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

Biomedicines
Anders Toftegaard BoysenPeter Nejsum

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, presen...Continue Reading

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Citations

Jul 11, 2021·Molecular Immunology·Claire Drurey, Rick M Maizels

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Methods Mentioned

BETA
PMA
fluorescence microscopy

Software Mentioned

BLASTP
ImageJ
Nanosight NTA
Blast2Go
Maxquant

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