title: Contribution of the collagen adhesin Acm to pathogenesis of Enterococcus faecium in experimental endocarditis.
abstract: Enterococcus faecium is a multidrug-resistant opportunist causing difficult-to-treat nosocomial infections, including endocarditis, but there are no reports experimentally demonstrating E. faecium virulence determinants. Our previous studies showed that some clinical E. faecium isolates produce a cell wall-anchored collagen adhesin, Acm, and that an isogenic acm deletion mutant of the endocarditis-derived strain TX0082 lost collagen adherence. In this study, we show with a rat endocarditis model that TX0082 Deltaacm::cat is highly attenuated versus wild-type TX0082, both in established (72 h) vegetations (P < 0.0001) and for valve colonization 1 and 3 hours after infection (P <or= 0.0002), making Acm the first factor shown to be important for E. faecium pathogenesis. In contrast, no mortality differences were observed in a mouse peritonitis model. While 5 of 17 endocarditis isolates were Acm nonproducers and failed to adhere to collagen in vitro, all had an intact, highly conserved acm locus. Highly reduced acm mRNA levels (>or=50-fold reduction relative to an Acm producer) were found in three of these five nonadherent isolates, including the sequenced strain TX0016, by quantitative reverse transcription-PCR, indicating that acm transcription is downregulated in vitro in these isolates. However, examination of TX0016 cells obtained directly from infected rat vegetations by flow cytometry showed that Acm was present on 40% of cells grown during infection. Finally, we demonstrated a significant reduction in E. faecium collagen adherence by affinity-purified anti-Acm antibodies from E. faecium endocarditis patient sera, suggesting that Acm may be a potential immunotarget for strategies to control this emerging pathogen.
authors.affiliation: Department of Internal Medicine, Division of Infectious Diseases, Center for the Study of Emerging and Re-Emerging Pathogens, University of Texas Medical School, Houston, Texas 77030, USA.
authors.forename: Sreedhar R
authors.initials: SR
authors.lastname: Nallapareddy
authors.forename: Kavindra V
authors.initials: KV
authors.lastname: Singh
authors.forename: Barbara E
authors.initials: BE
authors.lastname: Murray
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001422:Bacterial Adhesion; D003094:Collagen / Q000378:metabolism; D004697:Endocarditis, Bacterial / Q000382:microbiology*; D016984:Enterococcus faecium / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology*; D006801:Humans; D051379:Mice; D016254:Mutagenesis, Insertional; D016019:Survival Analysis; D037521:Virulence Factors / Q000235:genetics / Q000502:physiology*
chemical_list: C473793:Acm protein, Enterococcus faecium; D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D037521:Virulence Factors; D003094:Collagen