Human cutaneous leishmaniasis (CL) is an important public health problem worldwide, being endemic in Latin-America. The clinical features are localized skin lesions that can resolve spontaneously or by antimonial treatment, but can become chronic, leading to severe tissue destruction and disfigurement. Host cells are mainly macrophages and dendritic cells and the resolution of the lesions depends on the magnitude of the innate and adaptive immune responses generated by the human host. The outcome of the disease is determined by the destruction of the parasite carried out by the host cells activated by T lymphocytes, direct action by cytotoxic cells and the cytokine network. Communication between cells is essential to maintain homeostasis and respond adequately to cell damage and invasion of pathogens. This communication can be mediated by direct cell-to-cell interaction, but it is often driven by hormones, cytokines and inflammatory mediators. EVs, like exosomes and microvesicles (MVs), are able to transfer regulatory molecules and genetic information from one cell to another. Since EVs are formed by invaginations of the plasma membrane or the endosomal system and released by most nucleated cells, and because their composition changes under different physiological and pathological conditions, they are defined as important regulators of cellular functions and pathogenesis mechanisms. Nano-Flow Cytometry is useful for identifying, phenotyping and counting MVs in plasma and cell culture supernatants. In addition, as the therapeutic approaches used worldwide are unable to achieve a completely sterile cure and there is no vaccine available, investigating the contributions of EVs in the pathological conditions of CL is essential to generate knowledge to define their potential roles in the immunopathogenesis and as biomarkers and therapeutic targets.