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JHC exPRESS: First Published March 20, 2006. doi:10.1369/jhc.6A6922.2006
Copyright © Histochemical Society, Inc.

A more recent version of this article appeared on July 1, 2006.
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Distribution of Bacterial Proteins in Biofilms Formed by Non-typeable Haemophilus Influenzae

Paul Webster 1*, Siva Wu 1, Gerardo Gomez 1, Michael Apicella 1, Andrew G. Plaut 1 and Joseph W. St. Geme III1

1 Ahmanson Advanced Electron Microscopy and Imaging Center, House Ear Institute, Los Angeles, California (PW,SW,GG); Department of Microbiology, University of Iowa, Iowa City, Iowa (MA); Gastroenterology Research, Tufts-New England Medical Center, Boston, Massachusetts (AGP); and Department of Pediatrics, Duke University Medical Center, Durham, North Carolina (JWSGIII)

* To whom correspondence should be addressed. E-mail: pwebster{at}hei.org.

Submitted on January 11, 2006
Accepted on 3 March 2006


  Abstract
The ability to preserve the fragile ultrastructural organization of bacterial biofilms using cryo-preparation methods for electron microscopy has enabled us to probe sections through NTHi biofilms and determine the localization of NTHi-specific LOS and proteins within these structures. Some of the proteins we examined are currently being considered as candidates for vaccine development so it is important that their distribution and accessibility within the biofilms formed by NTHi be determined. We have localized lipooligosaccharide (LOS) to the extracellular matrix of the biofilm and the P6 outer membrane protein to the membrane of what appear to be viable bacteria within the biofilm. The Hap and HWM1/HMW2 adhesive proteins were associated with bacteria within the biofilm and were present in the biofilm extracellular matrix. The IgA1 protease is a secreted protein that was also associated with NTHi in the biofilm and was in the extracellular matrix, but was more concentrated in the top region of the biofilm, suggesting a role in protecting biofilm bacteria from antibody attack.

Key Words: OMP P6, HMW proteins, Hap protein, IgA1 protease, electron microscopy, extracellular matrix, immunocytochemistry, stereology


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