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Infection and Immunity, June 2006, p. 3415-3426, Vol. 74, No. 6
0019-9567/06/$08.00+0     doi:10.1128/IAI.00392-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of Staphylococcus aureus Proteins Recognized by the Antibody-Mediated Immune Response to a Biofilm Infection

Rebecca A. Brady,1 Jeff G. Leid,2 Anne K. Camper,3 J. William Costerton,4 and Mark E. Shirtliff1,5*

Department of Microbiology and Immunology, University of Maryland—Baltimore, School of Medicine, 660 W. Redwood Street, Baltimore, Maryland 21201,1 Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona 86011,2 Center for Biofilm Engineering, Montana State University, 366 EPS Building, PO Box 173980, Bozeman, Montana 59717,3 Center for Biofilms, School of Dentistry, University of Southern California, Room 4360 DEN, 925 West 34th Street, Los Angeles, California 90089,4 Department of Biomedical Sciences, University of Maryland—Baltimore, Dental School, 666 W. Baltimore Street, Baltimore, Maryland5

Received 10 March 2006/ Returned for modification 23 March 2006/ Accepted 28 March 2006

Staphylococcus aureus causes persistent, recurrent infections (e.g., osteomyelitis) by forming biofilms. To survey the antibody-mediated immune response and identify those proteins that are immunogenic in an S. aureus biofilm infection, the tibias of rabbits were infected with methicillin-resistant S. aureus to produce chronic osteomyelitis. Sera were collected prior to infection and at 14, 28, and 42 days postinfection. The sera were used to perform Western blot assays on total protein from biofilm grown in vitro and separated by two-dimensional gel electrophoresis. Those proteins recognized by host antibodies in the harvested sera were identified via matrix-assisted laser desorption ionization-time of flight analysis. Using protein from mechanically disrupted total and fractionated biofilm protein samples, we identified 26 and 22 immunogens, respectively. These included a cell surface-associated ß-lactamase, lipoprotein, lipase, autolysin, and an ABC transporter lipoprotein. Studies were also performed using microarray analyses and confirmed the biofilm-specific up-regulation of most of these genes. Therefore, although the biofilm antigens are recognized by the immune system, the biofilm infection can persist. However, these proteins, when delivered as vaccines, may be important in directing the immune system toward an early and effective antibody-mediated response to prevent chronic S. aureus infections. Previous works have identified S. aureus proteins that are immunogenic during acute infections, such as sepsis. However, this is the first work to identify these immunogens during chronic S. aureus biofilm infections and to simultaneously show the global relationship between the antigens expressed during an in vivo infection and the corresponding in vitro transcriptomic and proteomic gene expression levels.

* Corresponding author. Mailing address: Department of Biomedical Sciences, Dental School, University of Maryland—Baltimore, 666 W. Baltimore Street, Rm. 4-G-11, Baltimore, MD 21201. Phone: (410) 706-2263. Fax: (410) 706-0865. E-mail: mshirtliff{at}umaryland.edu.

Editor: J. T. Barbieri

Infection and Immunity, June 2006, p. 3415-3426, Vol. 74, No. 6
0019-9567/06/$08.00+0     doi:10.1128/IAI.00392-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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Copyright © 2006 by the American Society for Microbiology. All rights reserved.