Herpes simplex virus 2 (HSV-2) is the main pathogen of genital herpes (GH).The currently available body of evidence suggests that only widespread use of an effective vaccine can control the spread of HSV-2.We developed a recombinant replication-defective adenovirus capable of expressing glycoprotein D2(rAd-gD2), and was identified by PCR and Western blot in gene and protein level, respectively. The mice in test groups were immunized intramuscularly (i.m.) with rAd-gD2 in 0and 4 week,while control mice received PBS and AdV. We detected gD2 IgG antibody and neutralizing antibody in order to evaluate of humoral immune response, detected Th1 (IL-2 and IFN-γ)/Th2 (IL-4, IL-5 and IL-10) cytokines in order to evaluate of cellular immune response. The results showed that immunization with rAd-gD2 could elicit strong gD2IgG(P<0.01)and neutralizing antibody levels(P<0.05),which were significantly higher than that with PBS and AdV groups IL-2IFN-γIL-4IL-5 and IL-10 detected in rAd-gD2 group were higher than control groups (P<0.01), Th1/Th2 cytokines were important standard of cellular/humoral immune response, respectively. The results demonstrated that rAd-gD2 could stimulate not only humoral immune response but also cellular immune response, rAd-gD2 may be good candidate for the control of HSV-2infection.