Heat shock enhances outer-membrane vesicle release in Bordetella spp.

Jonge, Eline F. de; Balhuizen, Melanie D.; Boxtel, Ria van; Wu, Jianjun; Haagsman, Henk P.; Tommassen, Jan

doi:10.1016/j.crmicr.2020.100009

Cited by 22 publications

(32 citation statements)

References 66 publications

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“…However, analysis of the LPS of the heat-killed cells that were used in the TLR4 activation assays by SDS-PAGE indicated that the LPS in the wild-type samples is fully palmitoylated (Figure S1), suggesting that PagP is activated during the heat treatment. This hypothesis is in line with our previous studies in which we used the same heat treatment to stimulate the release of outer-membrane vesicles (OMVs) [29]. Lipidomic analysis showed a large increase in lysophospholipid content in heat-released OMVs compared to spontaneously released OMVs [43].…”

Section: Discussionsupporting

confidence: 91%

“…Alternatively, the separated proteins were transferred to a 0.45-μm pore-size nitrocellulose membrane (GE Healthcare). For immunodetection, mouse antiserum directed against autotransporter BrkA, generously provided by Nathalie Devos (GlaxoSmithKline Biologicals SA) and rabbit antisera directed against the major porin OmpP and siderophore receptor FauA [29] were used. As secondary antibodies, horseradish peroxidase-conjugated goat anti-mouse or antirabbit IgG antisera (ThermoFisher) were employed.…”

Section: Outer Membrane Isolation and Analysismentioning

confidence: 99%

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Reduction of endotoxicity in Bordetella bronchiseptica by lipid A engineering: Characterization of lpxL1 and pagP mutants

et al. 2021

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Whole-cell vaccines against Gram-negative bacteria commonly display high reactogenicity caused by the endotoxic activity of lipopolysaccharide (LPS), one of the major components of the bacterial outer membrane. Underacylation of the lipid A moiety of LPS has been related with reduced endotoxicity in several Gram-negative species. Here, we evaluated whether the inactivation of two genes encoding lipid A acylases of Bordetella bronchiseptica, i.e. pagP and lpxL1, could be used for the development of less reactogenic vaccines against this pathogen for livestock and companion animals. Inactivation of pagP resulted in the loss of the secondary palmitate chain at position 3' of lipid A, but hardly affected the potency of the LPS to activate the Toll-like receptor 4 (TLR4). Inactivation of lpxL1 resulted in the loss of the secondary 2-hydroxy laurate group present at position 2 of lipid A and, unexpectedly, in the additional loss of the glucosamines that decorate the phosphate groups at positions 1 and 4' and in an increase in LPS molecules carrying O-antigen. The resulting LPS showed greatly reduced potency to activate TLR4 in HEK-Blue reporter cells expressing human or mouse TLR4 as well as in porcine macrophages. Characterization of the lpxL1 mutant revealed many pleiotropic phenotypes, including increased resistance to SDS and rifampicin, increased susceptibility to cationic antimicrobial peptides, decreased auto-aggregation and biofilm formation, and a tendency to decreased infectivity of macrophages, which are all related to the altered LPS structure. We suggest that the lpxL1 mutant will be useful for the generation of safer vaccines.

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Section: Discussionsupporting

confidence: 91%

Section: Outer Membrane Isolation and Analysismentioning

confidence: 99%

Reduction of endotoxicity in Bordetella bronchiseptica by lipid A engineering: Characterization of lpxL1 and pagP mutants

et al. 2021

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“…Bacterial cultures were stimulated with two sublethal concentrations of different peptides, PMAP-36, CATH-2, and LL-37, and peptide-induced OMVs (pOMVs) were isolated. Heat treatment was applied as a control stressor, since it has been shown to induce OMV release, resulting in heat-induced OMVs (hOMVs) ( 22 , 23 ). Isolated pOMVs and hOMVs were analyzed using Coomassie-stained SDS-PAGE and compared to spontaneous OMVs (sOMVs; Fig.…”

Section: Resultsmentioning

confidence: 99%

Outer Membrane Vesicles Protect Gram-Negative Bacteria against Host Defense Peptides

Balhuizen

Dijk

Jansen

et al. 2021

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“…Similarly, after treatment with higher temperatures B-band LPS export in OMVs was increased in P. aeruginosa ( Eberlein et al, 2018 ). Recently it was shown that heat treatment also increased OMV production in B. pertussis ( de Jonge et al, 2021 ). These heat-induced OMVs (hOMVs) were shown to still contain important antigens, which could be detected with antibodies.…”

Section: Future Prospectsmentioning

confidence: 99%

Outer Membrane Vesicle Induction and Isolation for Vaccine Development

2021

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Gram-negative bacteria release vesicular structures from their outer membrane, so called outer membrane vesicles (OMVs). OMVs have a variety of functions such as waste disposal, communication, and antigen or toxin delivery. These vesicles are the promising structures for vaccine development since OMVs carry many surface antigens that are identical to the bacterial surface. However, isolation is often difficult and results in low yields. Several methods to enhance OMV yield exist, but these do affect the resulting OMVs. In this review, our current knowledge about OMVs will be presented. Different methods to induce OMVs will be reviewed and their advantages and disadvantages will be discussed. The effects of the induction and isolation methods used in several immunological studies on OMVs will be compared. Finally, the challenges for OMV-based vaccine development will be examined and one example of a successful OMV-based vaccine will be presented.

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Heat shock enhances outer-membrane vesicle release in Bordetella spp.

Cited by 22 publications

References 66 publications

Reduction of endotoxicity in Bordetella bronchiseptica by lipid A engineering: Characterization of lpxL1 and pagP mutants

Reduction of endotoxicity in Bordetella bronchiseptica by lipid A engineering: Characterization of lpxL1 and pagP mutants

Outer Membrane Vesicles Protect Gram-Negative Bacteria against Host Defense Peptides

Outer Membrane Vesicle Induction and Isolation for Vaccine Development

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