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Applied and Environmental Microbiology, October 2007, p. 6612-6619, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.00855-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Immunoglobulins to Surface-Associated Biofilm Immunogens Provide a Novel Means of Visualization of Methicillin-Resistant Staphylococcus aureus Biofilms{triangledown}

Rebecca A. Brady,1 Jeff G. Leid,2 Jennifer Kofonow,2 J. William Costerton,3 and Mark E. Shirtliff1,4*

Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201,1 Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona 86011,2 School of Dentistry, University of Southern California, Los Angeles, California 90033,3 Department of Biomedical Sciences, University of Maryland Dental School, Baltimore, Maryland 212014

Received 16 April 2007/ Accepted 13 August 2007

Antigens from the methicillin-resistant Staphylococcus aureus (MRSA) cell wall have been shown to be immunogenic in vivo and upregulated during biofilm growth. In this study, we created purified, recombinant forms of selected antigens and biofilm-upregulated, cell wall-associated proteins. These proteins were shown to cause a robust polyclonal immunoglobulin G (IgG) response when used to immunize rabbits. Antibodies against these recombinant proteins bound to the native forms of each protein as harvested from in vitro grown biofilms of MRSA, as determined both via Western blot analysis and immunofluorescence confocal microscopy. These IgGs could be utilized as imaging tools that localize to areas of specific protein production within a biofilm. This work illustrates that immunogenic, cell wall-associated, biofilm-upregulated proteins are promising for in vitro visualization of biofilm growth, architecture, and space-function relationships.

* Corresponding author. Mailing address: Department of Biomedical Sciences, Dental School, University of Maryland—Baltimore, 650 W. Baltimore Street, Room 9414, Baltimore, MD 21201. Phone: (410) 706-2263. Fax: (410) 706-0865. E-mail: mshirtliff{at}umaryland.edu

{triangledown} Published ahead of print on 24 August 2007.

Applied and Environmental Microbiology, October 2007, p. 6612-6619, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.00855-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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