Initial Interactions between Bordetella bronchiseptica and Tracheal Epithelial Cell Cilia

Persistent Link:
http://hdl.handle.net/10150/195708
Title:
Initial Interactions between Bordetella bronchiseptica and Tracheal Epithelial Cell Cilia
Author:
Edwards, Jessica
Issue Date:
2006
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
The Bordetella genus contain primary colonizing bacteria of the mammalian respiratory tract. Bordetella bind specifically to cilia and colonize the healthy conducting airway epithelium. In this dissertation, I examine the following aspects of initial Bordetella/ciliary binding: 1) the contribution of individual Bordetella virulence factors to binding, 2) the identity of ciliary receptors for Bordetella, and 3) the contribution of airway epithelial innate immune effectors in defense against Bordetella colonization. Bordetella/host cell interactions are modeled using Bordetella bronchiseptica and ciliated rabbit tracheal epithelial cells (RTEC) in a 240 second binding assay. B. bronchiseptica expressing the full complement of virulence factors adhere directly to RTEC cilia. Using single knockout strains of B. bronchiseptica in the ciliary binding assay. I show that the virulence factors filamentous hemagglutinin (FHA), fimbriae, pertactin, and adenylate cyclase-hemolysin (CyaA) contributed to ciliary binding, whereas dermonecrotic toxin, FhaS and the type III secretion system did not. Additional B. bronchiseptica adherence factors exist, as a strain that does not express FHA, fimbriae, pertactin or CyaA retained ciliary binding activity. In an attempt to identify ciliary components for binding, B. bronchiseptica bound cilia with greater affinity following enzymatic removal of terminal sialic acid residues from RTEC cilia. Complementary experiments using the immobilized glycocongugates GM1 and asialoGM1 suggest roles for FHA, and possibly fimbriae and pertactin, in ciliary attachment to asialylated compounds. In attempts to identify airway epithelial innate immune responses that limited ciliary attachment, I show that surfactant protein A (SP-A) blocked B. bronchiseptica adherence to RTEC cilia and nitric oxide (NO) inhibited B. bronchiseptica growth. To determine if B. bronchiseptica might encounter NO during colonization, I developed a novel method to isolate ciliated cells from the tracheal mucosa, and showed constitutive expression of iNOS mRNA in these cells. In summary, I present data that further the understanding of initial Bordetella/ciliary interaction and identify potential pathogen and host targets that may be manipulated to alter Bordetella colonization of the airway.
Type:
text; Electronic Dissertation
Degree Name:
PhD
Degree Level:
doctoral
Degree Program:
Cell Biology & Anatomy; Graduate College
Degree Grantor:
University of Arizona
Advisor:
Boitano, Scott
Committee Chair:
Boitano, Scott

Full metadata record

DC FieldValue Language
dc.language.isoENen_US
dc.titleInitial Interactions between Bordetella bronchiseptica and Tracheal Epithelial Cell Ciliaen_US
dc.creatorEdwards, Jessicaen_US
dc.contributor.authorEdwards, Jessicaen_US
dc.date.issued2006en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractThe Bordetella genus contain primary colonizing bacteria of the mammalian respiratory tract. Bordetella bind specifically to cilia and colonize the healthy conducting airway epithelium. In this dissertation, I examine the following aspects of initial Bordetella/ciliary binding: 1) the contribution of individual Bordetella virulence factors to binding, 2) the identity of ciliary receptors for Bordetella, and 3) the contribution of airway epithelial innate immune effectors in defense against Bordetella colonization. Bordetella/host cell interactions are modeled using Bordetella bronchiseptica and ciliated rabbit tracheal epithelial cells (RTEC) in a 240 second binding assay. B. bronchiseptica expressing the full complement of virulence factors adhere directly to RTEC cilia. Using single knockout strains of B. bronchiseptica in the ciliary binding assay. I show that the virulence factors filamentous hemagglutinin (FHA), fimbriae, pertactin, and adenylate cyclase-hemolysin (CyaA) contributed to ciliary binding, whereas dermonecrotic toxin, FhaS and the type III secretion system did not. Additional B. bronchiseptica adherence factors exist, as a strain that does not express FHA, fimbriae, pertactin or CyaA retained ciliary binding activity. In an attempt to identify ciliary components for binding, B. bronchiseptica bound cilia with greater affinity following enzymatic removal of terminal sialic acid residues from RTEC cilia. Complementary experiments using the immobilized glycocongugates GM1 and asialoGM1 suggest roles for FHA, and possibly fimbriae and pertactin, in ciliary attachment to asialylated compounds. In attempts to identify airway epithelial innate immune responses that limited ciliary attachment, I show that surfactant protein A (SP-A) blocked B. bronchiseptica adherence to RTEC cilia and nitric oxide (NO) inhibited B. bronchiseptica growth. To determine if B. bronchiseptica might encounter NO during colonization, I developed a novel method to isolate ciliated cells from the tracheal mucosa, and showed constitutive expression of iNOS mRNA in these cells. In summary, I present data that further the understanding of initial Bordetella/ciliary interaction and identify potential pathogen and host targets that may be manipulated to alter Bordetella colonization of the airway.en_US
dc.typetexten_US
dc.typeElectronic Dissertationen_US
thesis.degree.namePhDen_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineCell Biology & Anatomyen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorBoitano, Scotten_US
dc.contributor.chairBoitano, Scotten_US
dc.contributor.committeememberJost, B. Helenen_US
dc.contributor.committeememberLantz, R. Clarken_US
dc.contributor.committeememberMiesfield, Roger L.en_US
dc.contributor.committeememberWilson, Jean M.en_US
dc.identifier.proquest1499en_US
dc.identifier.oclc659746272en_US
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