title: Fimbria-mediated enhanced attachment of nontypeable Haemophilus influenzae to respiratory syncytial virus-infected respiratory epithelial cells.
abstract: 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.
authors.affiliation: Department of Pediatrics, University of Texas Medical Branch, Galveston 77555-0371, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Jiang
authors.forename: N
authors.initials: N
authors.lastname: Nagata
authors.forename: E
authors.initials: E
authors.lastname: Molina
authors.forename: L O
authors.initials: LO
authors.lastname: Bakaletz
authors.forename: H
authors.initials: H
authors.lastname: Hawkins
authors.forename: J A
authors.initials: JA
authors.lastname: Patel
mesh_terms: D000956:Antigens, Viral / Q000096:biosynthesis; D001422:Bacterial Adhesion / Q000502:physiology*; D001425:Bacterial Outer Membrane Proteins / Q000502:physiology; D004847:Epithelial Cells / Q000382:microbiology* / Q000648:ultrastructure / Q000821:virology; D010861:Fimbriae, Bacterial / Q000502:physiology* / Q000648:ultrastructure; D006193:Haemophilus influenzae / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D008168:Lung / Q000382:microbiology* / Q000648:ultrastructure / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000276:immunology; D014407:Tumor Cells, Cultured
chemical_list: D000956:Antigens, Viral; D001425:Bacterial Outer Membrane Proteins; C082912:outer membrane protein P5, Haemophilus