Staphylococcus aureus is the leading cause of infective endocarditis in the United States. Infective endocarditis (IE) is defined as an infection of the endocardium, typically involving the heart valves. The hallmark features of IE are vegetations. Vegetations are cauliflower-like, stratified biofilms of bacteria and host factors that develop on the valve leaflets of the heart. The mechanisms of how vegetations form are not well understood, and as a consequence the bacterial factors that are important for development of IE are not well defined. My studies focus on the role of a family of S. aureus exoproteins known as superantigens and their role in IE. Superantigens (SAgs) are a class of secreted virulence factors that have been extensively studied for their role in systemic diseases such as toxic shock syndrome (TSS), pneumonia, and food poisoning. The SAg protein family is comprised of 23 distinct members designated as staphylococcal enterotoxin (SE) or enterotoxin-like (SEl) and toxic shock syndrome toxin-1 (TSST-1). The term superantigen is derived from the ability of SAgs to interact with the immune system, resulting in a nearly 3000-fold increase in activation when compared to standard antigens. SAgs have a defined structure that is composed of 2 domains, a carboxy-terminal beta-grasp domain and amino-terminal oligosaccharide/oligonucleotide binding (OB) fold. Defined groups of SAgs are associated with S. aureus strains isolated from specific diseases, but few studies have been done to determine the role of SAgs in diseases outside of TSS and food poisoning. The enterotoxin gene cluster (egc) is a group of 6 SAgs (selo, selm, sei, selu, seln, and seg) assembled into an operon-like cluster that is present in the majority of S. aureus strains isolated from IE patients. My studies have determined that the egc is able to induce vegetations when expressed in avirulent S. aureus strains. This is the first time the egc has been directly associated with IE. I further characterized the capacity of the individual egc proteins to induce vegetations. Four (selo, selm, sei, and selu) of the 6 egc SAgs were able to induce vegetation formation. This is the first time the individual egc proteins have been characterized and directly associated with IE. I also demonstrated that the egc proteins may not be exclusively expressed as a single polycistronic transcript but that selu and seg contain promoter elements that may drive their individual expression. Lastly, I provide evidence that the egc SAgs may be regulated by MgrA, a global regulator of S. aureus associated with virulence factor expression.

Du fait de leurs propriétés immunostimulantes, les entérotoxines de Staphylococcus aureus (SEs) sont aussi considérées comme des outils thérapeutiques anticancéreux potentiels. Cependant, leurs implications dans de nombreuses pathologies humaines limitent leurs utilisations. Récemment, un opéron dénommé enterotoxin gene cluster (egc) codant pour cinq entérotoxines (SEG, SEI, SElM, SElN et SElO) supposées être de moindre virulence pour l’organisme, a été mis en évidence. En 2004, des patients atteints de carcinome broncho-pulmonaire ont été traités par l’administration d’un surnageant de culture d’une souche de S. aureus, contenant l’opéron egc. Ce traitement a permis d’allonger la durée de survie, et n’a eu aucun effet secondaire. Dans ce cadre, l’objectif de cette thèse a été d’étudier l’activité anti-tumorale des toxines de l’egc. Nos travaux ont mis en évidence l’activité tumoricide de ces toxines, induite par l’activation du système immunitaire. Cette toxicité est médiée par la sécrétion de nombreux médiateurs solubles comme le TNF-α et le NO. Nous avons confirmé le caractère pro-inflammatoire de type Th1 des toxines de l’egc. Nos travaux ont également montré qu’hormis SEI, les toxines de l’egc induisent des sécrétions de cytokines, chimiokines, protéases matricielles (MMPs) et facteurs de croissances nettement inférieures à celle induites par le reste des SEs. Ces résultats pourraient expliquer la faible toxicité associée aux toxines de l’egc. Enfin, nous avons montré que SElO possèdent une toxicité intrinsèque vis-à-vis des lignées tumorales. Cette étude plaide en faveur de l'intérêt des toxines de l’egc dans le développement de nouvelles approches en thérapie anti-tumorale / The use of classical superantigens (e.g. SEA, SEB and SEC) for treatment of cancer has resulted in a low response rates due to serious toxicity in humans. However, in a recent clinical study, remarkable results in treating lung cancer were obtained using superantigens encoded by the enterotoxin gene cluster (egc) without causing any significant toxicity. The current study was performed to investigate how egc superantigens (i.e. SEG, SEI, SElM, SElN and SElO) have tumoricidal activity with low toxicity. Indeed, we first demonstrated that tumoricidal activity of egc-SEs is mediated by immune cell activation, in particular, by secretion of soluble mediators such as nitric oxide and TNF-α. Thus, the proteomic analysis of the PBMC supernatants, showed that SEs-egc enhance the expression of pro-inflammatory cytokines, chimokines and many other biomarkers. Interestingly, levels were significantly higher in supernatants of SEA-stimulated PBMC than those with egc superantigens suggesting that staphylococcal superantigens differs in their inflammatory proprerties. Our results suggest that the relative lower pro-inflammatory activity of egc toxins may explain the low toxicity of these toxins observed during the clinical trial. Finally, we showed that SElO have a direct cytostatic activity against tumor cells. These findings suggest that egc-SEs seems to be good candidates for the development of new drugs in cancer therapy