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Helicobacter pylori ATCC 43629/NCTC 11639 Outer Membrane Vesicles (OMVs) from biofilm and planktonic phase associated with extracellular DNA (eDNA)

Helicobacter pylori ATCC 43629/NCTC 11639 Outer Membrane Vesicles (OMVs) from biofilm and planktonic phase associated with extracellular DNA (eDNA)
Helicobacter pylori ATCC 43629/NCTC 11639 Outer Membrane Vesicles (OMVs) from biofilm and planktonic phase associated with extracellular DNA (eDNA)
Helicobacter pylori persistence is associated with its capacity to develop biofilms as a response to changing environmental conditions and stress. Extracellular DNA (eDNA) is a component of H. pylori biofilm matrix but the lack of DNase I activity supports the hypothesis that eDNA might be protected by other extracellular polymeric substances (EPS) and/or Outer Membrane Vesicles (OMVs), which bleb from the bacteria surface during growth. The aim of the present study was to both identify the eDNA presence on OMVs segregated from H. pylori ATCC 43629/NCTC 11639 biofilm (bOMVs) and its planktonic phase (pOMVs) and to characterize the physical-chemical properties of the OMVs. The presence of eDNA in bOMVs and pOMVs was initially carried out using DNase I-gold complex labeling and Transmission Electron Microscope analysis (TEM). bOMVs and pOMVs were further isolated and physical-chemical characterization carried out using dynamic light scattering (DLS) analysis. eDNA associated with OMVs was detected and quantified using a PicoGreen spectrophotometer assay, while its extraction was performed with a DNA Kit. TEM images showed that eDNA was mainly associated with the OMV membrane surfaces; while PicoGreen staining showed a four-fold increase of dsDNA in bOMVs compared with pOMVs. The eDNA extracted from OMVs was visualized using gel electrophoresis. DLS analysis indicated that both planktonic and biofilm H. pylori phenotypes generated vesicles, with a broad distribution of sizes on the nanometer scale. The DLS aggregation assay suggested that eDNA may play a role in the aggregation of OMVs, in the biofilm phenotype. Moreover, the eDNA associated with vesicle membrane may impede DNase I activity on H. pylori biofilms. These results suggest that OMVs derived from the H. pylori biofilm phenotype may play a structural role by preventing eDNA degradation by nucleases, providing a bridging function between eDNA strands on OMV surfaces and promoting aggregation.
helicobacter pylori, eDNA, outer membrane vesicles (OMVs), biofilm, nanoparticles
1664-302X
1-11
Grande, Rossella
acd0039e-d03a-498d-a1a4-c63db5a57d59
Di Marcantonio, Maria C.
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Robuffo, Iole
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Pompilio, Arianna
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Celia, Christian
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Di Marzio, Luisa
6b9be2b3-59a5-4c54-b85e-78d507d9e002
Paolino, Donatella
087ab0bb-d673-4c79-86ea-f465bc8ee769
Codagnone, Marilina
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Muraro, Raffaella
eb18adb8-10ed-4256-a5a8-fb5085497fdd
Stoodley, Paul
08614665-92a9-4466-806e-20c6daeb483f
Hall-Stoodley, Luanne
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Mincione, Gabriella
25deb07b-f154-4658-9f08-1346d70b5f84
Grande, Rossella
acd0039e-d03a-498d-a1a4-c63db5a57d59
Di Marcantonio, Maria C.
9c589f51-adfc-4666-b198-cea30b372865
Robuffo, Iole
be5ff069-9aa5-4cec-bac4-8e5530f75584
Pompilio, Arianna
f543fe14-78d3-4d05-9761-f02119f4f783
Celia, Christian
a83c1085-b231-45eb-9ac2-2f2baabc27ed
Di Marzio, Luisa
6b9be2b3-59a5-4c54-b85e-78d507d9e002
Paolino, Donatella
087ab0bb-d673-4c79-86ea-f465bc8ee769
Codagnone, Marilina
12dc1176-6e42-46c0-96d3-54d981e4ab71
Muraro, Raffaella
eb18adb8-10ed-4256-a5a8-fb5085497fdd
Stoodley, Paul
08614665-92a9-4466-806e-20c6daeb483f
Hall-Stoodley, Luanne
94ebdc00-b549-4488-b15f-5310fb965f5b
Mincione, Gabriella
25deb07b-f154-4658-9f08-1346d70b5f84

Grande, Rossella, Di Marcantonio, Maria C., Robuffo, Iole, Pompilio, Arianna, Celia, Christian, Di Marzio, Luisa, Paolino, Donatella, Codagnone, Marilina, Muraro, Raffaella, Stoodley, Paul, Hall-Stoodley, Luanne and Mincione, Gabriella (2015) Helicobacter pylori ATCC 43629/NCTC 11639 Outer Membrane Vesicles (OMVs) from biofilm and planktonic phase associated with extracellular DNA (eDNA). Frontiers in Microbiology, 6, 1-11. (doi:10.3389/fmicb.2015.01369). (PMID:26733944)

Record type: Article

Abstract

Helicobacter pylori persistence is associated with its capacity to develop biofilms as a response to changing environmental conditions and stress. Extracellular DNA (eDNA) is a component of H. pylori biofilm matrix but the lack of DNase I activity supports the hypothesis that eDNA might be protected by other extracellular polymeric substances (EPS) and/or Outer Membrane Vesicles (OMVs), which bleb from the bacteria surface during growth. The aim of the present study was to both identify the eDNA presence on OMVs segregated from H. pylori ATCC 43629/NCTC 11639 biofilm (bOMVs) and its planktonic phase (pOMVs) and to characterize the physical-chemical properties of the OMVs. The presence of eDNA in bOMVs and pOMVs was initially carried out using DNase I-gold complex labeling and Transmission Electron Microscope analysis (TEM). bOMVs and pOMVs were further isolated and physical-chemical characterization carried out using dynamic light scattering (DLS) analysis. eDNA associated with OMVs was detected and quantified using a PicoGreen spectrophotometer assay, while its extraction was performed with a DNA Kit. TEM images showed that eDNA was mainly associated with the OMV membrane surfaces; while PicoGreen staining showed a four-fold increase of dsDNA in bOMVs compared with pOMVs. The eDNA extracted from OMVs was visualized using gel electrophoresis. DLS analysis indicated that both planktonic and biofilm H. pylori phenotypes generated vesicles, with a broad distribution of sizes on the nanometer scale. The DLS aggregation assay suggested that eDNA may play a role in the aggregation of OMVs, in the biofilm phenotype. Moreover, the eDNA associated with vesicle membrane may impede DNase I activity on H. pylori biofilms. These results suggest that OMVs derived from the H. pylori biofilm phenotype may play a structural role by preventing eDNA degradation by nucleases, providing a bridging function between eDNA strands on OMV surfaces and promoting aggregation.

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e-pub ahead of print date: 17 November 2015
Published date: 16 December 2015
Keywords: helicobacter pylori, eDNA, outer membrane vesicles (OMVs), biofilm, nanoparticles
Organisations: Bioengineering Group, Engineering Science Unit, nCATS Group, Faculty of Engineering and the Environment

Identifiers

Local EPrints ID: 385967
URI: http://eprints.soton.ac.uk/id/eprint/385967
ISSN: 1664-302X
PURE UUID: fa2f47ec-bcba-4566-badc-25e992a6c1f8
ORCID for Paul Stoodley: ORCID iD orcid.org/0000-0001-6069-273X

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Date deposited: 27 Jan 2016 09:24
Last modified: 26 Nov 2019 01:41

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Contributors

Author: Rossella Grande
Author: Maria C. Di Marcantonio
Author: Iole Robuffo
Author: Arianna Pompilio
Author: Christian Celia
Author: Luisa Di Marzio
Author: Donatella Paolino
Author: Marilina Codagnone
Author: Raffaella Muraro
Author: Paul Stoodley ORCID iD
Author: Luanne Hall-Stoodley
Author: Gabriella Mincione

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