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Infection and Immunity, October 2004, p. 5668-5675, Vol. 72, No. 10
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.10.5668-5675.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Internalization of Staphylococcus aureus by Human Keratinocytes

Sompid Kintarak,1 Simon A. Whawell,2 Paul M. Speight,2 Samantha Packer,3 and Sean P. Nair3*

Department of Stomatology, Faculty of Dentistry, Prince of Songkla University, Hat-Yai, Songkhla, Thailand,1 Department of Oral Pathology, School of Clinical Dentistry, University of Sheffield, Sheffield,2 Division of Microbial Diseases, Eastman Dental Institute, University College London, London, United Kingdom3

Received 26 February 2004/ Returned for modification 24 April 2004/ Accepted 23 June 2004

Staphylococcus aureus is among the most important human pathogens and causes various superficial and systemic infections. The ability of S. aureus to be internalized by, and survive within, host cells, such as keratinocytes, may contribute to the development of persistent or chronic infections and may finally lead to deeper tissue infections or dissemination. To examine the mechanisms of internalization of S. aureus by keratinocytes, isogenic mutants lacking fibronectin-binding proteins (FnBPs), a recombinant protein consisting of the fibronectin-binding domain of S. aureus FnBPs, and an anti-{alpha}5ß1 antibody were used in cocultures with immortalized keratinocytes and primary keratinocytes. We found that internalization of S. aureus by immortalized keratinocytes requires bacterial FnBPs and is mediated by the major fibronectin-binding integrin {alpha}5ß1. In contrast to internalization by immortalized keratinocytes, internalization of S. aureus by primary keratinocytes could occur through FnBP-dependent and -independent pathways. S. aureus clumping factor B (ClfB), which was recently determined to bind to epithelial cells, was not involved in the uptake of this bacterium by keratinocytes. The identification of an alternate uptake pathway, which is independent of S. aureus FnBPs and host cell {alpha}5ß1, has important implications for the design of therapies targeted to bacterial uptake by host cells.

* Corresponding author. Mailing address: Division of Microbial Diseases, Eastman Dental Institute, University College London, 256 Gray's Inn Road, London WC1X 8LD, United Kingdom. Phone:44(0)207 9151188. Fax: 44(0)207 9151127. E-mail: snair{at}eastman.ucl.ac.uk.

Editor: J. B. Bliska

Infection and Immunity, October 2004, p. 5668-5675, Vol. 72, No. 10
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.10.5668-5675.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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