A vaccine against HIV-1 infection absolutely needs the ability to effectively elicit virus-specific immunity over a long term; nevertheless, there have been few studies indicating that the immunoinductivity of such a candidate vaccine has been researched for several years running. In a previous report, we demonstrated that recombinant BCG (rBCG) expressing the full-length gag gene of simian immunodeficiency virus (SIV) (rBCG-SIVgag) induced Gag-specific delayed-type hypersensitivity, T cell proliferation, gamma interferon (IFN gamma), and serum IgG responses in guinea pigs immunized intradermally (i.d.) with 0.1 mg for the 1-year period of study. Especially, the production of long-lasting Gag-specific serum IgG in the vaccinated animals perhaps reflects the persistent antigenic stimulation by rBCG-SIVgag. How long, we questioned, will such immune responses to Gag engendered by the rBCG-SIVgag vaccination persist without booster immunizations? To learn this, we examined Gag-specific IgG production in sera and Gag-specific IFN gamma mRNA expression in peripheral blood mononuclear cells (PBMC) in guinea pigs vaccinated with rBCG-SIVgag i.d. (0.1 mg) or orally (80 mg x 2) during a 3-year period. As a result, Gag-specific serum IgG was highly generated for 3 years at similar levels between the i.d. and the orally immunized guinea pigs (IgG2>IgG1). The enhancement of IFN gamma mRNA expression by in vitro restimulation with Gag antigen was also detected in PBMC from the two immunization groups throughout the 3-year observation period. In guinea pigs immunized i.d. with rBCG-SIVgag, a high level of Gag-specific IFN gamma response was observed at 1 year after vaccination, whereas the response has waned gradually. The current study indicates that i.d. and oral inoculations of rBCG-SIVgag elicit stable, strong, Gag-specific serum IgG production while exhibiting the different kinetics of Gag-specific IFN gamma responses between i.d. and oral vaccination routes. This suggests that the rBCG vector system expressing an appropriate size of foreign antigen gene should be suited for the induction of the antigen-specific humoral immunity concomitant with an IFN gamma response.