EDIN-B (epidermal cell differentiation inhibitor-B; also termed C3Stau) is an exotoxin of Staphylococcus aureus which ADP-ribosylates and inactivates Rho GTP binding proteins. The EDIN-B gene (edin-B) and the gene for exfoliative toxin D (etd) make up the central part of a recently described pathogenicity island. Here we evaluated the prevalence and genetic organization of the edin-B/etd pathogenicity island in invasive S. aureus isolates, and characterized edin-B transcription and EDIN-B production using artificial constructs transduced in S. aureus strains RN6390 and Newman. We found that eight out of 121 (7 %) S. aureus blood culture isolates harbour edin-B, which is organized in three novel variants of the original edin-B/etd pathogenicity island. In the serum of patients infected with edin-B-positive S. aureus, significant titres of anti-EDIN-B antibodies could be detected. Regulation of edin-B transcription depended on the sarA but not on the agr regulatory system. Furthermore, retrieval of EDIN-B protein secreted by S. aureus RN6390 required the presence of alpha2-macroglobulin to inhibit the activity of extracellular proteases. These data suggest that the EDIN-B toxin is produced during human infection, is part of a highly variable pathogenicity island and can be controlled by the sarA gene regulon and secreted bacterial proteases.