Cargando…

Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells

Analysis of chemokine receptor, and atypical chemokine receptor, expression is frequently hampered by the lack of availability of high‐quality antibodies and the species specificity of those that are available. We have previously described methodology utilizing Alexa‐Fluor‐labeled chemokine ligands...

Descripción completa

Detalles Bibliográficos
Autores principales: Hansell, Christopher A.H., Love, Samantha, Pingen, Marieke, Wilson, Gillian J., MacLeod, Megan, Graham, Gerard J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8638673/
https://www.ncbi.nlm.nih.gov/pubmed/32114694
http://dx.doi.org/10.1002/eji.201948374
_version_ 1784608989126852608
author Hansell, Christopher A.H.
Love, Samantha
Pingen, Marieke
Wilson, Gillian J.
MacLeod, Megan
Graham, Gerard J.
author_facet Hansell, Christopher A.H.
Love, Samantha
Pingen, Marieke
Wilson, Gillian J.
MacLeod, Megan
Graham, Gerard J.
author_sort Hansell, Christopher A.H.
collection PubMed
description Analysis of chemokine receptor, and atypical chemokine receptor, expression is frequently hampered by the lack of availability of high‐quality antibodies and the species specificity of those that are available. We have previously described methodology utilizing Alexa‐Fluor‐labeled chemokine ligands as versatile reagents to detect receptor expression. Previously this has been limited to hematopoietic cells and methodology for assessing expression of receptors on stromal cells has been lacking. Among chemokine receptors, the ones most frequently expressed on stromal cells belong to the atypical chemokine receptor subfamily. These receptors do not signal in the classic sense in response to ligand but scavenge their ligands and degrade them and thus sculpt in vivo chemokine gradients. Here, we demonstrate the ability to use either intratracheal or intravenous, Alexa‐Fluor‐labeled chemokine administration to detect stromal cell populations expressing the atypical chemokine receptor ACKR2. Using this methodology, we demonstrate, for the first time, expression of ACKR2 on blood endothelial cells. This observation sets the lung aside from other tissues in which ACKR2 is exclusively expressed on lymphatic endothelial cells and suggest unique roles for ACKR2 in the pulmonary environment.
format Online
Article
Text
id pubmed-8638673
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-86386732021-12-09 Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells Hansell, Christopher A.H. Love, Samantha Pingen, Marieke Wilson, Gillian J. MacLeod, Megan Graham, Gerard J. Eur J Immunol Tissue immunology and leukocyte trafficking Analysis of chemokine receptor, and atypical chemokine receptor, expression is frequently hampered by the lack of availability of high‐quality antibodies and the species specificity of those that are available. We have previously described methodology utilizing Alexa‐Fluor‐labeled chemokine ligands as versatile reagents to detect receptor expression. Previously this has been limited to hematopoietic cells and methodology for assessing expression of receptors on stromal cells has been lacking. Among chemokine receptors, the ones most frequently expressed on stromal cells belong to the atypical chemokine receptor subfamily. These receptors do not signal in the classic sense in response to ligand but scavenge their ligands and degrade them and thus sculpt in vivo chemokine gradients. Here, we demonstrate the ability to use either intratracheal or intravenous, Alexa‐Fluor‐labeled chemokine administration to detect stromal cell populations expressing the atypical chemokine receptor ACKR2. Using this methodology, we demonstrate, for the first time, expression of ACKR2 on blood endothelial cells. This observation sets the lung aside from other tissues in which ACKR2 is exclusively expressed on lymphatic endothelial cells and suggest unique roles for ACKR2 in the pulmonary environment. John Wiley and Sons Inc. 2020-03-17 2020-05 /pmc/articles/PMC8638673/ /pubmed/32114694 http://dx.doi.org/10.1002/eji.201948374 Text en © 2020 The Authors. European Journal of Immunology published by WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Tissue immunology and leukocyte trafficking
Hansell, Christopher A.H.
Love, Samantha
Pingen, Marieke
Wilson, Gillian J.
MacLeod, Megan
Graham, Gerard J.
Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells
title Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells
title_full Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells
title_fullStr Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells
title_full_unstemmed Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells
title_short Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells
title_sort analysis of lung stromal expression of the atypical chemokine receptor ackr2 reveals unanticipated expression in murine blood endothelial cells
topic Tissue immunology and leukocyte trafficking
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8638673/
https://www.ncbi.nlm.nih.gov/pubmed/32114694
http://dx.doi.org/10.1002/eji.201948374
work_keys_str_mv AT hansellchristopherah analysisoflungstromalexpressionoftheatypicalchemokinereceptorackr2revealsunanticipatedexpressioninmurinebloodendothelialcells
AT lovesamantha analysisoflungstromalexpressionoftheatypicalchemokinereceptorackr2revealsunanticipatedexpressioninmurinebloodendothelialcells
AT pingenmarieke analysisoflungstromalexpressionoftheatypicalchemokinereceptorackr2revealsunanticipatedexpressioninmurinebloodendothelialcells
AT wilsongillianj analysisoflungstromalexpressionoftheatypicalchemokinereceptorackr2revealsunanticipatedexpressioninmurinebloodendothelialcells
AT macleodmegan analysisoflungstromalexpressionoftheatypicalchemokinereceptorackr2revealsunanticipatedexpressioninmurinebloodendothelialcells
AT grahamgerardj analysisoflungstromalexpressionoftheatypicalchemokinereceptorackr2revealsunanticipatedexpressioninmurinebloodendothelialcells