Chimeric dengue type 2/type 1 (DEN2/1) viruses, which contain the structural genes of the dengue-1 (16007) parental virus and the nonstructural genes of the DEN2-PDK53 virus, have been constructed. These DEN2/1 viruses induce high levels of DEN1 virus-specific neutralizing antibodies in mice. In this study, the DEN2/1 viruses induced DEN1 virus-specific neutralizing antibodies without the development of viremia in cynomolgus monkeys. Dengue virus-specific IgM antibodies were detected in the sera of the immunized animals as early as 3 days post-immunization. After challenge with the DEN1-16007 wild-type virus, only a low level of viremia was detected in chimeric DEN2/1 virus-immunized monkeys. A second challenge, with DEN2-16681 virus, was given while the levels of DEN2-specific neutralizing antibodies were very low: infectious Dengue 2 virus could not be detected in sera of the monkeys. A correlation between the level of neutralizing antibody and the incidence of viremia could not be found. In addition, there was no significant increase in the levels of interferon gamma and soluble interleukin 2 receptor in the sera of the challenged monkeys, which suggests a reduction in immunopathogenesis caused by T-cell activation. Our findings suggest that DEN2/1 viruses may used as a live-attenuated candidate vaccine because of their safety, broad immunogenicity, and lower immunopathogenicity.