Ventricular assist devices (VADs) are an important form of therapy for end-stage congestive heart failure. However, infection of the VAD, which is often caused by Staphylococcus aureus, poses a major threat to survival. Using a novel in vitro binding assay with VAD membranes and a heterologous lactococcal system of expression, we identify 3 S. aureus proteins-clumping factor A (ClfA) and fibronectin binding proteins A and B (FnBPA and FnBPB) as the main factors involved in adherence to VAD polyurethane membranes. Adherence is greatly diminished by long implantation times, reflecting a change in topological features of the VAD membrane, and is primarily mediated by the FnBPA domains in the staphylococcal proteins. We also compare the adherence of S. aureus mutant strains and show that other staphylococcal components appear to be involved in adherence to VAD membranes. Finally, we demonstrate that ClfA, FnBPA, and FnBPB mediate bacterial infection of implanted murine intra-aortic polyurethane patches.The ventricular assist device (VAD) is an important form of therapy for patients with congestive heart failure. Originally introduced as a bridge to cardiac transplantation, it is increasingly also used for "destination therapy" [1]. A serious limitation to the long-term use of VADs has been the high incidence of device-related infections, which occur in 28%-48% of patients [2][3][4][5]. These infections, which are often caused by Staphylococcus aureus, pose a major threat to survival, because eradication of the infection usually requires removal of the device [2,6,7].Although the clinical features of VAD-related infections caused by S. aureus are well described, little is known about their pathogenetic processes [2,[5][6][7]. The microenvironment of the VAD surface is dynamic, with constant remodeling of the neointimal surface, beginning with platelet deposition and formation of a fibrin scaffold after which a variety of cell types attach and proliferate [8,9]. As a result of this process, the bacterialneointimal surface interactions change over time. Despite the increasing biological relevance of these interactions, the complex interplay of bacterial adhesins and the prosthetic neointimal surface has received limited attention. In particular, the contribution of proteins belonging to the family of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), which have been demonstrated to play a role in other S. aureus inDownloaded from https://academic.oup.