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Fluorescent labeling of helminth extracellular vesicles using an in vivo whole organism approach

Boysen, Anders T. ; Whitehead, Bradley ; Stensballe, Allan ; Carnerup, Anna LU ; Nylander, Tommy LU and Nejsum, Peter (2020) In Biomedicines 8(7).
Abstract

In the last two decades, extracellular vesicles (EVs) from the three domains of life, Archaea, Bacteria and Eukaryotes, have gained increasing scientific attention. As such, the role of EVs in host-pathogen communication and immune modulation are being intensely investigated. Pivotal to EV research is the determination of how and where EVs are taken up by recipient cells and organs in vivo, which requires suitable tracking strategies including labelling. Labelling of EVs is often performed post-isolation which increases risks of non-specific labelling and the introduction of labelling artefacts. Here we exploited the inability of helminths to de novo synthesise fatty acids to enable labelling of EVs by whole organism uptake of... (More)

In the last two decades, extracellular vesicles (EVs) from the three domains of life, Archaea, Bacteria and Eukaryotes, have gained increasing scientific attention. As such, the role of EVs in host-pathogen communication and immune modulation are being intensely investigated. Pivotal to EV research is the determination of how and where EVs are taken up by recipient cells and organs in vivo, which requires suitable tracking strategies including labelling. Labelling of EVs is often performed post-isolation which increases risks of non-specific labelling and the introduction of labelling artefacts. Here we exploited the inability of helminths to de novo synthesise fatty acids to enable labelling of EVs by whole organism uptake of fluorescent lipid analogues and the subsequent incorporation in EVs. We showed uptake of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (DOPE-Rho) in Anisakis spp. and Trichuris suis larvae. EVs isolated from the supernatant of Anisakis spp. labelled with DOPE-Rho were characterised to assess the effects of labelling on size, structure and fluorescence of EVs. Fluorescent EVs were successfully taken up by the human macrophage cell line THP-1. This study, therefore, presents a novel staining method that can be utilized by the EV field in parasitology and potentially across multiple species.

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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cryo-EM, Extracellular vesicles, Helminth, Proteomics, Vesicle labelling, Vesicle tracking
in
Biomedicines
volume
8
issue
7
article number
213
publisher
MDPI AG
external identifiers
  • scopus:85089683373
ISSN
2227-9059
DOI
10.3390/BIOMEDICINES8070213
language
English
LU publication?
yes
id
07829161-02a6-4c76-a501-eb6489377c11
date added to LUP
2021-01-12 13:22:49
date last changed
2023-11-20 20:07:25
@article{07829161-02a6-4c76-a501-eb6489377c11,
  abstract     = {{<p>In the last two decades, extracellular vesicles (EVs) from the three domains of life, Archaea, Bacteria and Eukaryotes, have gained increasing scientific attention. As such, the role of EVs in host-pathogen communication and immune modulation are being intensely investigated. Pivotal to EV research is the determination of how and where EVs are taken up by recipient cells and organs in vivo, which requires suitable tracking strategies including labelling. Labelling of EVs is often performed post-isolation which increases risks of non-specific labelling and the introduction of labelling artefacts. Here we exploited the inability of helminths to de novo synthesise fatty acids to enable labelling of EVs by whole organism uptake of fluorescent lipid analogues and the subsequent incorporation in EVs. We showed uptake of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (DOPE-Rho) in Anisakis spp. and Trichuris suis larvae. EVs isolated from the supernatant of Anisakis spp. labelled with DOPE-Rho were characterised to assess the effects of labelling on size, structure and fluorescence of EVs. Fluorescent EVs were successfully taken up by the human macrophage cell line THP-1. This study, therefore, presents a novel staining method that can be utilized by the EV field in parasitology and potentially across multiple species.</p>}},
  author       = {{Boysen, Anders T. and Whitehead, Bradley and Stensballe, Allan and Carnerup, Anna and Nylander, Tommy and Nejsum, Peter}},
  issn         = {{2227-9059}},
  keywords     = {{Cryo-EM; Extracellular vesicles; Helminth; Proteomics; Vesicle labelling; Vesicle tracking}},
  language     = {{eng}},
  number       = {{7}},
  publisher    = {{MDPI AG}},
  series       = {{Biomedicines}},
  title        = {{Fluorescent labeling of helminth extracellular vesicles using an in vivo whole organism approach}},
  url          = {{http://dx.doi.org/10.3390/BIOMEDICINES8070213}},
  doi          = {{10.3390/BIOMEDICINES8070213}},
  volume       = {{8}},
  year         = {{2020}},
}