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Infection and Immunity, August 2002, p. 4661-4668, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4661-4668.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Acylation of the Lipooligosaccharide of Haemophilus influenzae and Colonization: an htrB Mutation Diminishes the Colonization of Human Airway Epithelial Cells

W. Edward Swords,1,{dagger} Deborah L. Chance,2 Leah A. Cohn,3 Jianqiang Shao,1 Michael A. Apicella,1 and Arnold L. Smith2*

Department of Microbiology, University of Iowa, Iowa City, Iowa,1 Department of Molecular Microbiology and Immunology, School of Medicine,2 Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri-Columbia, Columbia, Missouri3

Received 26 December 2001/ Returned for modification 27 February 2002/ Accepted 2 May 2002

Haemophilus influenzae is a commensal and opportunistic pathogen of the human airways. A number of surface molecules contribute to colonization of the airways by H. influenzae, such as adhesins, including structures found in the lipooligosaccharide (LOS). A human bronchiolar xenograft model was employed to investigate the host-bacterial interactions involved in the colonization of the airway by H. influenzae. Differential display was used to identify H. influenzae mRNA that reflect genes which were preferentially expressed in the xenograft compared to growth. Eleven mRNA fragments had consistent increased expression when the bacteria grew in xenografts. On sequencing these fragments, eight open reading frames were identified. Three of these had no match in the NCBI or the TIGR database, while an additional three were homologous to genes involved in heme or iron acquisition and utilization: two of the mRNAs encoded proteins homologous to enzymes involved in LOS biosynthesis: a heptosyl transferase (rfaF) involved in the synthesis of the LOS core and a ketodeoxyoctonate phosphate-dependent acyltransferase (htrB) that performs one of the late acylation reactions in lipid A synthesis. Inoculation of human bronchiolar xenografts revealed a significant reduction in colonization capacity by htrB mutants. In vitro, htrB mutants elicited lesser degrees of cytoskeletal rearrangement and less stimulation of host cell signaling with 16HBE14o- cells and decreased intracellular survival. These results implicate acylation of H. influenzae lipid A as playing a key role in the organisms' colonization of the normal airway.

* Corresponding author. Mailing address: Department of Molecular Microbiology and Immunology, M616 Medical Sciences Bldg., DC044.00, Columbia, MO 65212. Phone: (573) 882-8989. Fax: (573) 882-4287. E-mail: smithal{at}health.missouri.edu.

Editor: B. B. Finlay

{dagger} Present address: Department of Microbiology and Immunology, Wake Forest University School of Medicine, Winston-Salem, N.C.

Infection and Immunity, August 2002, p. 4661-4668, Vol. 70, No. 8
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.8.4661-4668.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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