Leishmaniasis is caused by intracellular parasites of Leishmania species, which are transmitted by the bite of the sandfly. Recovery and protection against the infection depends on the induction of a strong Th1 type of immune response. Vaccination of mice with the opioid antagonist naloxone can promote the activation of the Th1 responses. We studied the efficacy of the mixture of naloxone and alum, as an adjuvant, to enhance immune responses and induce protection against Leishmania major infection in BALB/c as a susceptible mouse model. BALB/ c mice were immunized with Ag-naloxone-alum, Agalum, Ag-naloxone or PBS subcutaneously three times at 2-week intervals. The humoral and cellular specific immune responses were assessed 2 weeks after the last immunization and compared with the control mice. Our results indicated that the administration of alum-naloxone as an adjuvant increased the capability of L. major promastigote antigens to enhance lymphocyte proliferation, the levels of IFN-c, and the IFN-c/IL-5 ratio. The results of DTH showed that there were no significant differences in footpad swelling between the groups of immunized mice as compared with the non-vaccinated control group; however, no significant differences were observed in the survival rate among groups. It can be concluded that although immunization with the alum-naloxone mixture in combination with the autoclaved L. major promastigote antigens could enhance cellular immunity and shift the immune response to a Th1 pattern, it could not protect the mice against Leishmania major infection.