Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 Functions
Chemokines are essential guidance cues orchestrating cell migration in health and disease. Cognate chemokine receptors sense chemokine gradients over short distances to coordinate directional cell locomotion. The chemokines CCL19 and CCL21 are essential for recruiting CCR7-expressing dendritic cells...
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doaj-3b3090c3db654ed8b43ba141ec284a002020-11-24T21:28:54ZengMDPI AGInternational Journal of Molecular Sciences1422-00672018-12-011912387610.3390/ijms19123876ijms19123876Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 FunctionsVladimir Purvanov0Christoph Matti1Guerric P. B. Samson2Ilona Kindinger3Daniel F. Legler4Biotechnology Institute Thurgau (BITg) at the University of Konstanz, CH-8280 Kreuzlingen, SwitzerlandBiotechnology Institute Thurgau (BITg) at the University of Konstanz, CH-8280 Kreuzlingen, SwitzerlandBiotechnology Institute Thurgau (BITg) at the University of Konstanz, CH-8280 Kreuzlingen, SwitzerlandBiotechnology Institute Thurgau (BITg) at the University of Konstanz, CH-8280 Kreuzlingen, SwitzerlandBiotechnology Institute Thurgau (BITg) at the University of Konstanz, CH-8280 Kreuzlingen, SwitzerlandChemokines are essential guidance cues orchestrating cell migration in health and disease. Cognate chemokine receptors sense chemokine gradients over short distances to coordinate directional cell locomotion. The chemokines CCL19 and CCL21 are essential for recruiting CCR7-expressing dendritic cells bearing pathogen-derived antigens and lymphocytes to lymph nodes, where the two cell types meet to launch an adaptive immune response against the invading pathogen. CCR7-expressing cancer cells are also recruited by CCL19 and CCL21 to metastasize in lymphoid organs. In contrast, atypical chemokine receptors (ACKRs) do not transmit signals required for cell locomotion but scavenge chemokines. ACKR4 is crucial for internalizing and degrading CCL19 and CCL21 to establish local gradients, which are sensed by CCR7-expressing cells. Here, we describe the production of fluorescently tagged chemokines by fusing CCL19 and CCL21 to monomeric red fluorescent protein (mRFP). We show that purified CCL19-mRFP and CCL21-mRFP are versatile and powerful tools to study CCR7 and ACKR4 functions, such as receptor trafficking and chemokine scavenging, in a spatiotemporal fashion. We demonstrate that fluorescently tagged CCL19 and CCL21 permit the visualization and quantification of chemokine gradients in real time, while CCR7-expressing leukocytes and cancer cells sense the guidance cues and migrate along the chemokine gradients.https://www.mdpi.com/1422-0067/19/12/3876fluorescent chemokineschemokine receptorsCCL19CCL21CCR7ACKR4cell migrationleukocytescancer metastasis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Vladimir Purvanov Christoph Matti Guerric P. B. Samson Ilona Kindinger Daniel F. Legler |
spellingShingle |
Vladimir Purvanov Christoph Matti Guerric P. B. Samson Ilona Kindinger Daniel F. Legler Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 Functions International Journal of Molecular Sciences fluorescent chemokines chemokine receptors CCL19 CCL21 CCR7 ACKR4 cell migration leukocytes cancer metastasis |
author_facet |
Vladimir Purvanov Christoph Matti Guerric P. B. Samson Ilona Kindinger Daniel F. Legler |
author_sort |
Vladimir Purvanov |
title |
Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 Functions |
title_short |
Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 Functions |
title_full |
Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 Functions |
title_fullStr |
Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 Functions |
title_full_unstemmed |
Fluorescently Tagged CCL19 and CCL21 to Monitor CCR7 and ACKR4 Functions |
title_sort |
fluorescently tagged ccl19 and ccl21 to monitor ccr7 and ackr4 functions |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2018-12-01 |
description |
Chemokines are essential guidance cues orchestrating cell migration in health and disease. Cognate chemokine receptors sense chemokine gradients over short distances to coordinate directional cell locomotion. The chemokines CCL19 and CCL21 are essential for recruiting CCR7-expressing dendritic cells bearing pathogen-derived antigens and lymphocytes to lymph nodes, where the two cell types meet to launch an adaptive immune response against the invading pathogen. CCR7-expressing cancer cells are also recruited by CCL19 and CCL21 to metastasize in lymphoid organs. In contrast, atypical chemokine receptors (ACKRs) do not transmit signals required for cell locomotion but scavenge chemokines. ACKR4 is crucial for internalizing and degrading CCL19 and CCL21 to establish local gradients, which are sensed by CCR7-expressing cells. Here, we describe the production of fluorescently tagged chemokines by fusing CCL19 and CCL21 to monomeric red fluorescent protein (mRFP). We show that purified CCL19-mRFP and CCL21-mRFP are versatile and powerful tools to study CCR7 and ACKR4 functions, such as receptor trafficking and chemokine scavenging, in a spatiotemporal fashion. We demonstrate that fluorescently tagged CCL19 and CCL21 permit the visualization and quantification of chemokine gradients in real time, while CCR7-expressing leukocytes and cancer cells sense the guidance cues and migrate along the chemokine gradients. |
topic |
fluorescent chemokines chemokine receptors CCL19 CCL21 CCR7 ACKR4 cell migration leukocytes cancer metastasis |
url |
https://www.mdpi.com/1422-0067/19/12/3876 |
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