title: The modA10 phasevarion of nontypeable Haemophilus influenzae R2866 regulates multiple virulence-associated traits.
abstract: Non-typeable Haemophilus influenzae (NTHi) is a human restricted commensal and pathogen that elicits inflammation by adhering to and invading airway epithelia cells: transcytosis across these cells can result in systemic infection. NTHi strain R2866 was isolated from the blood of a normal 30-month old infant with meningitis, and is unusual for NTHi in that it is able to cause systemic infection. Strain R2866 is able to replicate in normal human serum due to expression of lgtC which mimics human blood group p(k). R2866 contains a phase-variable DNA methyltransferase, modA10 which switches ON and OFF randomly and reversibly due to polymerase slippage over a long tetrameric repeat tract located in its open reading frame. Random gain or loss of repeats during replication can results in expressed (ON), or not expressed (OFF) states, the latter due to a frameshift or transcriptional termination at a premature stop codon. We sought to determine if the unusual virulence of R2866 was modified by modA10 phase-variation. A modA10 knockout mutant was found to have increased adherence to, and invasion of, human ear and airway monolayers in culture, and increased invasion and transcytosis of polarized human bronchial epithelial cells. Intriguingly, the rate of bacteremia was lower in the infant rat model of infection than a wild-type R2866 strain, but the fatality rate was greater. Transcriptional analysis comparing the modA10 knockout to the R2866 wild-type parent strain showed increased expression of genes in the modA10 knockout whose products mediate cellular adherence. We conclude that loss of ModA10 function in strain R2866 enhances colonization and invasion by increasing expression of genes that allow for increased adherence, which can contribute to the increased virulence of this strain.
authors.affiliation: Department of Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, 73104, OK, USA.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: VanWagoner
authors.affiliation: Institute for Glycomics, Griffith University, Gold Coast, Queensland, 4222, Australia.
authors.forename: John M
authors.initials: JM
authors.lastname: Atack
authors.affiliation: Center for Global Infectious Disease Research, Seattle Children's Hospital, Seattle, WA, 98105, USA.
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Nelson
authors.affiliation: Center for Global Infectious Disease Research, Seattle Children's Hospital, Seattle, WA, 98105, USA.
authors.forename: Hannah K
authors.initials: HK
authors.lastname: Smith
authors.affiliation: School of Chemical and Molecular Bioscience, University of Queensland, St. Lucia, Brisbane, Queensland, 4072, Australia.
authors.forename: Kate L
authors.initials: KL
authors.lastname: Fox
authors.affiliation: Institute for Glycomics, Griffith University, Gold Coast, Queensland, 4222, Australia.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Jennings
authors.affiliation: Department of Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, 73104, OK, USA.
authors.forename: Terrence L
authors.initials: TL
authors.lastname: Stull
authors.affiliation: Center for Global Infectious Disease Research, Seattle Children's Hospital, Seattle, WA, 98105, USA. Electronic address: arnold.smith@seattlechildrens.org.
authors.forename: Arnold L
authors.initials: AL
authors.lastname: Smith
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D004195:Disease Models, Animal; D004847:Epithelial Cells; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D006192:Haemophilus Infections / Q000382:microbiology* / Q000401:mortality; D006193:Haemophilus influenzae / Q000472:pathogenicity* / Q000502:physiology*; D006801:Humans; D019655:Quantitative Trait, Heritable; D051381:Rats; D057900:Transcytosis / Q000276:immunology; D014774:Virulence
chemical_list: D001426:Bacterial Proteins
keywords: DNA methylase; Haemophilus influenzae; Pathogenesis; Phase variation; Virulence