title: Differential Interaction of the Staphylococcal Toxins Panton-Valentine Leukocidin and γ-Hemolysin CB with Human C5a Receptors.
abstract: Staphylococcus aureus is well adapted to the human host. Evasion of the host phagocyte response is critical for successful infection. The staphylococcal bicomponent pore-forming toxins Panton-Valentine leukocidin LukSF-PV (PVL) and γ-hemolysin CB (HlgCB) target human phagocytes through interaction with the complement receptors C5aR1 and C5aR2. Currently, the apparent redundancy of both toxins cannot be adequately addressed in experimental models of infection because mice are resistant to PVL and HlgCB. The molecular basis for species specificity of the two toxins in animal models is not completely understood. We show that PVL and HlgCB feature distinct activity toward neutrophils of different mammalian species, where activity of PVL is found to be restricted to fewer species than that of HlgCB. Overexpression of various mammalian C5a receptors in HEK cells confirms that cytotoxicity toward neutrophils is driven by species-specific interactions of the toxins with C5aR1. By taking advantage of the species-specific engagement of the toxins with their receptors, we demonstrate that PVL and HlgCB differentially interact with human C5aR1 and C5aR2. In addition, binding studies illustrate that different parts of the receptor are involved in the initial binding of the toxin and the subsequent formation of lytic pores. These findings allow a better understanding of the molecular mechanism of pore formation. Finally, we show that the toxicity of PVL, but not of HlgCB, is neutralized by various C5aR1 antagonists. This study offers directions for the development of improved preclinical models for infection, as well as for the design of drugs antagonizing leukocidin toxicity.
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands; Centre International de Recherche en Infectiologie, Université Lyon 1 and Ecole Normale Supérieure de Lyon, 69007 Lyon, France; Inserm, Unité 1111, 69007 Lyon, France;
authors.forename: András N
authors.initials: AN
authors.lastname: Spaan
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands;
authors.forename: Ariën
authors.initials: A
authors.lastname: Schiepers
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands;
authors.forename: Carla J C
authors.initials: CJ
authors.lastname: de Haas
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands;
authors.forename: Davy D J J
authors.initials: DD
authors.lastname: van Hooijdonk
authors.affiliation: Centre International de Recherche en Infectiologie, Université Lyon 1 and Ecole Normale Supérieure de Lyon, 69007 Lyon, France; Inserm, Unité 1111, 69007 Lyon, France; Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5308, 69007 Lyon, France;
authors.forename: Cédric
authors.initials: C
authors.lastname: Badiou
authors.affiliation: Cynbiose, 69280 Marcy l'Etoile, France;
authors.forename: Hugues
authors.initials: H
authors.lastname: Contamin
authors.affiliation: Centre International de Recherche en Infectiologie, Université Lyon 1 and Ecole Normale Supérieure de Lyon, 69007 Lyon, France; Inserm, Unité 1111, 69007 Lyon, France; Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5308, 69007 Lyon, France; Centre National de Référence des Staphylocoques, Hospices Civils de Lyon, 69007 Lyon, France;
authors.forename: François
authors.initials: F
authors.lastname: Vandenesch
authors.affiliation: Centre International de Recherche en Infectiologie, Université Lyon 1 and Ecole Normale Supérieure de Lyon, 69007 Lyon, France; Inserm, Unité 1111, 69007 Lyon, France; Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5308, 69007 Lyon, France; Centre National de Référence des Staphylocoques, Hospices Civils de Lyon, 69007 Lyon, France;
authors.forename: Gérard
authors.initials: G
authors.lastname: Lina
authors.affiliation: Ina Sue Perlmutter Laboratory, Division of Pulmonary Medicine, Department of Pediatrics, Boston Children's Hospital, Department of Medicine, Harvard Medical School, Boston, MA 02115; and Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA 02215.
authors.forename: Norma P
authors.initials: NP
authors.lastname: Gerard
authors.affiliation: Ina Sue Perlmutter Laboratory, Division of Pulmonary Medicine, Department of Pediatrics, Boston Children's Hospital, Department of Medicine, Harvard Medical School, Boston, MA 02115; and.
authors.forename: Craig
authors.initials: C
authors.lastname: Gerard
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands;
authors.forename: Kok P M
authors.initials: KP
authors.lastname: van Kessel
authors.affiliation: Centre International de Recherche en Infectiologie, Université Lyon 1 and Ecole Normale Supérieure de Lyon, 69007 Lyon, France; Inserm, Unité 1111, 69007 Lyon, France; Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5308, 69007 Lyon, France;
authors.forename: Thomas
authors.initials: T
authors.lastname: Henry
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands; j.vanstrijp@umcutrecht.nl.
authors.forename: Jos A G
authors.initials: JA
authors.lastname: van Strijp
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000276:immunology*; D001427:Bacterial Toxins / Q000276:immunology*; D002417:Cattle; D002460:Cell Line; D005098:Exotoxins / Q000276:immunology*; D057809:HEK293 Cells; D006460:Hemolysin Proteins / Q000276:immunology*; D006801:Humans; D057131:Immune Evasion / Q000276:immunology; D007956:Leukocidins / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000276:immunology; D010586:Phagocytes / Q000276:immunology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D044087:Receptor, Anaphylatoxin C5a / Q000037:antagonists & inhibitors / Q000276:immunology*; D019707:Receptors, Chemokine / Q000037:antagonists & inhibitors / Q000276:immunology*; D013203:Staphylococcal Infections / Q000276:immunology; D013211:Staphylococcus aureus / Q000472:pathogenicity
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; C488033:C5AR1 protein, human; C417003:C5aR2 protein, human; D005098:Exotoxins; D006460:Hemolysin Proteins; D007956:Leukocidins; C078284:Panton-Valentine leukocidin; D044087:Receptor, Anaphylatoxin C5a; D019707:Receptors, Chemokine; C037956:gamma-hemolysin, Staphylococcus aureus