Analysis of lung stromal expression of the atypical chemokine receptor ACKR2 reveals unanticipated expression in murine blood endothelial cells
| Autor: | Gerard J. Graham, Megan K. L. MacLeod, Gillian J. Wilson, Samantha Love, Christopher A.H. Hansell, Marieke Pingen |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
ACKR2 Pulmonary immunity Chemokine Stromal cell government.form_of_government Receptor expression Immunology Gene Expression Blood endothelial cells Mice 03 medical and health sciences Chemokine receptor 0302 clinical medicine Animals Immunology and Allergy Basic Receptor Tissue immunology and leukocyte trafficking Lung Fluorescent Dyes Mice Knockout Staining and Labeling biology Endothelial Cells Carbocyanines Epithelial Cell Adhesion Molecule Flow Cytometry 3. Good health Cell biology Lymphatic Endothelium Haematopoiesis 030104 developmental biology Alexa‐Fluor labeling biology.protein government Research Article|Basic Receptors Chemokine Stromal Cells Antibody Research Article 030215 immunology |
| Zdroj: | European Journal of Immunology |
| ISSN: | 1521-4141 0014-2980 |
| Popis: | 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. Fluorescent‐CCL22 administration in vivo reveals pulmonary ACKR2 expression patterns. Intratracheal administration labels fibroblasts and some endothelial cells, while intravenous administration labels vascular endothelial cells. Our data provide the first evidence of ACKR2 expression on vascular endothelial cells. This is a versatile method applicable to analyzing other chemokine receptors in vivo. |
| Databáze: | OpenAIRE |
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