Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis

Here, we present a protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis. We describe steps for Leishmania infection, harvesting infected tissues, and performing liquid chromatography-mass spectrometry (LC-MS) to identify enriched metabolic pathways and altered metabolites. We then detail procedures for verifying the metabolite role in immune response via PCR and ELISA. This protocol focuses on cutaneous leishmaniasis models, but the analyses here are applicable to other infection models. For complete details on the use and execution of this protocol, please refer to Volpedo et al.(1) and Oljuskin et al.(2).