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Journal of Bacteriology, July 2004, p. 4209-4217, Vol. 186, No. 13
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.13.4209-4217.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Evolutionary and Functional Relationships among the Nontypeable Haemophilus influenzae HMW Family of Adhesins{dagger},{ddagger}

Amy Z. Buscher,1 Katie Burmeister,1 Stephen J. Barenkamp,2 and Joseph W. St. Geme III1*

Edward Mallinckrodt Department of Pediatrics and Department of Molecular Microbiology, Washington University School of Medicine,1 Department of Pediatrics, St. Louis University School of Medicine, and The Pediatric Research Institute, Cardinal Glennon Children's Hospital, St. Louis, Missouri2

Received 15 December 2003/ Accepted 12 March 2004

Nontypeable Haemophilus influenzae (NTHi) is a common cause of localized respiratory tract disease and initiates infection by colonizing the nasopharynx. Approximately 75 to 80% of NTHi clinical isolates produce proteins that belong to the HMW family of adhesins, which are believed to facilitate colonization. The prototype HMW adhesins are designated HMW1 and HMW2 and were identified in NTHi strain 12. HMW1 and HMW2 are 71% identical and 80% similar overall, yet display differing cellular binding specificities. In the present study we set out to define more clearly the relationships between HMW1 and HMW2 and other members of the HMW family of adhesins. PCR analysis of 49 epidemiologically distinct isolates revealed that all strains possessing hmw genes as determined by Southern analysis contain two hmw loci in conserved, unlinked physical locations on the chromosome. Functional analysis of the HMW adhesins produced by three unrelated strains demonstrated that each isolate possesses one protein with HMW1-like adherence properties and another with HMW2-like adherence properties. These findings suggest that the hmw1 and hmw2 loci may have arisen via a gene duplication event in an ancestral strain. In addition, they support the hypothesis that the distinct binding specificities of HMW1 and HMW2 emerged early and have persisted over time, suggesting an ongoing selective advantage.

* Corresponding author. Mailing address: Department of Pediatrics, Washington University Medical School, 660 S. Euclid Ave., Campus Box 8208, St. Louis, MO 63110. Phone: 314-286-2887. Fax: 314-286-2895. E-mail: stgeme{at}borcim.wustl.edu.

{dagger} Supplemental material for this article may be found at http://jb.asm.org.

{ddagger} Dedicated to the memory of Katie Burmeister.

Journal of Bacteriology, July 2004, p. 4209-4217, Vol. 186, No. 13
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.13.4209-4217.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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