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J. Bacteriol. doi:10.1128/JB.01935-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Biofilms formed by nontypeable Haemophilus influenzae in vivo contain both dsDNA as well as type IV pilin protein

Columbus Children's Research Institute, Center for Microbial Pathogenesis, The Ohio State University College of Medicine, 700 Children's Drive, W591, Columbus, OH 43205

^* To whom correspondence should be addressed. Email: BakaletL{at}ccri.net.

	Abstract

Nontypeable Haemophilus influenzae (NTHI) are members of the normal human nasopharyngeal flora, as well as frequent opportunistic pathogens of both the upper and lower respiratory tract. Recently, it's been shown that NTHI can form biofilms both in vitro and in vivo. NTHI within in vitro-formed biofilms differentially express both epitopes of LOS as well as the outer membrane proteins P2, P5 and P6, whereas those generated both in a 96-well plate assay in vitro as well as in a mammalian host, have been shown to incorporate a specific glycoform of sialylated LOS within the biofilm matrix. While DNA has been identified as a key component of the biofilm matrix formed in vitro by several bacterial pathogens, here we demonstrate for the first time that in addition to sialylated LOS, the biofilm formed by NTHI in vivo, contains both type IV pilin protein as well as a significant amount of double-stranded DNA. The DNA appeared to be arranged in a dense interlaced meshwork of fine strands, as well as individual thicker ‘ropes" that span water channels, suggesting that DNA could be imparting structural stability to the biofilm produced by NTHI in vivo. The presence of type IV pilin protein appearing as both small aggregates within the biofilm matrix, as well as tracking along DNA strands support our observations which showed that type IV pili are expressed by NTHI during experimental otitis media when these bacteria form a biofilm in the middle ear space.

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