In contrast to L. major, the factors required for clearance of Leishmania amazonensis parasites from infected macrophages have been difficult to define. Multiple studies have made progress towards identifying the phenotypic differences in various cell types secondary to L. amazonensis infection as compared to L. major infection, but few have shown the cell types or factors required for parasite clearance. Based on studies which identified that mice previously infected with L. major and healed can mount a protective immune response against L. amazonensis, this study identifies cell types and factors from draining lymph node cells of L. major-infected mice that are necessary and sufficient to control infection in L. amazonensis-infected bone-marrow derived macrophages. Using a transwell system we show that soluble factors from CD4+T cells and B cells were required to kill intracellular parasites. One of these factors, L. major-specific immunoglobulin, may serve to trigger macrophage activation and promote parasite killing via superoxide production. Identification of these factors will provide more precise knowledge of host-cell signaling required to promote an effective immune response against L. amazonensis.