Comparison of separation methods for immunomodulatory extracellular vesicles from helminths

Anne Borup; Anders T. Boysen; Andrea Ridolfi; Marco Brucale; Francesco Valle; Lucia Paolini; Paolo Bergese; Peter Nejsum

doi:10.1002/jex2.41

Comparison of separation methods for immunomodulatory extracellular vesicles from helminths

Anne Borup^*, Anders T. Boysen, Andrea Ridolfi, Marco Brucale, Francesco Valle, Lucia Paolini, Paolo Bergese, Peter Nejsum^*

^*Corresponding author af dette arbejde

Institut for Klinisk Medicin - Infektionssygdomme

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

Abstract

Abstract Helminths survive within their host by secreting immunomodulatory compounds, which hold therapeutic potential for inflammatory conditions. Helminth-derived extracellular vesicles (EVs) are one such component proposed to possess immunomodulatory activities. Due to the recent discovery of helminth EVs, standardised protocols for EV separation are lacking. Excretory/secretory products of the porcine helminth, Ascaris suum, were used to compare three EV separation methods: Size exclusion chromatography (SEC), ultracentrifugation (UC) and a combination of the two. Their performance was evaluated by EV yield, sample purity and the ability of EVs to suppress lipopolysaccharide (LPS)-induced inflammation in vitro. We found that all three separation methods successfully separated helminth EVs with a similar EV yield. Functional studies showed that EVs from all three methods reduced LPS-induced levels of tumour necrosis factor (TNF-α) in a dose-dependent manner. Overall, the three separation methods showed similar performance, however, the combination of UC+SEC presented with slightly higher purity than either method alone.

Originalsprog	Dansk
Artikelnummer	e41
Tidsskrift	Journal of Extracellular Biology
Vol/bind	1
Nummer	5
ISSN	2768-2811
DOI	https://doi.org/10.1002/jex2.41
Status	Udgivet - maj 2022

Emneord

Ascaris
extracellular vesicles
helminth
host-parasite interaction
immune modulation
separation methods

Adgang til dokumentet

10.1002/jex2.41Licens: CC BY

https://zenodo.org/record/6637514/files/J%20of%20Extracellular%20Bio%20-%202022%20-%20Borup%20-%20Comparison%20of%20separation%20methods%20for%20immunomodulatory%20extracellular%20vesicles%20from.pdfLicens: CC BY

Citationsformater

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title = "Comparison of separation methods for immunomodulatory extracellular vesicles from helminths",

abstract = "Abstract Helminths survive within their host by secreting immunomodulatory compounds, which hold therapeutic potential for inflammatory conditions. Helminth-derived extracellular vesicles (EVs) are one such component proposed to possess immunomodulatory activities. Due to the recent discovery of helminth EVs, standardised protocols for EV separation are lacking. Excretory/secretory products of the porcine helminth, Ascaris suum, were used to compare three EV separation methods: Size exclusion chromatography (SEC), ultracentrifugation (UC) and a combination of the two. Their performance was evaluated by EV yield, sample purity and the ability of EVs to suppress lipopolysaccharide (LPS)-induced inflammation in vitro. We found that all three separation methods successfully separated helminth EVs with a similar EV yield. Functional studies showed that EVs from all three methods reduced LPS-induced levels of tumour necrosis factor (TNF-α) in a dose-dependent manner. Overall, the three separation methods showed similar performance, however, the combination of UC+SEC presented with slightly higher purity than either method alone.",

keywords = "Ascaris, extracellular vesicles, helminth, host-parasite interaction, immune modulation, separation methods",

author = "Anne Borup and Boysen, {Anders T.} and Andrea Ridolfi and Marco Brucale and Francesco Valle and Lucia Paolini and Paolo Bergese and Peter Nejsum",

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T1 - Comparison of separation methods for immunomodulatory extracellular vesicles from helminths

AU - Borup, Anne

AU - Boysen, Anders T.

AU - Ridolfi, Andrea

AU - Brucale, Marco

AU - Valle, Francesco

AU - Paolini, Lucia

AU - Bergese, Paolo

AU - Nejsum, Peter

PY - 2022/5

Y1 - 2022/5

N2 - Abstract Helminths survive within their host by secreting immunomodulatory compounds, which hold therapeutic potential for inflammatory conditions. Helminth-derived extracellular vesicles (EVs) are one such component proposed to possess immunomodulatory activities. Due to the recent discovery of helminth EVs, standardised protocols for EV separation are lacking. Excretory/secretory products of the porcine helminth, Ascaris suum, were used to compare three EV separation methods: Size exclusion chromatography (SEC), ultracentrifugation (UC) and a combination of the two. Their performance was evaluated by EV yield, sample purity and the ability of EVs to suppress lipopolysaccharide (LPS)-induced inflammation in vitro. We found that all three separation methods successfully separated helminth EVs with a similar EV yield. Functional studies showed that EVs from all three methods reduced LPS-induced levels of tumour necrosis factor (TNF-α) in a dose-dependent manner. Overall, the three separation methods showed similar performance, however, the combination of UC+SEC presented with slightly higher purity than either method alone.

AB - Abstract Helminths survive within their host by secreting immunomodulatory compounds, which hold therapeutic potential for inflammatory conditions. Helminth-derived extracellular vesicles (EVs) are one such component proposed to possess immunomodulatory activities. Due to the recent discovery of helminth EVs, standardised protocols for EV separation are lacking. Excretory/secretory products of the porcine helminth, Ascaris suum, were used to compare three EV separation methods: Size exclusion chromatography (SEC), ultracentrifugation (UC) and a combination of the two. Their performance was evaluated by EV yield, sample purity and the ability of EVs to suppress lipopolysaccharide (LPS)-induced inflammation in vitro. We found that all three separation methods successfully separated helminth EVs with a similar EV yield. Functional studies showed that EVs from all three methods reduced LPS-induced levels of tumour necrosis factor (TNF-α) in a dose-dependent manner. Overall, the three separation methods showed similar performance, however, the combination of UC+SEC presented with slightly higher purity than either method alone.

KW - Ascaris

KW - extracellular vesicles

KW - helminth

KW - host-parasite interaction

KW - immune modulation

KW - separation methods

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DO - 10.1002/jex2.41

M3 - Tidsskriftartikel

SN - 2768-2811

VL - 1

JO - Journal of Extracellular Biology

JF - Journal of Extracellular Biology

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