title: Two distinct pathways for the invasion of Streptococcus pyogenes in non-phagocytic cells.
abstract: Adherence to and invasion of epithelial cells represent important pathogenic mechanisms of Streptococcus pyogenes. A fibronectin-binding surface protein of S. pyogenes, Sfbl protein, has been implicated in both adherence and invasion processes. Invasion of Sfbl-containing strains has been suspected to be responsible for the failure of antibiotics treatment to eradicate S. pyogenes. In this study, we tested the adherence and invasion properties of two well-characterized clinical isolates: A40, which expresses Sfbl; and A8, which is Sfbl negative and is unable to bind fibronectin. In strain A40, Sfbl was the main factor required for attachment and invasion by using fibronectin as a bridging molecule and the alpha5beta1 integrin as cellular receptor. The uptake process was characterized by the generation of large membrane invaginations at the bacteria-cell interface without evidence of actin recruitment or cellular injury. A40 cells were located in phagosomes and, only 24 h after infection, a consistent part of the bacterial population reached the cytoplasm. In contrast, uptake of strain A8 required major rearrangements of cytoskeletal proteins underneath attached bacteria. In A8, a proteinaceous moiety was involved, which does not interact with alpha5beta1 or need any known bridging molecule. Bacterial attachment stimulated elongation and massive recruitment of neighbouring microvilli, which fused to surround streptococcal chains. They led to the generation of large pseudopod-like structures, which engulfed bacteria that were rapidly released and replicated in the cytoplasm. The identification of two completely different uptake pathways reported here provided further evidence regarding the diversity of S. pyogenes isolates and might contribute towards understanding the pathogenesis and persistence of S. pyogenes.
authors.affiliation: Department of Microbial Pathogenicity and Vaccine Research, National Research Centre for Biotechnology, Braunschweig, Germany. gmo@gbf.de
authors.forename: G
authors.initials: G
authors.lastname: Molinari
authors.forename: M
authors.initials: M
authors.lastname: Rohde
authors.forename: C A
authors.initials: CA
authors.lastname: Guzmán
authors.forename: G S
authors.initials: GS
authors.lastname: Chhatwal
mesh_terms: D000199:Actins / Q000378:metabolism; D018829:Adhesins, Bacterial; D001422:Bacterial Adhesion / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000382:microbiology*; D005353:Fibronectins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D013290:Streptococcal Infections / Q000382:microbiology*; D013297:Streptococcus pyogenes / Q000145:classification / Q000472:pathogenicity* / Q000502:physiology
chemical_list: D000199:Actins; D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D005353:Fibronectins; D016023:Integrins; C078635:fibronectin-binding proteins, bacterial