Staphylococcus aureus Host Cell Invasion and Virulence in Sepsis Is Facilitated by the Multiple Repeats within FnBPA

PLoS Pathogens

Andrew M. Edwards¹  Ruth C. Massey¹  Jennifer R. Potts²  Elisabet Josefsson³ 

[1] Department of Biology and Biochemistry, University of Bath, Bath, United Kingdom;Department of Biology and Department of Chemistry, University of York, York, United Kingdom;Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden

关键词: Staphylococcus aureus;    Lactococcus lactis;    Endothelial cells;    Enzyme-linked immunoassays;    Blood;    Fibrinogen;    Sepsis;    Cell binding;   

DOI  :  10.1371/journal.ppat.1000964

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Entry of Staphylococcus aureus into the bloodstream can lead to metastatic abscess formation and infective endocarditis. Crucial to the development of both these conditions is the interaction of S. aureus with endothelial cells. In vivo and in vitro studies have shown that the staphylococcal invasin FnBPA triggers bacterial invasion of endothelial cells via a process that involves fibronectin (Fn) bridging to α5β1 integrins. The Fn-binding region of FnBPA usually contains 11 non-identical repeats (FnBRs) with differing affinities for Fn, which facilitate the binding of multiple Fn molecules and may promote integrin clustering. We thus hypothesized that multiple repeats are necessary to trigger the invasion of endothelial cells by S. aureus. To test this we constructed variants of fnbA containing various combinations of FnBRs. In vitro assays revealed that endothelial cell invasion can be facilitated by a single high-affinity, but not low-affinity FnBR. Studies using a nisin-inducible system that controlled surface expression of FnBPA revealed that variants encoding fewer FnBRs required higher levels of surface expression to mediate invasion. High expression levels of FnBPA bearing a single low affinity FnBR bound Fn but did not invade, suggesting that FnBPA affinity for Fn is crucial for triggering internalization. In addition, multiple FnBRs increased the speed of internalization, as did higher expression levels of FnBPA, without altering the uptake mechanism. The relevance of these findings to pathogenesis was demonstrated using a murine sepsis model, which showed that multiple FnBRs were required for virulence. In conclusion, multiple FnBRs within FnBPA facilitate efficient Fn adhesion, trigger rapid bacterial uptake and are required for pathogenesis.

【 授权许可】

CC BY