Extracellular vesicles (EVs) are particles formed by a lipid bilayer, released by various types of cells, plants and pathogens, with their main function being intercellular communication. EVs transport proteins, nucleic acids, and lipids and are potential candidates for clinical applications, therapies, and vaccines. EVs have been extensively studied as biomarkers for disease diagnosis and prognosis because they are found in most biological fluids and contain markers from the cell that released the particle. In the therapeutic field, modified EVs are used as drug delivery systems, offering advantages such as biocompatibility, immunogenicity, and the ability to cross biological barriers. Additionally, vaccines using EVs are being developed due to their immunomodulatory properties, which can induce protective immune responses against infectious diseases and cancer. Despite these advances, there are significant challenges, such as large-scale production, standardization, and approval by regulatory agencies. In this chapter, we describe the use of EVs as biomarkers, therapies, and vaccines against pathogens and cancer, as well as the challenges that scientists encounter in their application. Research on EVs is constantly advancing, and their use represents a revolution in precision medicine. Innovation in this field has the potential to transform diagnostic and therapeutic approaches to various diseases.
Extracellular vesicles (EVs) are emerging as key players in the pathogenesis of malaria and toxoplasmosis, two significant infectious diseases caused by Apicomplexa parasites. This chapter investigates the multifaceted roles of EVs in the progression of these diseases, emphasizing their involvement in immune modulation, hostparasite interactions, and the regulation of disease severity. In malaria, EVs derived from infected red blood cells, platelets, and endothelial cells contribute to disease symptoms, immune response modulation, and parasite survival and have potential as biomarkers and tools for vaccine development. Similarly, in toxoplasmosis, EVs influence the modulation of immune responses and disease progression, presenting distinct profiles depending on the Toxoplasma gondii strain. Notably, EVs from both parasites contain immunogenic proteins that can be used in vaccine development, with promising results in preclinical studies. The role of EVs in these parasitic infections highlights their potential as therapeutic targets and diagnostic tools, providing new opportunities for the prevention and treatment of malaria and toxoplasmosis.
Viruses are subcellular structures that depend on the host cell to replicate, sharing several characteristics with extracellular vesicles (EVs). EVs act in intercellular communication and the regulation of the immune system and can be exploited by viruses as vehicles for transport and dissemination between cells and organs. Although they may favor viral infection, EVs also stand out as potential biomarkers for the diagnosis of viral diseases, as they reflect the physiological and pathological state of the originating cells. These particles can contain viral RNA and specific proteins, allowing for the distinction between different types of infection. Moreover, EVs have great therapeutic potential and are being studied as nanotherapeutic tools due to their low immunogenicity, ability to cross cellular barriers, and ease of modification to allow delivery around the body and tissues. This chapter addresses the interactions between EVs and viruses, as well as the advancements in the use of these structures in diagnosis and the development of new therapeutic strategies. Understanding these mechanisms can significantly contribute to the control of viral infections and the creation of innovative therapies to treat emerging diseases.
Extracellular vesicles (EVs) are nano-sized, membrane-surrounded vesicles released by cells under both physiological and pathological conditions. Due to their small size and heterogeneity, comprehensive characterization of EVs remains technically challenging. Among the various analytical tools developed, flow cytometry stands out as a highly versatile and scalable platform, offering high-throughput analysis, multiparametric phenotyping, and quantitative detection. However, conventional flow cytometers are typically designed for cell-sized particles (0.5-40 µm) and require specific optimizations to reliably detect and analyze EVs, which are significantly smaller and result in weaker signals. These optimizations include instrument settings, sample handling and labelling strategies as well as acquisition protocols. Robust calibration and the use of appropriate controls are essential to ensure data accuracy and reproducibility across platforms. In this chapter, we outline the principles, technical considerations, and advantages of applying flow cytometry and imaging flow cytometry to EV research. We also highlight representative applications in both scientific and clinical contexts and discuss future directions for the field.
Extracellular vesicles (EVs) are membrane-bound nanostructures secreted by various cell types under physiological and pathological conditions. These vesicles carry a diverse cargo of biologically active molecules, including proteins, lipids, nucleic acids, and metabolites. The molecular and structural heterogeneity of EVs presents challenges in fundamental biology, biomarker development, and therapeutic applications. Fungal EVs have gained attention for their roles in pathogenesis, immune modulation, and potential targets for therapies and vaccines. EVs have numerous roles in intercellular communication, facilitated by the transfer of cargo to recipient cells or the interaction of EV surface proteins with cellular receptors. However, the question of how they traverse the cell wall remains a mystery. Fungal EVs can modulate the cell wall through enzymes, contributing to the transition of EVs by the fungal cell wall. As research progresses and technological barriers are overcome, EVs are emerging as valuable targets and promising tools in precision medicine. With continuous improvements in EV isolation, characterization, and manipulation, the next decade is likely to bring significant breakthroughs that will have a profound impact on both basic science and clinical practice.
