Summary: | <p>Abstract</p> <p>Background</p> <p><it>Streptococcus gallolyticus </it>subsp. <it>gallolyticus </it>is an important causative agent of infective endocarditis (IE) but the knowledge on virulence factors is limited and the pathogenesis of the infection is poorly understood. In the present study, we established an experimental <it>in vitro </it>IE cell culture model using EA.hy926 and HUVEC cells to investigate the adhesion and invasion characteristics of 23 <it>Streptococcus gallolyticus </it>subsp. <it>gallolyticus </it>strains from different origins (human IE-derived isolates, other human clinical isolates, animal isolates). Adhesion to eight components of the extracellular matrix (ECM) and the ability to form biofilms <it>in vitro </it>was examined in order to reveal features of <it>S. gallolyticus </it>subsp. <it>gallolyticus </it>endothelial infection. In addition, the strains were analyzed for the presence of the three virulence factors <it>gtf</it>, <it>pilB</it>, and <it>fimB </it>by PCR.</p> <p>Results</p> <p>The adherence to and invasion characteristics of the examined <it>S. gallolyticus </it>subsp. <it>gallolyticus </it>strains to the endothelial cell line EA.hy926 differ significantly among themselves. In contrast, the usage of three different <it>in vitro </it>models (EA.hy926 cells, primary endothelial cells (HUVECs), mechanical stretched cells) revealed no differences regarding the adherence to and invasion characteristics of different strains. Adherence to the ECM proteins collagen I, II and IV revealed the highest values, followed by fibrinogen, tenascin and laminin. Moreover, a strong correlation was observed in binding to these proteins by the analyzed strains. All strains show the capability to adhere to polystyrole surfaces and form biofilms. We further confirmed the presence of the genes of two known virulence factors (<it>fimB</it>: all strains, <it>gtf</it>: 19 of 23 strains) and demonstrated the presence of the gene of one new putative virulence factor (<it>pilB</it>: 9 of 23 strains) by PCR.</p> <p>Conclusion</p> <p>Our study provides the first description of <it>S. gallolyticus </it>subsp. <it>gallolyticus </it>adhesion and invasion of human endothelial cells, revealing important initial information of strain variability, behaviour and characteristics of this as yet barely analyzed pathogen.</p>
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