MIF interacts with CXCR7 to promote receptor internalization, ERK1/2 and ZAP-70 signaling, and lymphocyte chemotaxis

Setareh Alampour-Rajabi; Omar El Bounkari; Antal Rot; Gerhard Müller-Newen; Françoise Bachelerie; Meinrad Gawaz; Christian Weber; Andreas Schober; Jürgen Bernhagen

doi:10.1096/fj.15-273904

MIF interacts with CXCR7 to promote receptor internalization, ERK1/2 and ZAP-70 signaling, and lymphocyte chemotaxis

Setareh Alampour-Rajabi, Omar El Bounkari, Antal Rot, Gerhard Müller-Newen, Françoise Bachelerie, Meinrad Gawaz, Christian Weber, Andreas Schober, Jürgen Bernhagen

Research output: Contribution to journal › Article › peer-review

Abstract

Macrophage migration-inhibitory factor (MIF) is a pleiotropic cytokine with chemokine-like functions and is a mediator in numerous inflammatory conditions. Depending on the context, MIF signals through 1 or more of its receptors cluster of differentiation (CD)74, CXC-motif chemokine receptor (CXCR)2, and CXCR4. In addition, heteromeric receptor complexes have been identified. We characterized the atypical chemokine receptor CXCR7 as a novel receptor for MIF. MIF promoted human CXCR7 internalization up to 40%, peaking at 50-400 nM and 30 min, but CXCR7 internalization by MIF was not dependent on CXCR4. Yet, by coimmunoprecipitation, fluorescence microscopy, and a proximity ligation assay, CXCR7 was found to engage in MIF receptor complexes with CXCR4 and CD74, both after ectopic overexpression and in endogenous conditions in a human B-cell line. Receptor competition binding and coimmunoprecipitation studies combined with sulfo-SBED-biotin-transfer provided evidence for a direct interaction between MIF and CXCR7. Finally, we demonstrated MIF/CXCR7-mediated functional responses. Blockade of CXCR7 suppressed MIF-mediated ERK- and zeta-chain-associated protein kinase (ZAP)-70 activation (from 2.1- to 1.2-fold and from 2.5- to 1.6-fold, respectively) and fully abrogated primary murine B-cell chemotaxis triggered by MIF, but not by CXCL12. B cells from Cxcr7(-/-) mice exhibited an ablated transmigration response to MIF, indicating that CXCR7 is essential for MIF-promoted B-cell migration. Our findings provide biochemical and functional evidence that MIF is an alternative ligand of CXCR7 and suggest a functional role of the MIF-CXCR7 axis in B-lymphocyte migration.

Original language	English
Pages (from-to)	4497-4511
Number of pages	15
Journal	The FASEB Journal
Volume	29
Issue number	11
DOIs	https://doi.org/10.1096/fj.15-273904
Publication status	Published - Nov 2015

Keywords

Animals
B-Lymphocytes
Cell Line, Tumor
Chemokine CXCL12
Chemotaxis
Humans
Intramolecular Oxidoreductases
MAP Kinase Signaling System
Macrophage Migration-Inhibitory Factors
Mice
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
Receptors, CXCR
Receptors, CXCR4
ZAP-70 Protein-Tyrosine Kinase

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10.1096/fj.15-273904

Cite this

@article{a3c8aa2911254afca39d79652853ad2e,

title = "MIF interacts with CXCR7 to promote receptor internalization, ERK1/2 and ZAP-70 signaling, and lymphocyte chemotaxis",

abstract = "Macrophage migration-inhibitory factor (MIF) is a pleiotropic cytokine with chemokine-like functions and is a mediator in numerous inflammatory conditions. Depending on the context, MIF signals through 1 or more of its receptors cluster of differentiation (CD)74, CXC-motif chemokine receptor (CXCR)2, and CXCR4. In addition, heteromeric receptor complexes have been identified. We characterized the atypical chemokine receptor CXCR7 as a novel receptor for MIF. MIF promoted human CXCR7 internalization up to 40%, peaking at 50-400 nM and 30 min, but CXCR7 internalization by MIF was not dependent on CXCR4. Yet, by coimmunoprecipitation, fluorescence microscopy, and a proximity ligation assay, CXCR7 was found to engage in MIF receptor complexes with CXCR4 and CD74, both after ectopic overexpression and in endogenous conditions in a human B-cell line. Receptor competition binding and coimmunoprecipitation studies combined with sulfo-SBED-biotin-transfer provided evidence for a direct interaction between MIF and CXCR7. Finally, we demonstrated MIF/CXCR7-mediated functional responses. Blockade of CXCR7 suppressed MIF-mediated ERK- and zeta-chain-associated protein kinase (ZAP)-70 activation (from 2.1- to 1.2-fold and from 2.5- to 1.6-fold, respectively) and fully abrogated primary murine B-cell chemotaxis triggered by MIF, but not by CXCL12. B cells from Cxcr7(-/-) mice exhibited an ablated transmigration response to MIF, indicating that CXCR7 is essential for MIF-promoted B-cell migration. Our findings provide biochemical and functional evidence that MIF is an alternative ligand of CXCR7 and suggest a functional role of the MIF-CXCR7 axis in B-lymphocyte migration.",

keywords = "Animals, B-Lymphocytes, Cell Line, Tumor, Chemokine CXCL12, Chemotaxis, Humans, Intramolecular Oxidoreductases, MAP Kinase Signaling System, Macrophage Migration-Inhibitory Factors, Mice, Mitogen-Activated Protein Kinase 1, Mitogen-Activated Protein Kinase 3, Receptors, CXCR, Receptors, CXCR4, ZAP-70 Protein-Tyrosine Kinase",

author = "Setareh Alampour-Rajabi and {El Bounkari}, Omar and Antal Rot and Gerhard M{\"u}ller-Newen and Fran{\c c}oise Bachelerie and Meinrad Gawaz and Christian Weber and Andreas Schober and J{\"u}rgen Bernhagen",

note = "{\textcopyright} FASEB.",

year = "2015",

month = nov,

doi = "10.1096/fj.15-273904",

language = "English",

volume = "29",

pages = "4497--4511",

journal = "The FASEB Journal",

issn = "0892-6638",

publisher = "FASEB",

number = "11",

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TY - JOUR

T1 - MIF interacts with CXCR7 to promote receptor internalization, ERK1/2 and ZAP-70 signaling, and lymphocyte chemotaxis

AU - Alampour-Rajabi, Setareh

AU - El Bounkari, Omar

AU - Rot, Antal

AU - Müller-Newen, Gerhard

AU - Bachelerie, Françoise

AU - Gawaz, Meinrad

AU - Weber, Christian

AU - Schober, Andreas

AU - Bernhagen, Jürgen

N1 - © FASEB.

PY - 2015/11

Y1 - 2015/11

N2 - Macrophage migration-inhibitory factor (MIF) is a pleiotropic cytokine with chemokine-like functions and is a mediator in numerous inflammatory conditions. Depending on the context, MIF signals through 1 or more of its receptors cluster of differentiation (CD)74, CXC-motif chemokine receptor (CXCR)2, and CXCR4. In addition, heteromeric receptor complexes have been identified. We characterized the atypical chemokine receptor CXCR7 as a novel receptor for MIF. MIF promoted human CXCR7 internalization up to 40%, peaking at 50-400 nM and 30 min, but CXCR7 internalization by MIF was not dependent on CXCR4. Yet, by coimmunoprecipitation, fluorescence microscopy, and a proximity ligation assay, CXCR7 was found to engage in MIF receptor complexes with CXCR4 and CD74, both after ectopic overexpression and in endogenous conditions in a human B-cell line. Receptor competition binding and coimmunoprecipitation studies combined with sulfo-SBED-biotin-transfer provided evidence for a direct interaction between MIF and CXCR7. Finally, we demonstrated MIF/CXCR7-mediated functional responses. Blockade of CXCR7 suppressed MIF-mediated ERK- and zeta-chain-associated protein kinase (ZAP)-70 activation (from 2.1- to 1.2-fold and from 2.5- to 1.6-fold, respectively) and fully abrogated primary murine B-cell chemotaxis triggered by MIF, but not by CXCL12. B cells from Cxcr7(-/-) mice exhibited an ablated transmigration response to MIF, indicating that CXCR7 is essential for MIF-promoted B-cell migration. Our findings provide biochemical and functional evidence that MIF is an alternative ligand of CXCR7 and suggest a functional role of the MIF-CXCR7 axis in B-lymphocyte migration.

AB - Macrophage migration-inhibitory factor (MIF) is a pleiotropic cytokine with chemokine-like functions and is a mediator in numerous inflammatory conditions. Depending on the context, MIF signals through 1 or more of its receptors cluster of differentiation (CD)74, CXC-motif chemokine receptor (CXCR)2, and CXCR4. In addition, heteromeric receptor complexes have been identified. We characterized the atypical chemokine receptor CXCR7 as a novel receptor for MIF. MIF promoted human CXCR7 internalization up to 40%, peaking at 50-400 nM and 30 min, but CXCR7 internalization by MIF was not dependent on CXCR4. Yet, by coimmunoprecipitation, fluorescence microscopy, and a proximity ligation assay, CXCR7 was found to engage in MIF receptor complexes with CXCR4 and CD74, both after ectopic overexpression and in endogenous conditions in a human B-cell line. Receptor competition binding and coimmunoprecipitation studies combined with sulfo-SBED-biotin-transfer provided evidence for a direct interaction between MIF and CXCR7. Finally, we demonstrated MIF/CXCR7-mediated functional responses. Blockade of CXCR7 suppressed MIF-mediated ERK- and zeta-chain-associated protein kinase (ZAP)-70 activation (from 2.1- to 1.2-fold and from 2.5- to 1.6-fold, respectively) and fully abrogated primary murine B-cell chemotaxis triggered by MIF, but not by CXCL12. B cells from Cxcr7(-/-) mice exhibited an ablated transmigration response to MIF, indicating that CXCR7 is essential for MIF-promoted B-cell migration. Our findings provide biochemical and functional evidence that MIF is an alternative ligand of CXCR7 and suggest a functional role of the MIF-CXCR7 axis in B-lymphocyte migration.

KW - Animals

KW - B-Lymphocytes

KW - Cell Line, Tumor

KW - Chemokine CXCL12

KW - Chemotaxis

KW - Humans

KW - Intramolecular Oxidoreductases

KW - MAP Kinase Signaling System

KW - Macrophage Migration-Inhibitory Factors

KW - Mice

KW - Mitogen-Activated Protein Kinase 1

KW - Mitogen-Activated Protein Kinase 3

KW - Receptors, CXCR

KW - Receptors, CXCR4

KW - ZAP-70 Protein-Tyrosine Kinase

U2 - 10.1096/fj.15-273904

DO - 10.1096/fj.15-273904

M3 - Article

C2 - 26139098

SN - 0892-6638

VL - 29

SP - 4497

EP - 4511

JO - The FASEB Journal

JF - The FASEB Journal

IS - 11

ER -