title: Blood-brain barrier invasion by group B Streptococcus depends upon proper cell-surface anchoring of lipoteichoic acid.
abstract: Group B streptococci (GBSs) are the leading cause of neonatal meningitis. GBSs enter the CNS by penetrating the blood-brain barrier (BBB), which consists of specialized human brain microvascular endothelial cells (hBMECs). To identify GBS factors required for BBB penetration, we generated random mutant libraries of a virulent strain and screened for loss of hBMEC invasion in vitro. Two independent hypo-invasive mutants possessed disruptions in the same gene, invasion associated gene (iagA), which encodes a glycosyltransferase homolog. Allelic replacement of iagA in the GBS chromosome produced a 4-fold decrease in hBMEC invasiveness. Mice challenged with the GBS DeltaiagA mutant developed bacteremia comparably to WT mice, yet mortality was significantly lower (20% vs. 90%), as was the incidence of meningitis. The glycolipid diglucosyldiacylglycerol, a cell membrane anchor for lipoteichoic acid (LTA) and predicted product of the IagA glycosyltransferase, was absent in the DeltaiagA mutant, which consequently shed LTA into the media. Attenuation of virulence of the DeltaiagA mutant was found to be independent of TLR2-mediated signaling, but bacterial supernatants from the DeltaiagA mutant containing released LTA inhibited hBMEC invasion by WT GBS. Our data suggest that LTA expression on the GBS surface plays a role in bacterial interaction with BBB endothelium and the pathogenesis of neonatal meningitis.
authors.affiliation: Department of Pediatrics, Division of Infectious Diseases, UCSD School of Medicine, La Jolla, California 92093, USA. kdoran@ucsd.edu
authors.forename: Kelly S
authors.initials: KS
authors.lastname: Doran
authors.forename: Erin J
authors.initials: EJ
authors.lastname: Engelson
authors.forename: Arya
authors.initials: A
authors.lastname: Khosravi
authors.forename: Heather C
authors.initials: HC
authors.lastname: Maisey
authors.forename: Iris
authors.initials: I
authors.lastname: Fedtke
authors.forename: Ozlem
authors.initials: O
authors.lastname: Equils
authors.forename: Kathrin S
authors.initials: KS
authors.lastname: Michelsen
authors.forename: Moshe
authors.initials: M
authors.lastname: Arditi
authors.forename: Andreas
authors.initials: A
authors.lastname: Peschel
authors.forename: Victor
authors.initials: V
authors.lastname: Nizet
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000502:physiology; D001812:Blood-Brain Barrier / Q000378:metabolism / Q000382:microbiology*; D001921:Brain / Q000166:cytology / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D002462:Cell Membrane / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000382:microbiology; D004730:Endothelium, Vascular / Q000166:cytology; D006017:Glycolipids / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D008070:Lipopolysaccharides / Q000378:metabolism*; D016920:Meningitis, Bacterial; D051379:Mice; D008810:Mice, Inbred C57BL; D013292:Streptococcus agalactiae / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity*; D013682:Teichoic Acids / Q000378:metabolism*
chemical_list: D006017:Glycolipids; D008070:Lipopolysaccharides; D013682:Teichoic Acids; C009900:lipoteichoic acid