Leishmania parasites cause a spectrum of diseases that afflict the populations of 88 countries around the world and all attempts to control leishmaniasis have failed. It seems that preparing a vaccine may be the final solution. The aim of this study was to determine various Leishmania (L.) major antigens vaccine candidates and effects of the vaccine on delayed type hypersensitivity (DTH). Many different methods of vaccine preparation plus or without adjuvant were used. We prepared crude antigen combinations by five different methods using antigens from L. major parasites. Phase I was done in animals. The immunogenic effect was evaluated with the delayed type hypersensitivity (DTH) reaction with five different doses, including 100, 200, 300, 400, 500 microg/ml of total protein + BCG in Balb/c and conventional laboratory white mice (out breed). Our results showed that the cocktail antigen was highly specific. No injection of BCG solvent or saline treated controls showed significant results. Taken together, the effect of cellular immune response to the cocktail vaccine induced a significant effect against cutaneous leishmaniasis in the experimental model of vaccine with L. major.