It was recently shown that immunization of hamsters with DNA plasmids coding LJM19, a sand fly salivary protein, partially protected against a challenge with Leishmania chagasi, whereas immunization with KMP11 DNA plasmid, a Leishmania antigen, induced protection against L. donovani infection. In the present study, we evaluated the protective effect of immunization with both LJM19 and KMP11 DNA plasmid together. Concerning the protection against an infection by L. chagasi, immunization with DNA plasmids coding LJM19 or KMP11, as well as with both plasmids combined, induced IFN-γ production in draining lymph nodes at 7, 14 and 21 days post-immunization. Immunized hamsters challenged with L. chagasi plus Salivary Gland Sonicate (SGS) from Lutzomyia longipalpis showed an enhancement of IFN-γ/IL-10 and IFN-γ/TGF-β in draining lymph nodes after 7 and 14 days of infection. Two and five months after challenge, immunized animals showed reduced parasite load in the liver and spleen, as well as increased IFN-γ/IL-10 and IFN-γ/TGF-β ratios in the spleen. Furthermore, immunized animals remained with a normal hematological profile even five months after the challenge, whereas L. chagasi in unimmunized hamsters lead to a significant anemia. The protection observed with LJM19 or KMP11 DNA plasmids used alone was very similar to the protection obtained by the combination of both plasmids.Graphical abstractImmunization of hamsters with LJM19 and/orKMP11 lead a decrease in the parasite load in the spleen and liver in hamsters challenged with L. chagasi plus Lutzomyia longipalpis and maintenance of hematological parameters.Highlights► Immunization with LJM-19 and/or KMP11 induces protection in hamsters (L. chagasi). ► The immunization induces high IFN-γ/IL-10 and IFN-γ/TGF-β in draining lymph nodes. ► Two and 5 months after challenge, immunized animals showed lower parasite load in liver and spleen. ► Immunized animals remained with a normal hematological profile.