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Infection and Immunity, January 1999, p. 187-192, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Fimbria-Mediated Enhanced Attachment of Nontypeable Haemophilus influenzae to Respiratory Syncytial Virus-Infected Respiratory Epithelial Cells

Zili Jiang,1 Nobuo Nagata,1,dagger Edgar Molina,1,Dagger Lauren O. Bakaletz,2 Hal Hawkins,3 and Janak A. Patel1,*

Departments of Pediatrics1 and Pathology,3 University of Texas Medical Branch, Galveston, Texas, and Otological Research Laboratories, Department of Otolaryngology, Ohio State University, Columbus, Ohio2

Received 19 August 1998/Returned for modification 17 September 1998/Accepted 8 October 1998

Respiratory syncytial virus (RSV) infection is known to predispose children to otitis media and sinusitis due to bacteria such as nontypeable Haemophilus influenzae (NTHI). In this study, we investigated the role of NTHI surface outer membrane protein P5-homologous fimbriae (P5-fimbriae) in attachment to RSV-exposed A549 epithelial cells. Analysis by fluorescence flow cytometry showed that a live P5-fimbriated NTHI strain (NTHIF+) attached to a higher proportion of RSV-exposed A549 cells than to control cells (mean, 68% for RSV versus 29% for control; P = 0.008), while attachment of the P5-fimbriae-deficient isogenic mutant strain (NTHIF-) was significantly lower than in control cells and rose only slightly following RSV exposure (mean, 17% for RSV versus 10% for control, P = 0.229). Attachment of NTHIF+ did not correlate with the amount of RSV antigen expressed by A549 cells. Furthermore, paraformaldehyde-fixed NTHIF+ also demonstrated an enhanced binding to RSV-exposed cells. Observations by transmission electronic microscopy showed that the mean number of bacteria attached per 100 RSV-exposed A549 cells was higher for NTHIF+ than NTHIF- (99 versus 18; P < 0.001). No intracellular bacteria were identified. UV-irradiated conditioned supernatants collected from RSV-infected A549 cultures (UV-cRSV) also enhanced the attachment of NTHIF+ to A549, suggesting the presence of a preformed soluble mediator(s) in UV-cRSV that enhances the expression of receptors for P5-fimbriae on A549 cells. In summary, RSV infection significantly enhances NTHI attachment to respiratory epithelial cells. P5-fimbria is the critical appendage of NTHI that participates in this attachment. In clinical settings, blocking of the P5-fimbria-mediated attachment of NTHIF+ by passive or active immunity may reduce the morbidity due to NTHI during RSV infection.

* Corresponding author. Mailing address: Division of Pediatric Infectious Diseases, Children's Hospital at the University of Texas Medical Branch, 301 University Blvd., Galveston, TX 77555-0371. Phone: (409) 772-2798. Fax: (409) 747-1753. E-mail: janak.patel{at}utmb.edu.

dagger Present address: Tomakomai City General Hospital, Tomakomai, Japan.

Dagger Present address: Gateway Community Health Center, Laredo, Tex.

Infection and Immunity, January 1999, p. 187-192, Vol. 67, No. 1
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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