The intracellular location of some micro-organisms has been early recognised as a critical point to explain failure of antibiotic therapy to eradicate such pathogens from infected hosts. Most often parasites invade 'professional' phagocytic cells, including neutrophils, monocytes and macrophages, by resisting the intracellular bactericidal phagolysosomal pathway. Alternatively, they may invade 'non-professional' phagocytic cells (cells with fewer phagocytic and bactericidal abilities) such as endothelial cells, or even cells without lysosomes such as erythrocytes. The intracellular activity of an antibiotic depends on several factors including its ability to reach the eukaryotic cell membrane, its subcellular localisation as compared to that of the parasite, the possibility that the intracellular milieu may partially inactivate its activity, and the susceptibility of the intracellular form of the parasite. In vitro and animal models have been developed to investigate antibiotic activity against intracellular pathogens. However, it should be emphasised that only data obtained from patients give reliable information to define the optimum antibiotic regimen.