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dc.contributor.authorVan, Andre Pen_UK
dc.contributor.authorAlvarez de Haro, Neilaen_UK
dc.contributor.authorBron, James Een_UK
dc.contributor.authorDesbois, Andrew Pen_UK
dc.description.abstractNeutrophils release nuclear chromatin decorated with antimicrobial proteins into the extracellular milieu as an innate immune defence mechanism to counter invading microbes. These chromatin structures, called extracellular traps (ETs) and released by a process called NETosis, have been detected in mammals, certain invertebrates and some fish species, including fathead minnow, zebrafish, common carp, turbot, sole and barramundi. However, there have been no previous studies of ETs in the Salmonidae. ETs are released in response to chemical and biological stimuli, but observations from different fish species are inconsistent, particularly regarding the potency of various inducers and inhibitors. Thus, this present study aimed to describe ET release in a salmonid (rainbow trout, Oncorhynchus mykiss (Walbaum, 1792)) and uncover the inducers and inhibitors that can control this response. Highly enriched suspensions of polymorphonuclear cells (PMNs; mainly neutrophils) were prepared from head kidney tissues by a triple-layer Percoll gradient technique. ET structures were visualised in PMN-enriched suspensions through staining of the chromatin with nucleic acid-specific dyes and immunocytochemical probing of characteristic proteins expected to decorate the structure. ET release was quantified after incubation with inducers and inhibitors known to affect this response in other organisms. Structures resembling ETs stained positively with SYTOX Green (a stain specific for nucleic acid) while immunocytochemistry was used to detect neutrophil elastase, myeloperoxidase and H2A histone in the structures, which are diagnostic proteinaceous markers of ETs. Consistent with other studies on mammals and some fish species, calcium ionophore and flagellin were potent inducers of ETs, while cytochalasin D inhibited NETosis. Phorbol 12-myristate 13-acetate (PMA), used commonly to induce ETs, exerted only weak stimulatory activity, while heat-killed bacteria and lipopolysaccharide did not induce ET release. Unexpectedly, the ET-inhibitor diphenyleneiodonium chloride acted as an inducer of ET release, an observation not reported elsewhere. Taken together, these data confirm for the first time that ETs are released by salmonid PMNs and compounds useful for manipulating NETosis were identified, thus providing a platform for further studies to explore the role of this mechanism in fish immunity. This new knowledge provides a foundation for translation to farm settings, since manipulation of the innate immune response offers a potential alternative to the use of antibiotics to mitigate against microbial infections, particularly for pathogens where protection by vaccination has yet to be realised.en_UK
dc.publisherElsevier BVen_UK
dc.relationVan AP, Alvarez de Haro N, Bron JE & Desbois AP (2020) Chromatin extracellular trap release in rainbow trout, Oncorhynchus mykiss (Walbaum, 1792). Fish and Shellfish Immunology, 99, pp. 227-238.
dc.rightsThis is an open access article distributed under the terms of the Creative Commons CC-BY license (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. You are not required to obtain permission to reuse this article.en_UK
dc.subjectInnate immunityen_UK
dc.subjectNeutrophil extracellular traps (NETs)en_UK
dc.titleChromatin extracellular trap release in rainbow trout, Oncorhynchus mykiss (Walbaum, 1792)en_UK
dc.typeJournal Articleen_UK
dc.citation.jtitleFish and Shellfish Immunologyen_UK
dc.type.statusVoR - Version of Recorden_UK
dc.contributor.funderBBSRC Biotechnology and Biological Sciences Research Councilen_UK
dc.contributor.affiliationInstitute of Aquacultureen_UK
dc.contributor.affiliationInstitute of Aquacultureen_UK
dc.contributor.affiliationInstitute of Aquacultureen_UK
dc.relation.funderprojectThe role of chromatin extracellular traps in host defence of fish against pathogensen_UK
Appears in Collections:Aquaculture Journal Articles

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