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Infection and Immunity, February 2007, p. 958-965, Vol. 75, No. 2
0019-9567/07/$08.00+0     doi:10.1128/IAI.01691-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Phosphorylcholine Decreases Early Inflammation and Promotes the Establishment of Stable Biofilm Communities of Nontypeable Haemophilus influenzae Strain 86-028NP in a Chinchilla Model of Otitis Media{triangledown} ,{dagger}

Wenzhou Hong,1,{ddagger} Kevin Mason,2,{ddagger} Joseph Jurcisek,2 Laura Novotny,2 Lauren O. Bakaletz,2 and W. Edward Swords1*

Department of Microbiology and Immunology, Wake Forest University Health Sciences, Winston-Salem, North Carolina,1 Center for Microbial Pathogenesis, Columbus Childrens' Research Institute, Columbus, Ohio2

Received 23 October 2006/ Returned for modification 8 November 2006/ Accepted 16 November 2006

Nontypeable Haemophilus influenzae (NTHi) is a leading causative agent of otitis media. Much of the inflammation occurring during NTHi disease is initiated by lipooligosaccharides (LOS) on the bacterial surface. Phosphorylcholine (PCho) is added to some LOS forms in a phase-variable manner, and these PCho+ variants predominate in vivo. Thus, we asked whether this modification confers some advantage during infection. Virulence of an otitis media isolate (NTHi strain 86-028NP) was compared with that of an isogenic PCho transferase (licD) mutant using a chinchilla (Chinchilla lanigera) model of otitis media. Animals infected with NTHi 86-028NP licD demonstrated increased early inflammation and a delayed increase in bacterial counts compared to animals infected with NTHi 86-028NP. LOS purified from chinchilla-passed NTHi 86-028NP had increased PCho content compared to LOS purified from the inoculum. Both strains were recovered from middle ear fluids as long as 14 days postinfection. Biofilms were macroscopically visible in the middle ears of euthanized animals infected with NTHi 86-028NP 7 days and 14 days postchallenge. Conversely, less dense biofilms were observed in animals infected with NTHi 86-028NP licD 7 days postinfection, and none of the animals infected with NTHi 86-028NP licD had a visible biofilm by 14 days. Fluorescent antibody staining revealed PCho+ variants within biofilms, similar to our prior results with tissue culture cells in vitro (S. L. West-Barnette, A. Rockel, and W. E. Swords, Infect. Immun. 74:1828-1836, 2006). Animals coinfected with equal proportions of both strains had equal persistence of each strain and somewhat greater severity of disease. We thus conclude that PCho promotes NTHi infection and persistence by reducing the host inflammatory response and by promoting formation of stable biofilm communities.

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

{triangledown} Published ahead of print on 27 November 2006.

Editor: J. N. Weiser

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

{ddagger} These authors contributed equally to this work.

Infection and Immunity, February 2007, p. 958-965, Vol. 75, No. 2
0019-9567/07/$08.00+0     doi:10.1128/IAI.01691-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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