Identification of plasma proteins adsorbed on hemodialysis tubing that promote Staphylococcus aureus adhesion

François, Patrice; Schrenzel, Jacques; Stoerman-Chopard, Catherine; Favre, H; Herrmann, Mathias; Foster, Timothy J.; Lew, Daniel Pablo; Vaudaux, Pierre

doi:10.1016/s0022-2143(00)70018-7

Cited by 69 publications

(53 citation statements)

References 62 publications

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“…Aliquots (0.5 ml) were monitored for protein content by spectrometry (280 nm) and for FITC content by fluorometry (excitation, 485 nm; emission, 515 nm). Fractions containing the fluorescent protein were pooled, and the protein content was determined by the Pierce method (2,15).…”

Section: Reagentsmentioning

confidence: 99%

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Heterologously Expressed Staphylococcus aureus Fibronectin-Binding Proteins Are Sufficient for Invasion of Host Cells

et al. 2000

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Staphylococcus aureus is generally considered an extracellular pathogen (12, 44). However, it was shown in vitro over several years that S. aureus can invade a variety of nonprofessional phagocytes of different species. Many studies have used cells of nonhuman origin, mainly bovine endothelial cells. S. aureus invasion of human cells has been described so far for endothelial cells (8,38), epithelial cells (7,47,50), osteoblasts (24), and fibroblasts (50).S. aureus is among the most important bacterial pathogens for humans. It commonly causes superficial skin infections. Upon bloodstream dissemination or by continuous spread, it can readily survive in various deep tissues and cause, among others, abscess formation, osteomyelitis, endocarditis, and sepsis. Since these S. aureus infections are frequently long lasting, are difficult to eradicate, and often relapse after antibiotic treatment, prolonged treatment, including the use of cell-permeating antimicrobial drugs, is often required (30, 31). Invasion of nonprofessional phagocytes may provide a protected niche for S. aureus, but the precise role of invasion of host cells in S. aureus pathology in vivo is still not known.We have previously analyzed the process of cellular S. aureus invasion and elucidated the molecular mechanism of the internalization step (50). Bacterial entry proceeds by a "zippertype" mechanism, which resembles professional phagocytosis, and requires bacterial surface proteins, namely, adhesins (also termed microbial surface components recognizing adhesive matrix molecules [MSCRAMMs] [13]). The internalization process if F actin dependent, as demonstrated by complete inhibition by cytochalasin D. By use of isogenic deletion mutants, we identified fibronectin-binding protein (FnBP) as the necessary S. aureus invasin. Using competition experiments, we mapped the region of interaction with the host cell to the major fibronectin-binding domain of the FnBP, the D1 to D4 repeats. Fibronectin acts as a bridging molecule between the FnBP and the fibronectin receptor integrin ␣ 5 ␤ 1 , an adhesion molecule on the host cell. Cellular invasion is highly conserved in clinical S. aureus isolates (15 out of 15 tested were invasive), and the mechanism elucidated for the epithelial cell model of 293 cells is also valid for ECV304 cells, skin fibroblasts (50), and human umbilical vein endothelial cells (B. Sinha, unpublished data). These data are compatible with the results obtained by four different groups, who found that FnBPs are essential either for adhesion to or invasion of human endothelial cells (42), bovine mammary epithelial cells (9, 27), and mouse fibroblastic cells (14).Our previous data did not support a role for S. aureus invasins other than FnBPs but could not exclude necessary (S. aureus-specific) coreceptors. However, they largely excluded an efficient second invasin in S. aureus (50), unlike the situation found for streptococcal M proteins. This study was performed to address the questions of whether S. aureus FnBPs are sufficient to confer...

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Section: Reagentsmentioning

confidence: 99%

“…Fibronectin binding was determined with formaldehyde-fixed late-logarithmic-phase bacteria and FITClabeled soluble fibronectin (5 g/ml) in a flow cytometric assay as previously described (15). Incubation was performed on an inverting shaker for 2 h at 4°C.…”

Section: Reagentsmentioning

confidence: 99%

Heterologously Expressed Staphylococcus aureus Fibronectin-Binding Proteins Are Sufficient for Invasion of Host Cells

et al. 2000

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“…In addition, strain Newman strongly expresses Eap (extracellular adherence protein), which among other ligands binds fibronectin (20), thus rendering the interpretation of data derived from in vivo (4, 5, 8, 23-25, 39, 44, 50, 58, 59) and in vitro (6,9,12,13,16,20,32,34,42,43,48,62,66,67,69,70) studies rather difficult.…”

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Truncation of Fibronectin-Binding Proteins in Staphylococcus aureus Strain Newman Leads to Deficient Adherence and Host Cell Invasion Due to Loss of the Cell Wall Anchor Function

et al. 2004

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Staphylococcus aureus fibronectin-binding proteins (FnBPs) play a critical role in S. aureus pathogenesis. FnBPs mediate adhesion to fibronectin and invasion of mammalian cells, including epithelial, endothelial, and fibroblastic cells, by fibronectin bridging to the host cell fibronectin receptor integrin (␣ 5 )␤ 1 . Strain Newman is a laboratory strain frequently used for genetic, functional, and in vivo studies. However, despite pronounced production of FnBPs, strain Newman is only weakly adherent to immobilized Fn and weakly invasive. We examined whether these effects are due to a structural difference of FnBPs. Here, we show that both fnbA Newman and fnbB Newman contain a centrally located point mutation resulting in a stop codon. This leads to a truncation of both FnBPs at the end of the C domain at identical positions. Most likely, the stop codon occurred first in fnbB Newman and was subsequently transferred to fnbA Newman by replacement of the entire region encompassing the C, D, and W domains with the respective sequence of fnbB Newman . Using heterologous expression in Staphylococcus carnosus, we found that truncated FnBPs were completely secreted into the culture medium and not anchored to the cell wall, since they lack the sortase motif (LPETG). Consequently, this led to a loss of FnBP-dependent functions, such as strong adhesion to immobilized fibronectin, binding of fibrinogen, and host cell invasion. This mutation may explain some of the earlier reported conflicting data with strain Newman. Thus, care should be taken when drawing negative conclusions about the role of FnBPs as a virulence factor in a given model. Staphylococcus aureus fibronectin-binding proteins (FnBPs)appear to play a critical role in S. aureus pathogenesis (35), as has been shown, e.g., for infective endocarditis (29, 49) and osteomyelitis (22). However, so far, often conflicting results have been reported that argue for (3,22,29,38,49) or against (2, 5, 10, 33, 41) their role as virulence factors in a given model in vivo. In most situations, the adhesive and invasive functions of FnBPs are the likely functional link in pathogenicity. Adhesion to fibronectin and invasion of mammalian cells, including epithelial, endothelial, and fibroblastic cells, depend on fibronectin bridging between FnBPs and the host fibronectin receptor integrin (␣ 5 )␤ 1 (54).During the analysis of the mechanism for cellular invasion of S. aureus, we found that several reference strains are invasion deficient compared to clinical isolates (54). Strain Newman is a laboratory strain frequently used for genetic, functional, and in vivo studies. Despite strong production of FnBPs (57), strain Newman is only weakly adherent to immobilized Fn and weakly invasive (54). Another often-used reference strain, 8325-4, is also only weakly invasive (54); however, this appears to be due to regulatory defects resulting in low FnBP expression rather than structural FnBP modifications. Strain 8325-4 has a regulatory defect (rsbU) (28) and expresses FnBPs at a low level (6...

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“…It is a common etiologic agent in neonatal septicemia and in peritonitis, causing mainly foreign body-associated infections. In patients, proteins such as albumin, fibrinogen, fibronectin, and von Willebrand factor rapidly cover implants (3). S. epidermidis possesses an unknown number of proteins that contribute to specific adhesion to these molecules.…”

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The Fibrinogen Binding Protein of Staphylococcus epidermidis Is a Target for Opsonic Antibodies

2004

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Antibodies against the fibrinogen binding protein (Fbe) of Staphylococcus epidermidis significantly increased macrophage phagocytosis. Antibodies against autolysin E were opsonic but to a lesser extent. Antibodies against a novel, putatively surface-located antigen were unable to enhance phagocytosis. The severity of systemic infection of mice with S. epidermidis was reduced if the bacteria were preopsonized with anti-Fbe prior to administration. Fbe is thus a strong candidate for protein vaccination against S. epidermidis infection, and antibodies against Fbe can be used to prevent or treat infections caused by S. epidermidis.

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Identification of plasma proteins adsorbed on hemodialysis tubing that promote Staphylococcus aureus adhesion

Cited by 69 publications

References 62 publications

Heterologously Expressed Staphylococcus aureus Fibronectin-Binding Proteins Are Sufficient for Invasion of Host Cells

Heterologously Expressed Staphylococcus aureus Fibronectin-Binding Proteins Are Sufficient for Invasion of Host Cells

Truncation of Fibronectin-Binding Proteins in Staphylococcus aureus Strain Newman Leads to Deficient Adherence and Host Cell Invasion Due to Loss of the Cell Wall Anchor Function

The Fibrinogen Binding Protein of Staphylococcus epidermidis Is a Target for Opsonic Antibodies

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