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The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils

Basophils are granulocytes involved in parasite immunity and allergic diseases, known for their potent secretion of type 2 cytokines. Identifying their functions has proven to be controversial due to their relative rarity and their complex lineage phenotype. Here, we show that the expression of baso...

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Autores principales: Pellefigues, Christophe, Mehta, Palak, Prout, Melanie Sarah, Naidoo, Karmella, Yumnam, Bibek, Chandler, Jodie, Chappell, Sally, Filbey, Kara, Camberis, Mali, Le Gros, Graham
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6746837/
https://www.ncbi.nlm.nih.gov/pubmed/31552058
http://dx.doi.org/10.3389/fimmu.2019.02143
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author Pellefigues, Christophe
Mehta, Palak
Prout, Melanie Sarah
Naidoo, Karmella
Yumnam, Bibek
Chandler, Jodie
Chappell, Sally
Filbey, Kara
Camberis, Mali
Le Gros, Graham
author_facet Pellefigues, Christophe
Mehta, Palak
Prout, Melanie Sarah
Naidoo, Karmella
Yumnam, Bibek
Chandler, Jodie
Chappell, Sally
Filbey, Kara
Camberis, Mali
Le Gros, Graham
author_sort Pellefigues, Christophe
collection PubMed
description Basophils are granulocytes involved in parasite immunity and allergic diseases, known for their potent secretion of type 2 cytokines. Identifying their functions has proven to be controversial due to their relative rarity and their complex lineage phenotype. Here, we show that the expression of basophils lineage markers CD200R3 and FcεRIα is highly variable in inflammatory settings and hinders basophils identification by flow cytometry across multiple disease states or tissues. Fluorophore-conjugated antibody staining of these lineage markers strongly activates basophil type 2 cytokine expression, and represents a potential bias for coculture or in vivo transfer experiments. The Basoph8 is a mouse model where basophils specifically express a strong fluorescent reporter and the Cre recombinase. Basophils can be identified and FACS sorted unambiguously by their expression of the enhanced yellow fluorescent protein (eYFP) in these mice. We show that the expression of the eYFP is robust in vivo during inflammation, and in vitro on living basophils for at least 72 h, including during the induction of anaphylactoid degranulation. We bred and characterized the Basoph8xiDTR mice, in which basophils specifically express eYFP and the simian diphtheria toxin receptor (DTR). This model enables basophils conditional depletion relatively specifically ex vivo and in vivo during allergic inflammation and their detection as eYFP+ cells. In conclusion, we report underappreciated benefits of the commercially available Basoph8 mice to study basophils function.
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spelling pubmed-67468372019-09-24 The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils Pellefigues, Christophe Mehta, Palak Prout, Melanie Sarah Naidoo, Karmella Yumnam, Bibek Chandler, Jodie Chappell, Sally Filbey, Kara Camberis, Mali Le Gros, Graham Front Immunol Immunology Basophils are granulocytes involved in parasite immunity and allergic diseases, known for their potent secretion of type 2 cytokines. Identifying their functions has proven to be controversial due to their relative rarity and their complex lineage phenotype. Here, we show that the expression of basophils lineage markers CD200R3 and FcεRIα is highly variable in inflammatory settings and hinders basophils identification by flow cytometry across multiple disease states or tissues. Fluorophore-conjugated antibody staining of these lineage markers strongly activates basophil type 2 cytokine expression, and represents a potential bias for coculture or in vivo transfer experiments. The Basoph8 is a mouse model where basophils specifically express a strong fluorescent reporter and the Cre recombinase. Basophils can be identified and FACS sorted unambiguously by their expression of the enhanced yellow fluorescent protein (eYFP) in these mice. We show that the expression of the eYFP is robust in vivo during inflammation, and in vitro on living basophils for at least 72 h, including during the induction of anaphylactoid degranulation. We bred and characterized the Basoph8xiDTR mice, in which basophils specifically express eYFP and the simian diphtheria toxin receptor (DTR). This model enables basophils conditional depletion relatively specifically ex vivo and in vivo during allergic inflammation and their detection as eYFP+ cells. In conclusion, we report underappreciated benefits of the commercially available Basoph8 mice to study basophils function. Frontiers Media S.A. 2019-09-10 /pmc/articles/PMC6746837/ /pubmed/31552058 http://dx.doi.org/10.3389/fimmu.2019.02143 Text en Copyright © 2019 Pellefigues, Mehta, Prout, Naidoo, Yumnam, Chandler, Chappell, Filbey, Camberis and Le Gros. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Pellefigues, Christophe
Mehta, Palak
Prout, Melanie Sarah
Naidoo, Karmella
Yumnam, Bibek
Chandler, Jodie
Chappell, Sally
Filbey, Kara
Camberis, Mali
Le Gros, Graham
The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils
title The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils
title_full The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils
title_fullStr The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils
title_full_unstemmed The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils
title_short The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils
title_sort basoph8 mice enable an unbiased detection and a conditional depletion of basophils
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6746837/
https://www.ncbi.nlm.nih.gov/pubmed/31552058
http://dx.doi.org/10.3389/fimmu.2019.02143
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