scholarly journals Characterization of Innate Immune Responses of Human Endothelial Cells Induced by Porphyromonas gingivalis and Their Derived Outer Membrane Vesicles

2016 ◽  
Vol 6 ◽  
Author(s):  
Meng-Hsuan Ho ◽  
Zhong-Mao Guo ◽  
Julio Chunga ◽  
J. Shawn Goodwin ◽  
Hua Xie
Keyword(s):  
Endothelial Cells ◽  
Immune Responses ◽  
Outer Membrane ◽  
Porphyromonas Gingivalis ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Innate Immune ◽  
Innate Immune Responses ◽  
Human Endothelial Cells

An Investigation into the Effects of Outer Membrane Vesicles and Lipopolysaccharide of Porphyromonas gingivalis on Blood-Brain Barrier Integrity, Permeability, and Disruption of Scaffolding Proteins in a Human in vitro Model

2022 ◽  
pp. 1-22
Author(s):  
Anna Barlach Pritchard ◽  
Zsolt Fabian ◽  
Clare L. Lawrence ◽  
Glyn Morton ◽  
StJohn Crean ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Human Brain ◽  
Outer Membrane ◽  
Porphyromonas Gingivalis ◽  
Virulence Factors ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells

Background: The effects of the key pathogens and virulence factors associated with gum disease such as Porphyromonas gingivalis (P. gingivalis) on the central nervous system is of great interest with respect to development of neuropathologies and hence therapeutics and preventative strategies. Chronic infections and associated inflammation are known to weaken the first line of defense for the brain, the blood-brain barrier (BBB). Objective: The focus of this study is to utilize an established human in vitro BBB model to evaluate the effects of P. gingivalis virulence factors lipopolysaccharide (LPS) and outer membrane vesicles (OMVs) on a primary-derived human model representing the neurovascular unit of the BBB. Methods: Changes to the integrity of the BBB after application of P. gingivalis LPS and OMVs were investigated and correlated with transport of LPS. Additionally, the effect of P. gingivalis LPS and OMVs on human brain microvascular endothelial cells in monolayer was evaluated using immunofluorescence microscopy. Results: The integrity of the BBB model was weakened by application of P. gingivalis LPS and OMVs, as measured by a decrease in electrical resistance and a recovery deficit was seen in comparison to the controls. Application of P. gingivalis OMVs to a monoculture of human brain microvascular endothelial cells showed disruption of the tight junction zona occludens protein (ZO-1) compared to controls. Conclusion: These findings show that the integrity of tight junctions of the human BBB could be weakened by association with P. gingivalis virulence factors LPS and OMVs containing proteolytic enzymes (gingipains).

Outer membrane vesicles from pathogenic bacteria initiate an inflammatory response in human endothelial cells

2013 ◽  
Vol 184 (1) ◽  
pp. 458-466 ◽  
Author(s):  
Michael C. Soult ◽  
Natalie E. Lonergan ◽  
Bhairav Shah ◽  
Woong-Ki Kim ◽  
L.D. Britt ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Inflammatory Response ◽  
Outer Membrane ◽  
Pathogenic Bacteria ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Human Endothelial Cells

Electrophysiological Characterization of Transport Across Outer Membrane Channels from Gram-Negative Bacteria in Their Native Environment

2019 ◽  
Author(s):  
Jiajun Wang ◽  
Rémi Terrasse ◽  
Jayesh Arun Bafna ◽  
Lorraine Benier ◽  
Mathias Winterhalter
Keyword(s):  
Outer Membrane ◽  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Multi Drug Resistance ◽  
Gram Negative Bacteria ◽  
Membrane Channels ◽  
Gram Negative ◽  
Electrophysiological Characterization

Multi-drug resistance in Gram-negative bacteria is often associated with low permeability of the outer membrane. To investigate the role of membrane channels in the uptake of antibiotics, we extract, purify and reconstitute them into artificial planar membranes. To avoid this time-consuming procedure, here we show a robust approach using fusion of native outer membrane vesicles (OMV) into planar lipid bilayer which moreover allows also to some extend the characterization of membrane protein channels in their native environment. Two major membrane channels from <i>Escherichia coli</i>, OmpF and OmpC, were overexpressed from the host and the corresponding OMVs were collected. Each OMV fusion revealed surprisingly single or only few channel activities. The asymmetry of the OMV´s translates after fusion into the lipid membrane with the LPS dominantly present at the side of OMV addition. Compared to conventional reconstitution methods, the channels fused from OMVs containing LPS have similar conductance but a much broader distribution. The addition of Enrofloxacin on the LPS side yields somewhat higher association (<i>k<sub>on</sub></i>) and lower dissociation (<i>k<sub>off</sub></i>) rates compared to LPS-free reconstitution. We conclude that using outer membrane vesicles is a fast and easy approach for functional and structural studies of membrane channels in the native membrane.

A comprehensive characterization of membrane vesicles released by autophagic human endothelial cells

PROTEOMICS ◽  
2013 ◽  
Vol 13 (7) ◽  
pp. 1108-1120 ◽  
Author(s):  
Nicolas Pallet ◽  
Isabelle Sirois ◽  
Christina Bell ◽  
Laïla-Aïcha Hanafi ◽  
Katia Hamelin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Membrane Vesicles ◽  
Human Endothelial Cells ◽  
Comprehensive Characterization
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