Directed Protein Packaging within Outer Membrane Vesicles from &lt;em&gt;Escherichia coli&lt;/em&gt;: Design, Production and Purification

Alves, Nathan J.; Turner, Kendrick B.; Walper, Scott A.

doi:10.3791/54458

Cited by 12 publications

(11 citation statements)

References 30 publications

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“…On the basis of the previous studies (Alves, Turner, & Walper, 2016;Ho, Wu, Chang, & Pan, 2011;Pollak, Delpino, Fossati, & Baldi, 2012;Rider, Hurwitz, & Meckes, 2016;Thery, Amigorena, Raposo, & Clayton, 2006;Wieser et al, 2014), the OMVs extraction and purification methods were as follows:…”

Section: Extraction and Purification Of Omvs In Faecesmentioning

confidence: 99%

Outer membrane vesicles enhance tau phosphorylation and contribute to cognitive impairment

Wei

Peng

Mai

et al. 2019

Journal Cellular Physiology

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Outer membrane vesicles (OMVs) are nanosized vesicles produced by the gut microbiota (GM). The GM is well-known to be involved in the pathological process of Alzheimer's disease (AD). However, the mechanism of OMVs is not clear. In the present study, we demonstrated the involvement of OMVs in the development of cognitive (learning and memory) dysfunction induced by blood-brain barrier (BBB) disruption. More important, further study showed that OMVs induced tau phosphorylation by activating glycogen synthase kinase 3β (GSK-3β) in the hippocampus. OMVs activated astrocytes and microglia, increased secretion of inflammatory cytokines (nuclear factor κB, interleukin-1β, and tumour necrosis factor-α) in the hippocampus. Therefore, OMVs increase the permeability of the BBB and promote the activation of astrocytes and microglia, inducing an inflammatory response and tau hyperphosphorylation by activating the GSK-3β pathway and finally leading to cognitive impairment. K E Y W O R D SAlzheimer's disease, astrocytes, blood-brain barrier, glycogen synthase kinase 3β, gut microbiota, inflammatory cytokines, interleukin-1β, microglia, nuclear factor κB, outer membrane vesicles, tau, tumour necrosis factor-α

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Section: Extraction and Purification Of Omvs In Faecesmentioning

confidence: 99%

Outer membrane vesicles enhance tau phosphorylation and contribute to cognitive impairment

Wei

Peng

Mai

et al. 2019

Journal Cellular Physiology

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“…MVs protect their encased cargo from degradation by nucleases [ 29 , 65 ] and proteases [ 66 , 67 , 68 , 69 ]. MVs have been shown to retain enzymatic activity and antigenicity over long-term storage at 4 °C [ 70 , 71 ], as well as short exposure to elevated temperatures. A study has shown that an enzyme, phosphotriesterase (PTE) packaged into E. coli MVs retained enzymatic activity over 100-fold greater than free PTE when subject to elevated temperatures (37 °C for 14 days), and multiple freeze-thaw cycles [ 66 ].…”

Section: Advantages Of Mvs In Therapeutic Applicationsmentioning

confidence: 99%

“…Genetic manipulation of the parent bacteria can refine the functionality of MVs, for instance it allows directed packaging of recombinant epitopes, inclusion of signal molecules for cell-specific targeting or exclusion of undesired and toxic MV components such as LPS [ 56 , 76 , 107 ]. This relative ease of biomanipulation is leading to the development of unique recombinant MV therapies [ 70 ].…”

Section: Bioengineered Mvsmentioning

confidence: 99%

“…This approach was used to encapsulate functional green fluorescence protein (GFP) into E. coli MVs using the Tat signal sequence [ 111 ]. Surface display of recombinant proteins, which is required for the display of many surface antigens, may be achieved through fusion to a surface anchor protein, such as an abundant outer membrane protein or glycoprotein [ 70 , 112 ] ( Figure 2 c). This results in localization of the antigen to the MV surface, where it can be efficiently presented to the immune system [ 113 ].…”

Section: Bioengineered Mvsmentioning

confidence: 99%

“…Furthermore, careful selection of the anchor protein is required to ensure effective incorporation and display of the recombinant epitope without disrupting MV formation or growth of the parent bacteria [ 70 ]. The first study to prove this concept demonstrated that several heterologous proteins could be displayed on the surface of E. coli and Salmonella enterica MVs by genetically fusing them to the surface protein cytolysin A (ClyA) [ 114 ].…”

Section: Bioengineered Mvsmentioning

confidence: 99%

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The Therapeutic Benefit of Bacterial Membrane Vesicles

Bitto

Kaparakis‐Liaskos

2017

IJMS

116

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The therapeutic potential of extracellular vesicles from eukaryotes has gained strong interest in recent years. However, research into the therapeutic application of their bacterial counterparts, known as bacterial membrane vesicles, is only just beginning to be appreciated. Membrane vesicles (MVs) from both Gram-positive and Gram-negative bacteria offer significant advantages in therapeutic development, including large-scale, cost effective production and ease of molecular manipulation to display foreign antigens. The nanoparticle size of MVs enables their dissemination through numerous tissue types, and their natural immunogenicity and self-adjuvanting capability can be harnessed to induce both cell-mediated and humoral immunity in vaccine design. Moreover, the ability to target MVs to specific tissues through the display of surface receptors raises their potential use as targeted MV-based anti-cancer therapy. This review discusses recent advances in MV research with particular emphasis on exciting new possibilities for the application of MVs in therapeutic design.

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Isolation of Outer Membrane Vesicles Including Their Quantitative and Qualitative Analyses

Kohl

Zingl

Eichmann

et al. 2018

Methods in Molecular Biology

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Outer membrane vesicles (OMVs) are naturally secreted from the bacterial cell surface and therefore localized in the cell-free supernatant of bacterial cultures. Here we describe methods for crude and density gradient-purified OMV isolation and protocols for control analyses for protein profiling (SDS-PAGE), detection of indicator proteins (immunoblot analysis), lipid profiling (lipid extraction and LC-MS analysis), vesicle size determination (NanoSight), rough estimation of biomass (TrayCell™), as well as quantifications of defined OMV components, e.g., proteins (Bradford) and LPS (Purpald).

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Directed Protein Packaging within Outer Membrane Vesicles from <em>Escherichia coli</em>: Design, Production and Purification

Cited by 12 publications

References 30 publications

Outer membrane vesicles enhance tau phosphorylation and contribute to cognitive impairment

Outer membrane vesicles enhance tau phosphorylation and contribute to cognitive impairment

The Therapeutic Benefit of Bacterial Membrane Vesicles

Isolation of Outer Membrane Vesicles Including Their Quantitative and Qualitative Analyses

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