Distribution of Bacterial Proteins in Biofilms Formed by Non-typeable Haemophilus influenzae

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 non-typeable Haemophilus influenzae (NTHi) biofilms and determine the localization of NTHi-specific lipooligosacchari...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 2006-07, Vol.54 (7), p.829-842
Main Authors: Webster, Paul, Wu, Siva, Gomez, Gerardo, Apicella, Michael, Plaut, Andrew G, St. Geme, Joseph W., III
Format: Article
Language:English
Subjects:
Adhesins, Bacterial - metabolism
Bacterial Outer Membrane Proteins - metabolism
Bacterial Proteins - metabolism
Bacteriological Techniques
Biofilms
Coloring Agents
Cryopreservation
Gentian Violet
Haemophilus influenzae - isolation & purification
Haemophilus influenzae - metabolism
Haemophilus influenzae - physiology
Haemophilus Vaccines - metabolism
Humans
Immunohistochemistry
Lipopolysaccharides - metabolism
Microscopy, Electron, Scanning
Otitis Media - microbiology
Serine Endopeptidases - metabolism
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Summary: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 non-typeable Haemophilus influenzae (NTHi) biofilms and determine the localization of NTHi-specific lipooligosaccharide (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 LOS to the extracellular matrix (ECM) 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 ECM. The IgA1 protease is a secreted protein that was also associated with NTHi in the biofilm and was in the ECM, but was more concentrated in the top region of the biofilm, suggesting a role in protecting biofilm bacteria from antibody attack.
ISSN:0022-1554
1551-5044
DOI:10.1369/jhc.6A6922.2006