Enterotoxigenic Escherichia coli vesicles target toxin delivery into mammalian cells

Kesty, Nicole C.; Mason, Kevin M.; Reedy, Mary C.; Miller, Sara; Kuehn, Meta J.

doi:10.1038/sj.emboj.7600471

Cited by 324 publications

(308 citation statements)

References 60 publications

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“…Internalization of OMVs in human cells, which may also allow further intracellular trafficking of the vesicles, is in accordance with several recent studies on OMVs from various bacterial species, including the human pathogens B. abortus, E. coli, H. pylori, P. aeruginosa, P. gingivalis, and V. cholerae (23)(24)(25)(26)(27)(28) and also nonvirulent E. coli K-12 strains (63). Colocalization of A. actinomycetemcomitans OMVs with the ER is consistent with findings with vesicles from enterotoxigenic E. coli and P. aeruginosa (24,26). However, similar to observations with the P. aeruginosa OMVs (50), intracellular delivery of A. actinomycetemcomitans vesicle cargo was unaffected by inhibition of retrograde transport (7), suggesting that passage through this pathway is not crucial for the antigen delivery.…”

Section: Discussionsupporting

confidence: 87%

“…It was not known if A. actinomycetemcomitans OMVs can be internalized into the interior of nonphagocytic human cells via endocytic pathways, analogously to vesicles derived from other species, such as Brucella abortus, Escherichia coli, Helicobacter pylori, Pseudomonas aeruginosa, Porphyromonas gingivalis, and Vibrio cholerae (23)(24)(25)(26)(27)(28). In addition to protein delivery, internalization of OMVs represents an important mechanism to expose the host cells to pathogen-associated molecular patterns (PAMPs), e.g., peptidoglycan (PGN) fragments and LPS, which are associated with intact OMVs and/or with fragments from ruptured vesicles within the cell and are recognized by intracellular host pattern recognition receptors (PRRs).…”

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confidence: 99%

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Aggregatibacter actinomycetemcomitans Outer Membrane Vesicles Are Internalized in Human Host Cells and Trigger NOD1- and NOD2-Dependent NF-κB Activation

et al. 2014

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Section: Discussionsupporting

confidence: 87%

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confidence: 99%

Aggregatibacter actinomycetemcomitans Outer Membrane Vesicles Are Internalized in Human Host Cells and Trigger NOD1- and NOD2-Dependent NF-κB Activation

et al. 2014

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“…Outer membrane vesicles (OMVs) were harvested from culture supernatants of bacteria grown overnight in 50 ml LB at 37°C with shaking (150 rpm) according to the method of Kesty et al (21). Bacteria were pelleted by centrifugation (10,000 ϫ g, 10 min, 4°C), and the supernatant was decanted and passed through a 0.2-m filter.…”

Section: Methodsmentioning

confidence: 99%

EnterotoxigenicEscherichia coliModulates Host Intestinal Cell Membrane Asymmetry and Metabolic Activity

et al. 2009

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Enterotoxigenic Escherichia coli (ETEC) is a common cause of travelers' and postweaning diarrhea in humans and swine, respectively. The extent to which ETEC damages host cells is unclear. Experiments are presented that probe the ability of porcine ETEC isolates to induce apoptosis and cell death in porcine intestinal epithelial cells. Quantification of host phosphatidylserine exposure following ETEC infection suggested that ETEC induced changes in plasma membrane asymmetry, independent of the expression of the heat-labile enterotoxin. Significant host cell death was not observed. ETEC infection also caused a drastic inhibition of host esterase activity, as measured by calcein fluorescence. While ETEC infection resulted in activation of host caspase 3, terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling of DNA double-strand breakage, indicative of late stages of apoptosis, was not observed. Camptothecin-induced apoptosis markedly increased subsequent ETEC adherence. Transfer of cell-free supernatants from apoptotic cells to bacterial inocula prior to infection of naïve cells increased the transcriptional activity of the regulatory region upstream of the K88ac operon and promoted subsequent adherence to host cells.Enterotoxigenic Escherichia coli (ETEC) causes childhood mortality and travelers' diarrhea in humans (36) and postweaning diarrhea in pigs (32). ETEC adherence to intestinal cells is mediated by proteinaceous, species-specific colonization factors (25). ETEC induces water and electrolyte loss from the intestine due to the activities of several enterotoxins, including the heat-labile enterotoxin (LT), the heat-stable toxins (ST), and the enteroaggregative E. coli heat-stable enterotoxin 1 (32).Bacterial pathogens often induce host cell apoptosis to promote their survival and dissemination (16,30). While several E. coli serotypes (1, 4, 9, 15) and toxins (3, 19) have been shown to induce apoptosis of epithelial cells, the extent to which ETEC damages host cells is unclear. The prototypic ETEC isolate H10407 was cytotoxic to but failed to induce apoptosis in the macrophage cell line J774 (as measured by DNA fragmentation) (24). The extent to which LT induces apoptosis has also received considerable attention due to interest in the receptor-binding subunit (LT-B) as a mucosal adjuvant. LT-B induces apoptosis of CD8 ϩ but not CD4 ϩ T cells (37). Some studies indicate that binding of LT-B to GM1 may be sufficient for triggering apoptosis of CD8 ϩ T cells (33), while others suggest that modified LT-B constructs (e.g., H57S) still able to bind GM-1 may fail to trigger apoptosis (14). Yet other studies suggest that LT-mediated apoptosis requires the ADP-ribosylation activity of the LT-A subunit (41).Overall, the roles of ETEC and LT in mediating apoptosis have not been defined clearly and remain somewhat controversial. The goals of this study were therefore to quantify the ability of ETEC to damage porcine intestinal epithelial cells, to clarify the role of LT in these processes, and to me...

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“…Incubation of purified P. aeruginosa OMV with eukaryotic cells lead to the fusion of the OMV with lipid rafts present in eukaryotic membranes, with the concomitant release of multiple virulence factors into the host cytosol (11). In other cases, whole OMV were internalized and incorporated into the trafficking network of the host cells (9,12). Based on these observations, it has been proposed that OMV act as long distance toxin delivery devices (4,11).…”

mentioning

confidence: 99%

Selective Sorting of Cargo Proteins into Bacterial Membrane Vesicles

Haurat¹,

Aduse‐Opoku

Rangarajan

et al. 2011

Journal of Biological Chemistry

272

290

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In contrast to the well established multiple cellular roles of membrane vesicles in eukaryotic cell biology, outer membrane vesicles (OMV) produced via blebbing of prokaryotic membranes have frequently been regarded as cell debris or microscopy artifacts. Increasingly, however, bacterial membrane vesicles are thought to play a role in microbial virulence, although it remains to be determined whether OMV result from a directed process or from passive disintegration of the outer membrane. Here we establish that the human oral pathogen Porphyromonas gingivalis has a mechanism to selectively sort proteins into OMV, resulting in the preferential packaging of virulence factors into OMV and the exclusion of abundant outer membrane proteins from the protein cargo. Furthermore, we show a critical role for lipopolysaccharide in directing this sorting mechanism. The existence of a process to package specific virulence factors into OMV may significantly alter our current understanding of host-pathogen interactions.

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Enterotoxigenic Escherichia coli vesicles target toxin delivery into mammalian cells

Cited by 324 publications

References 60 publications

Aggregatibacter actinomycetemcomitans Outer Membrane Vesicles Are Internalized in Human Host Cells and Trigger NOD1- and NOD2-Dependent NF-κB Activation

Aggregatibacter actinomycetemcomitans Outer Membrane Vesicles Are Internalized in Human Host Cells and Trigger NOD1- and NOD2-Dependent NF-κB Activation

EnterotoxigenicEscherichia coliModulates Host Intestinal Cell Membrane Asymmetry and Metabolic Activity

Selective Sorting of Cargo Proteins into Bacterial Membrane Vesicles

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