Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach

Boysen, Anders T.; Whitehead, Bradley; Stensballe, Allan; Carnerup, Anna; Nylander, Tommy; Nejsum, Peter

doi:10.3390/biomedicines8070213

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Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach

^1,†

^1,†

³,

and

^1,4,*

Department of Clinical Medicine, Aarhus University, Aarhus 8200, Denmark

Department of Health Science and Technology, Aalborg University, Aalborg 9100, Denmark

Department of Chemistry, Physical Chemistry, Lund University, Lund 210 00, Sweden

⁴

Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Melbourne 3010, Australia

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Biomedicines 2020, 8(7), 213; https://doi.org/10.3390/biomedicines8070213

Received: 18 June 2020 / Revised: 9 July 2020 / Accepted: 10 July 2020 / Published: 14 July 2020

(This article belongs to the Special Issue The Role of Extracellular Vesicles in Pathogenesis and Therapeutic Strategies)

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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 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. View Full-Text

Keywords: extracellular vesicles; vesicle labelling; vesicle tracking; helminth; proteomics; Cryo–EM

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MDPI and ACS Style

Boysen, A.T.; Whitehead, B.; Stensballe, A.; Carnerup, A.; Nylander, T.; Nejsum, P. Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach. Biomedicines 2020, 8, 213. https://doi.org/10.3390/biomedicines8070213

AMA Style

Boysen AT, Whitehead B, Stensballe A, Carnerup A, Nylander T, Nejsum P. Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach. Biomedicines. 2020; 8(7):213. https://doi.org/10.3390/biomedicines8070213

Chicago/Turabian Style

Boysen, Anders T., Bradley Whitehead, Allan Stensballe, Anna Carnerup, Tommy Nylander, and Peter Nejsum. 2020. "Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach" Biomedicines 8, no. 7: 213. https://doi.org/10.3390/biomedicines8070213

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Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach

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