Exosomes are extracellular vesicles of 50–150 nm in diameter secreted by basically all cell types. They mediate micro-communication among cells, tissues, and organs under both healthy and disease conditions by virtue of their ability to deliver macromolecules to target cells. Research on exosomes is a rapidly growing field, however many aspects of their biogenesis and functions still await a complete clarification. In our review we summarize most recent findings regarding biogenesis, structure, and functions of exosomes. In addition, an overview regarding the role of exosomes in both infectious and non-infectious diseases is provided. Finally, the use of exosomes as biomarkers and delivery tools for therapeutic molecules is addressed. Considering the body of literature data, exosomes have to be considered key components of the intercellular communication in both health and disease.