Bacillus anthracis Protease InhA Increases Blood-Brain Barrier Permeability and Contributes to Cerebral Hemorrhages

Hemorrhagic meningitis is a fatal complication of anthrax, but its pathogenesis remains poorly understood. The present study examined the role of B. anthracis-secreted metalloprotease InhA on monolayer integrity and permeability of human brain microvasculature endothelial cells (HBMECs) which constitute the blood-brain barrier (BBB). Treatment of HBMECs with purified InhA resulted in a time-dependent decrease in trans-endothelial electrical resistance (TEER) accompanied by zonula occluden-1 (ZO-1) degradation. An InhA-expressing B. subtilis exhibited increased permeability of HBMECs, which did not occur with the isogenic inhA deletion mutant (ΔinhA) of B. anthracis, compared with the corresponding wild-type strain. Mice intravenously administered with purified InhA or nanoparticles-conjugated to InhA demonstrated a time-dependent Evans Blue dye extravasation, leptomeningeal thickening, leukocyte infiltration, and brain parenchymal distribution of InhA indicating BBB leakage and cerebral hemorrhage. Mice challenged with vegetative bacteria of the ΔinhA strain of B. anthracis exhibited a significant decrease in leptomeningeal thickening compared to the wildtype strain. Cumulatively, these findings indicate that InhA contributes to BBB disruption associated with anthrax meningitis through proteolytic attack on the endothelial tight junctional protein zonula occluden (ZO)-1.

• Publication year

  2011

• Venue

  PLoS ONE

• Publication date

  2011-03-17

• Fields of study

  Biology, Medicine, Environmental Science

• Identifiers
  DOI 10.1371/journal.pone.0017921PMID 21437287PMCID 3060093
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• Bacillus anthracis lacking inhA shows reduced leptomeningeal thickening after vegetative-bacteria challenge compared with the wild-type strain.

  Confidence 0.95

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• Intravenous purified InhA or nanoparticle-conjugated InhA produces Evans Blue dye extravasation, leptomeningeal thickening, leukocyte infiltration, and brain parenchymal distribution of InhA in mice.

  Confidence 0.97

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• An InhA-expressing Bacillus subtilis increases permeability of human brain microvasculature endothelial cells, whereas the isogenic ΔinhA mutant does not show the same effect relative to wild-type Bacillus anthracis.

  Confidence 0.96

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• InhA treatment decreases trans-endothelial electrical resistance and degrades zonula occluden-1 in human brain microvasculature endothelial cells, consistent with impaired barrier integrity.

  Confidence 0.98

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review

• blood-brain barrier

  biological barrier, physiological barrier

  The endothelial barrier that restricts passage between the bloodstream and brain tissue.

  Aliases: BBB

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• cerebral hemorrhage

  condition, pathology

  Bleeding within the brain or its coverings, used here to describe the pathological outcome associated with barrier disruption.

  Aliases: brain hemorrhage, hemorrhage in the brain

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• evans blue dye extravasation

  measurement, permeability assay

  Leakage of intravenously administered Evans Blue dye into tissues used as a readout of vascular permeability.

  Aliases: Evans Blue extravasation, EB extravasation

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• human brain microvasculature endothelial cells

  cell line, endothelial cells

  Human brain endothelial cells used here as an in vitro model of the blood-brain barrier.

  Aliases: HBMECs

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• inha

  protein, metalloprotease

  A Bacillus anthracis-secreted metalloprotease examined for its effects on endothelial barrier integrity and brain vascular leakage.

  Aliases: InhA protease, B. anthracis InhA

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• zonula occluden-1 (zo-1)

  protein, tight junction protein

  A tight-junction scaffold protein in endothelial monolayers that helps maintain barrier structure.

  Aliases: ZO-1, zonula occludin-1

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review
• δinha strain

  bacterial strain, mutant strain

  An isogenic Bacillus anthracis mutant in which the inhA gene has been deleted.

  Aliases: inhA deletion mutant, ΔinhA mutant

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewB (s683577b42) reviewAK (4715169a40) review

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