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Sand flies are bloodsucking insects transmitting parasites of genus Leishmania, the causative agents of diseases in humans and dogs. Experimental hosts repeatedly exposed to sand fly saliva can control Leishmania infection. Cell-mediated anti-saliva immune response is most likely responsible for this protective effect; however, there is no study so far concerning its antigenic specificity towards different sand fly vectors. In this study, splenocytes from BALB/c mice repeatedly exposed to the bites of Phlebotomus sergenti were challenged ex vivo with salivary gland homogenates from three different sand fly vectors -P. sergenti, P. papatasi, or P. arabicus. Mice bitten by P. sergenti had higher proliferative response to homologous antigen than splenocytes from naive mice. Splenocytes from P. sergenti bitten mice as well as anti-P. sergenti antibodies partially cross-reacted with P. papatasi saliva. In contrast, no cross-reactivity was found with P. arabicus saliva. Our data indicate that both arms of the immune system, cellular and humoral, react in a species-specific manner. Therefore, the presence of antibodies against salivary components of a certain species indicates the specificity of cell-mediated immune response as well. The data suggest that unique transmission-blocking vaccine would be required for each vector -Leishmania combination.