We evaluated the prime-boost immunization consisting of hepatitis C virus (HCV)-core expression plasmid (pCEP4-core) and replication-defective adenovirus expressing HCV-core (Adex1SR3ST) for core-specific CTL induction in mice. Compared to a single booster, double boosters after priming enhance CTL induction. The prime-double boosts immunization involving pCEP4-core priming followed by pCEP4-core and Adex1SR3ST boostings (pC/pC/aC) can induce core-specific CTLs as well as other combinations: pC/aC/aC; aC/pC/pC; aC/aC/aC, whereas pC/pC/pC does not induce CTLs. Furthermore, co-administration of interleukin-12 (IL-12) expression plasmid leads to the highly efficient CTL induction and clearance of HCV-core expressing vaccinia virus challenged. Thus, the prime-double boosts immunization together with IL-12 may be promising for HCV vaccine.