Neutrophils are rapidly recruited to the site of Leishmania infection and play an active role in capturing and killing parasites. They are the main source of leukotriene B4 (LTB4), a potent pro-inflammatory lipid mediator. However, the role of LTB4 on neutrophil infection by Leishmania amazonensis is not clear. In this study, we show that L. amazonensis or its lipophosphoglycan (LPG) can induce neutrophil activation, degranulation and LTB4 production. Using pharmacological inhibitors of leukotrienes synthesis, our findings reveal an LTB4-driven autocrine/paracrine regulatory effect. In particular, neutrophil-derived LTB4 controls L. amazonensis killing, degranulation and reactive oxygen species (ROS) production. In addition, L. amazonensis infection induces an early increase in Toll-like receptor (TLR)-2 expression, which facilitates parasite internalization. NFkB pathway activation represents a required upstream event for L. amazonensis-induced LTB4 synthesis. These leishmanicidal mechanisms mediated by neutrophil-derived LTB4 act through activation of its receptor, BLT1.