Spleen is one of the largest lymphoid organs in the body; it harbors immune cells including antigen presenting cells, B and T lymphocytes. It has an important role in humoral and cellular immune responses. Herein we investigated the role of spleen in the immune response to experimental Leishmania major infection. It is known that C57BL/6 mice are resistant to L. major infection whereas BALB/c mice are susceptible. Although splenectomy was associated with reduced serum levels of IFN-gamma, absence of the spleen did not change the profile of L. major infection in the resistant C57BL/6 and BALB/c susceptible mice. Both strains of mice maintained the same profile of cytokine production in regional lymph nodes after splenectomy and responded in the same way against the infection. Only splenectomized BALB/c mice had a reduction in IL-4 and IL-10 production by lymph node cells early in infection. Our data suggest that, in localized infections, regional lymph nodes may replace efficiently the immunological role of spleen in the cellular and humoral immune responses.