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Infection and Immunity, October 2002, p. 5339-5345, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5339-5345.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Functional Blocking of Staphylococcus aureus Adhesins following Growth in Ex Vivo Media

Ruth C. Massey,¹ Shobana R. Dissanayeke,¹ Brian Cameron,¹ David Ferguson,¹ Timothy J. Foster,² and Sharon J. Peacock^1*

Nuffield Department of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom,¹ Department of Microbiology, Moyne Institute of Preventive Medicine, Trinity College, Dublin, Ireland²

Received 4 January 2002/ Returned for modification 12 April 2002/ Accepted 20 June 2002

Defining the role of Staphylococcus aureus adhesins in disease pathogenesis may depend on the use of bacteria grown in culture media that more closely reflect the human milieu than conventional broth. This study examined the functional effect on S. aureus adhesins following growth in an ex vivo medium containing a complex mixture of human proteins (used peritoneal dialysate) relative to growth in Todd-Hewitt broth. The adherence of S. aureus, cultured in dialysate, to fibronectin and fibrinogen was markedly reduced despite the expresion of full-length ClfA, ClfB, and fibronectin-binding proteins. Growth in dialysate resulted in the acquisition of a surface coat, as visualized by transmission electron microscopy, which was shown to contain fibronectin, fibrinogen, and immunoglobulins. Adherence of S. aureus to fibrinogen following growth in dialysate was significantly reduced by expression of protein A but was restored following growth in immunoglobulin-depleted dialysate. We conclude that bacterial adherence to solid-phase protein is critically dependent on the culture medium, that S. aureus adhesins may become saturated with target protein prior to contact with solid surfaces, and that there is an interaction between fibrinogen-binding proteins and immunoglobulin bound to protein A following contact with host proteins. These findings have important implications for future studies of S. aureus adhesins.

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* Corresponding author. Mailing address: Department of Microbiology, Level 7, The John Radcliffe Hospital, Headington, Oxford, OX3 9DU, United Kingdom. Phone: 44-1865-220537. Fax: 44-1865-220984. E-mail: sharon.peacock{at}ndcls.ox.ac.uk.

Editor: E. I. Tuomanen

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Infection and Immunity, October 2002, p. 5339-5345, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5339-5345.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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