One tuberculosis vaccine candidate that has shown a promising degree of protective efficacy in guinea pigs is recombinant BCG Tokyo (Ag85A)(rBCG-Ag85A[Tokyo]). As a next step, cynomolgus monkeys were utilized because they are susceptible to Mycobacterium tuberculosis and develop a continuous course of infection that resembles that in humans both clinically and pathologically. The recombinant BCG vaccine was administered once intradermally in the back skin to three groups of cynomolgus monkeys, and its protective efficacy was compared for 4 months with that of its parental BCG Tokyo strain. Vaccination of the monkeys with the rBCG-Ag85A[Tokyo] resulted in a reduction of tubercle bacilli CFU (p<0.01) and lung pathology in animals challenged intratracheally with 3000 CFU H37Rv M. tuberculosis. Vaccination prevented an increase in the old tuberculin test after challenge with M. tuberculosis and reaction of M. tuberculosis-derived antigen. Thus, it was shown in monkeys that rBCG-Ag85A[Tokyo] induced higher protective efficacy than BCG Tokyo. This warrants further clinical evaluation.