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Infection and Immunity, January 2004, p. 106-113, Vol. 72, No. 1
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.1.106-113.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Sialylation of Lipooligosaccharides Promotes Biofilm Formation by Nontypeable Haemophilus influenzae

W. Edward Swords,1* Miranda L. Moore,1 Luciana Godzicki,2 Gail Bukofzer,2 Michael J. Mitten,2 and Jessica VonCannon1

Department of Microbiology and Immunology, Wake Forest University Health Sciences, Winston-Salem, North Carolina,1 Discovery and Clinical Microbiology, Abbott Laboratories, Abbott Park, Illinois2

Received 30 July 2003/ Returned for modification 5 September 2003/ Accepted 24 September 2003

Nontypeable Haemophilus influenzae (NTHi) is a major cause of opportunistic respiratory tract infections, including otitis media and bronchitis. The persistence of NTHi in vivo is thought to involve bacterial persistence in a biofilm community. Therefore, there is a need for further definition of bacterial factors contributing to biofilm formation by NTHi. Like other bacteria inhabiting host mucosal surfaces, NTHi has on its surface a diverse array of lipooligosaccharides (LOS) that influence host-bacterial interactions. In this study, we show that LOS containing sialic (N-acetyl-neuraminic) acid promotes biofilm formation by NTHi in vitro and bacterial persistence within the middle ear or lung in vivo. LOS from NTHi in biofilms was sialylated, as determined by comparison of electrophoretic mobilities and immunochemical reactivities before and after neuraminidase treatment. Biofilm formation was significantly reduced in media lacking sialic acid, and a siaB (CMP-sialic acid synthetase) mutant was deficient in biofilm formation in three different in vitro model systems. The persistence of an asialylated siaB mutant was attenuated in a gerbil middle ear infection model system, as well as in a rat pulmonary challenge model system. These data show that sialylated LOS glycoforms promote biofilm formation by NTHi and persistence in vivo.

* Corresponding author. Mailing address: 5101A Gray Building, Medical Center Blvd., Winston-Salem, NC 27157. Phone: (336) 713-5049. Fax: (336) 716-9928. E-mail: wswords{at}wfubmc.edu.

Editor: J. N. Weiser

Infection and Immunity, January 2004, p. 106-113, Vol. 72, No. 1
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.1.106-113.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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