binding epitopes of bovine serum albumin (BSA), the major beef allergen, and found that the major epitope structure contains an EXXV motif. Here we report hypoallergenic analogue peptides of BSA that preserve T cell reactivity to retain the capacity to induce immunotolerance. Substitution of the glutamic acid in the EXXV sequence to aspartic acid leads to a remarkable reduction of IgE-binding ability and induces potent T cell proliferation. Moreover, in response to the substitutions, the production of INF-gamma is markedly increased compared with that elicited by the native peptides. Therefore, these analogue peptides represent potential candidates for specific immunotherapy.