Lipopolysaccharide structure impacts the entry kinetics of bacterial outer membrane vesicles into host cells

Eloise J O'Donoghue; Natalie Sirisaengtaksin; Douglas F Browning; Ewa Bielska; Mohammed Hadis; Francisco Fernandez-Trillo; Luke Alderwick; Sara Jabbari; Anne Marie Krachler

doi:10.1371/journal.ppat.1006760

Lipopolysaccharide structure impacts the entry kinetics of bacterial outer membrane vesicles into host cells

Eloise J O'Donoghue, Natalie Sirisaengtaksin, Douglas F Browning, Ewa Bielska, Mohammed Hadis, Francisco Fernandez-Trillo, Luke Alderwick, Sara Jabbari, Anne Marie Krachler

Research output: Contribution to journal › Article › peer-review

Abstract

Outer membrane vesicles are nano-sized microvesicles shed from the outer membrane of Gram-negative bacteria and play important roles in immune priming and disease pathogenesis. However, our current mechanistic understanding of vesicle-host cell interactions is limited by a lack of methods to study the rapid kinetics of vesicle entry and cargo delivery to host cells. Here, we describe a highly sensitive method to study the kinetics of vesicle entry into host cells in real-time using a genetically encoded, vesicle-targeted probe. We found that the route of vesicular uptake, and thus entry kinetics and efficiency, are shaped by bacterial cell wall composition. The presence of lipopolysaccharide O antigen enables vesicles to bypass clathrin-mediated endocytosis, which enhances both their entry rate and efficiency into host cells. Collectively, our findings highlight the composition of the bacterial cell wall as a major determinant of secretion-independent delivery of virulence factors during Gram-negative infections.

Original language	English
Article number	e1006760
Journal	Plos Pathogens
Volume	13
Issue number	11
DOIs	https://doi.org/10.1371/journal.ppat.1006760
Publication status	Published - 29 Nov 2017

Bibliographical note

© 2017 O’Donoghue et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: This work was supported by Biotechnology and Biological Sciences Research Council (http://www.bbsrc.ac.uk/) grants BB/M021513/1 and BB/L007916/1 (to AMK), and a BBSRC MIBTP studentship (to EJO).

Keywords

Cell Wall/chemistry
Endocytosis
Gram-Negative Bacteria/chemistry
Gram-Negative Bacterial Infections/metabolism
Host-Pathogen Interactions
Humans
Kinetics
Lipopolysaccharides/chemistry
Transport Vesicles/metabolism
Virulence Factors/metabolism

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10.1371/journal.ppat.1006760Licence: CC BY 3.0

Lipopolysaccharide structure impacts the entry kinetics
© 2017 O’Donoghue et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Final published version, 8.6 MBLicence: CC BY 3.0

Cite this

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abstract = "Outer membrane vesicles are nano-sized microvesicles shed from the outer membrane of Gram-negative bacteria and play important roles in immune priming and disease pathogenesis. However, our current mechanistic understanding of vesicle-host cell interactions is limited by a lack of methods to study the rapid kinetics of vesicle entry and cargo delivery to host cells. Here, we describe a highly sensitive method to study the kinetics of vesicle entry into host cells in real-time using a genetically encoded, vesicle-targeted probe. We found that the route of vesicular uptake, and thus entry kinetics and efficiency, are shaped by bacterial cell wall composition. The presence of lipopolysaccharide O antigen enables vesicles to bypass clathrin-mediated endocytosis, which enhances both their entry rate and efficiency into host cells. Collectively, our findings highlight the composition of the bacterial cell wall as a major determinant of secretion-independent delivery of virulence factors during Gram-negative infections.",

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KW - Virulence Factors/metabolism

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Lipopolysaccharide structure impacts the entry kinetics of bacterial outer membrane vesicles into host cells

Abstract

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Keywords

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