scholarly journals Hydrostatic Filtration Enables Large-Scale Production of Outer Membrane Vesicles That Effectively Protect Chickens against Gallibacterium anatis

Vaccines ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 40
Author(s):  
Fabio Antenucci ◽  
Homa Arak ◽  
Jianyang Gao ◽  
Toloe Allahgadry ◽  
Ida Thøfner ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Egg Production ◽  
Large Scale ◽  
Membrane Vesicles ◽  
Optimal Dose ◽  
Outer Membrane Vesicles ◽  
Poultry Meat ◽  
Scale Production ◽  
Layer Chickens ◽  
N Proteins

Gallibacterium anatis is a Gram-negative opportunistic avian pathogen representing an emerging threat to poultry meat and egg production worldwide. To date, no vaccine able to effectively prevent the morbidity associated with G. anatis infections has been developed yet. Our group previously reported that inoculation of different combinations of G. anatis outer membrane vesicles (OMVs), FlfA and GtxA-N proteins is effective in preventing lesions caused by G. anatis infections in layer chickens. Here we report the testing of the efficacy as vaccine prototypes of G. anatis OMVs isolated by hydrostatic filtration, a simple technique that allows the cost-effective isolation of high yields of OMVs. Layer chickens were immunized with OMVs alone or in combination with FlfA and/or GtxA-N proteins. Subsequent challenge with a heterologous G. anatis strain showed that immunization with OMVs alone could significantly reduce the lesions following a G. anatis infection. A second study was carried out to characterize the dose-response (0.25, 2.5 and 25 µg) relationship of G. anatis OMVs as immunogens, showing that 2.5 μg of OMVs represent the optimal dose to elicit protection in the immunized animals after a similar challenge. Additionally, administration of ≥2.5 μg of G. anatis OMVs induced specific IgY titers and possibly vertical transfer of immunity.

​Optimization of Cultural Conditions for Maximum Production of Outer Membrane Vesicles from Salmonella Typhimurium

2021 ◽  
Author(s):  
Menguzotunuo Solo ◽  
Shantanu Tamuly ◽  
Luit Moni Barkalita ◽  
Girin Kumar Saikia ◽  
Dhruba Jyoti Kalita
Keyword(s):  
Oxidative Stress ◽  
Salmonella Typhimurium ◽  
Outer Membrane ◽  
Large Scale ◽  
Multiple Drug Resistance ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Multiple Drug ◽  
Scale Production ◽  
Maximum Production

Background: The non-typhoidal Salmonella causes gastroenteritis in humans that makes its way to the food chain mainly through the animal products. The multiple drug resistance imposes one of the major hurdle in the treatment of the disease. The vaccination appears to be the most important method for prevention of the disease. Unfortunately, there is no liscenced vaccine available against non-typhoidal Salmonellae. The use of outer membrane vesicles (OMVs) of Salmonella as a vaccine candidate has attained significant centre-stage in the recent years given to its protective immunogenicity. However, the large scale production of OMVs is difficult owing to low yield per liter of culture. Methods: In the present study, we have optimized the culture conditions viz. pH, phase of growth and presence of oxidative stress for maximum production of OMVs from Salmonella Typhimurium. The OMVs were characterized based on yield based on protein concentration, lipopolysaccharide concentration and zeta size. Result: In the present study, it was found that incubation of Salmonella Typhimurium up to peak of the growth phase at pH 7 in presence of oxidative stress was found to be the most suitable condition for maximum production of OMVs.

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.

Outer Membrane Vesicles Facilitate Trafficking of the Hydrophobic Signalling Molecule CAI-1 Between Vibrio harveyi Cells

2018 ◽  
Author(s):  
Sophie Brameyer ◽  
Laure Plener ◽  
Axel MMller ◽  
Andreas Klingl ◽  
Gerhard Wanner ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Vibrio Harveyi ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Signalling Molecule ◽  
I Cells

Preparation of Outer Membrane Vesicles from Myxococcus xanthus

BIO-PROTOCOL ◽  
2016 ◽  
Vol 6 (2) ◽  
Author(s):  
James Berleman ◽  
Marcin Zemla ◽  
Jonathan Remis ◽  
Manfred Auer
Keyword(s):  
Outer Membrane ◽  
Myxococcus Xanthus ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles

scholarly journals Helicobacter pylori Outer Membrane Vesicles and Extracellular Vesicles from Helicobacter pylori-Infected Cells in Gastric Disease Development

2021 ◽  
Vol 22 (9) ◽  
pp. 4823
Author(s):  
María Fernanda González ◽  
Paula Díaz ◽  
Alejandra Sandoval-Bórquez ◽  
Daniela Herrera ◽  
Andrew F. G. Quest
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Outer Membrane ◽  
Extracellular Vesicles ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Host Cells ◽  
Gastric Epithelium ◽  
Infected Cells ◽  
H Pylori

Extracellular vesicles (EVs) are cell-derived vesicles important in intercellular communication that play an essential role in host-pathogen interactions, spreading pathogen-derived as well as host-derived molecules during infection. Pathogens can induce changes in the composition of EVs derived from the infected cells and use them to manipulate their microenvironment and, for instance, modulate innate and adaptive inflammatory immune responses, both in a stimulatory or suppressive manner. Gastric cancer is one of the leading causes of cancer-related deaths worldwide and infection with Helicobacter pylori (H. pylori) is considered the main risk factor for developing this disease, which is characterized by a strong inflammatory component. EVs released by host cells infected with H. pylori contribute significantly to inflammation, and in doing so promote the development of disease. Additionally, H. pylori liberates vesicles, called outer membrane vesicles (H. pylori-OMVs), which contribute to atrophia and cell transformation in the gastric epithelium. In this review, the participation of both EVs from cells infected with H. pylori and H. pylori-OMVs associated with the development of gastric cancer will be discussed. By deciphering which functions of these external vesicles during H. pylori infection benefit the host or the pathogen, novel treatment strategies may become available to prevent disease.

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