title: Penetration of the blood-brain barrier by Bacillus anthracis requires the pXO1-encoded BslA protein.
abstract: Anthrax is a zoonotic disease caused by the gram-positive spore-forming bacterium Bacillus anthracis. Human infection occurs after the ingestion, inhalation, or cutaneous inoculation of B. anthracis spores. The subsequent progression of the disease is largely mediated by two native virulence plasmids, pXO1 and pXO2, and is characterized by septicemia, toxemia, and meningitis. In order to produce meningitis, blood-borne bacteria must interact with and breach the blood-brain barrier (BBB) that is composed of a specialized layer of brain microvascular endothelial cells (BMEC). We have recently shown that B. anthracis Sterne is capable of penetrating the BBB in vitro and in vivo, establishing the classic signs of meningitis; however, the molecular mechanisms underlying the central nervous system (CNS) tropism are not known. Here, we show that attachment to and invasion of human BMEC by B. anthracis Sterne is mediated by the pXO1 plasmid and an encoded envelope factor, BslA. The results of studies using complementation analysis, recombinant BslA protein, and heterologous expression demonstrate that BslA is both necessary and sufficient to promote adherence to brain endothelium. Furthermore, mice injected with the BslA-deficient strain exhibited a significant decrease in the frequency of brain infection compared to mice injected with the parental strain. In addition, BslA contributed to BBB breakdown by disrupting tight junction protein ZO-1. Our results identify the pXO1-encoded BslA adhesin as a critical mediator of CNS entry and offer new insights into the pathogenesis of anthrax meningitis.
authors.affiliation: Department of Biology, San Diego State University, San Diego, CA 92182, USA.
authors.forename: Celia M
authors.initials: CM
authors.lastname: Ebrahimi
authors.forename: Justin W
authors.initials: JW
authors.lastname: Kern
authors.forename: Tamsin R
authors.initials: TR
authors.lastname: Sheen
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Ebrahimi-Fardooee
authors.forename: Nina M
authors.initials: NM
authors.lastname: van Sorge
authors.forename: Olaf
authors.initials: O
authors.lastname: Schneewind
authors.forename: Kelly S
authors.initials: KS
authors.lastname: Doran
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics; D000818:Animals; D000881:Anthrax / Q000382:microbiology*; D001408:Bacillus anthracis / Q000235:genetics / Q000472:pathogenicity*; D001422:Bacterial Adhesion / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D001812:Blood-Brain Barrier / Q000382:microbiology*; D002460:Cell Line; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D051379:Mice; D008853:Microscopy; D010957:Plasmids / Q000235:genetics
chemical_list: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins