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The lack of understanding of the requirements for effective and efficient intracellular Leishmania amazonensis killing by activated macrophages has been a barrier to vaccine development against this parasite. Here we report using inhibitors of superoxide and inducible nitric oxide synthase that the presence of both superoxide and nitric oxide is necessary for killing of L. amazonensis amastigotes within LPS/IFN-y activated bone marrow-derived macrophages generated from C3H mice. Nitric oxide alone was sufficient to control L. major infection, as addition of a nitric oxide donor was sufficient to control L. major but not L. amazonensis infection. Addition of an extracellular signal-regulated kinase (ERK) inhibitor to L. amazonensis-infected macrophages increased the ability of these activated macrophages to kill L. amazonensis amastigotes. This enhanced macrophage killing through addition of ERK inhibitor was abrogated by inhibition of superoxide or iNOS, indicating that ERK activation may modulate effective macrophage killing lending to the ability of Leishmania amazonensis to resist elimination within activated macrophages.

Leishmania amazonensis and macrophage interactions: immune factors necessary to kill the parasite