Exosomal release of the virus-encoded chemokine receptor US28 contributes to chemokine scavenging

Maarten P. Bebelman; Irfan M. Setiawan; Nick D. Bergkamp; Jeffrey R. van Senten; Caitrin Crudden; Jan Paul M. Bebelman; Frederik J. Verweij; Guillaume van Niel; Marco Siderius; D. Michiel Pegtel; Martine J. Smit

doi:10.1016/j.isci.2023.107412

Exosomal release of the virus-encoded chemokine receptor US28 contributes to chemokine scavenging

Maarten P. Bebelman, Irfan M. Setiawan, Nick D. Bergkamp, Jeffrey R. van Senten, Caitrin Crudden, Jan Paul M. Bebelman, Frederik J. Verweij, Guillaume van Niel, Marco Siderius, D. Michiel Pegtel, Martine J. Smit^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

The human cytomegalovirus (HCMV)-encoded chemokine receptor US28 contributes to various aspects of the viral life cycle and promotes immune evasion by scavenging chemokines from the microenvironment of HCMV-infected cells. In contrast to the plasma membrane localization of most human chemokine receptors, US28 has a predominant intracellular localization. In this study, we used immunofluorescence and electron microscopy to determine the localization of US28 upon exogenous expression, as well as in HCMV-infected cells. We observed that US28 localizes to late endosomal compartments called multivesicular bodies (MVBs), where it is sorted in intraluminal vesicles. Live-cell total internal reflection fluorescence (TIRF) microscopy revealed that US28-containing MVBs can fuse with the plasma membrane, resulting in the secretion of US28 on exosomes. Exosomal US28 binds the chemokines CX₃CL1 and CCL5, and US28-containing exosomes inhibited the CX₃CL1-CX₃CR1 signaling axis. These findings suggest that exosomal release of US28 contributes to chemokine scavenging and immune evasion by HCMV.

Original language	English
Article number	107412
Pages (from-to)	1-16
Number of pages	17
Journal	iScience
Volume	26
Issue number	8
Early online date	18 Jul 2023
DOIs	https://doi.org/10.1016/j.isci.2023.107412
Publication status	Published - 18 Aug 2023

Bibliographical note

Funding Information:
We thank R.J. Stanton (Cardiff University, Cardiff, UK) for sharing the HCMV Merlin BAC and HFFF-Tet cells. We greatly acknowledge the AO|2M microscopy core platform of VU University Medical Center Amsterdam and the Center for Neurogenetics and Cognitive Research imaging platform of the Neuroscience Campus Amsterdam for imaging support. We greatly acknowledge the Cell and Tissue Imaging Center (PICT-IBiSA) #Institut Curie for electron microscopy. This work was funded by a grant from the Dutch Research Council (NWO: Vici grant 016.140.657) to M.J. Smit; an NWO–Amsterdam Institute for Molecules, Medicines, and Systems STAR Graduate Program grant (022.005.031) and a Cancer Center Amsterdam travel grant to M.P. Bebelman; A European Molecular Biology Organization grant (EMBO ALTF 1383-2014) and a Fondation ARC pour la Recherché sur le Cancer fellowship (PJA 20161204808) to F.J. Verweij; and a Fondation pour la Recherche Médicale grant (AJE20160635884) and an Institut National Du Cancer grant (INCA N°2019-125 PLBIO19-059) to F.J. Verweij. and G. Van Niel. Conceptualization, M.P.B. D.M.P. M.S. and M.J.S.; Formal Analysis: M.P.B. I.M.S. and N.D.B.; Investigation: M.P.B. I.M.S. N.D.B. J.R.v.S. and J.P.B.; Writing – Original Draft: M.P.B.; Writing – Review & Editing: M.P.B. N.D.B. J.R.v.S. C.C. J.P.B. D.M.P. M.S. and M.J.S.; Visualization: M.P.B.; Supervision: C.C. F.J.V. M.S. D.M.P. and M.J.S.; Funding Acquisition: M.P.B. F.J.V. G.v.N. and M.J.S. M.J.S. is inventor of patent WO2019151865A1 describing VUN100.

Funding Information:
This work was funded by a grant from the Dutch Research Council (NWO: Vici grant 016.140.657 ) to M.J. Smit; an NWO–Amsterdam Institute for Molecules, Medicines, and Systems STAR Graduate Program grant ( 022.005.031 ) and a Cancer Center Amsterdam travel grant to M.P. Bebelman; A European Molecular Biology Organization grant ( EMBO ALTF 1383-2014 ) and a Fondation ARC pour la Recherché sur le Cancer fellowship (PJA 20161204808 ) to F.J. Verweij; and a Fondation pour la Recherche Médicale grant ( AJE20160635884 ) and an Institut National Du Cancer grant (INCA N°2019-125 PLBIO19-059 ) to F.J. Verweij. and G. Van Niel.

Publisher Copyright:
© 2023 The Author(s)

Keywords

Cell biology
Immunology

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10.1016/j.isci.2023.107412

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@article{fafa89758ade4d10becc73d32333a94c,

title = "Exosomal release of the virus-encoded chemokine receptor US28 contributes to chemokine scavenging",

abstract = "The human cytomegalovirus (HCMV)-encoded chemokine receptor US28 contributes to various aspects of the viral life cycle and promotes immune evasion by scavenging chemokines from the microenvironment of HCMV-infected cells. In contrast to the plasma membrane localization of most human chemokine receptors, US28 has a predominant intracellular localization. In this study, we used immunofluorescence and electron microscopy to determine the localization of US28 upon exogenous expression, as well as in HCMV-infected cells. We observed that US28 localizes to late endosomal compartments called multivesicular bodies (MVBs), where it is sorted in intraluminal vesicles. Live-cell total internal reflection fluorescence (TIRF) microscopy revealed that US28-containing MVBs can fuse with the plasma membrane, resulting in the secretion of US28 on exosomes. Exosomal US28 binds the chemokines CX3CL1 and CCL5, and US28-containing exosomes inhibited the CX3CL1-CX3CR1 signaling axis. These findings suggest that exosomal release of US28 contributes to chemokine scavenging and immune evasion by HCMV.",

keywords = "Cell biology, Immunology",

author = "Bebelman, {Maarten P.} and Setiawan, {Irfan M.} and Bergkamp, {Nick D.} and {van Senten}, {Jeffrey R.} and Caitrin Crudden and Bebelman, {Jan Paul M.} and Verweij, {Frederik J.} and {van Niel}, Guillaume and Marco Siderius and Pegtel, {D. Michiel} and Smit, {Martine J.}",

note = "Funding Information: We thank R.J. Stanton (Cardiff University, Cardiff, UK) for sharing the HCMV Merlin BAC and HFFF-Tet cells. We greatly acknowledge the AO|2M microscopy core platform of VU University Medical Center Amsterdam and the Center for Neurogenetics and Cognitive Research imaging platform of the Neuroscience Campus Amsterdam for imaging support. We greatly acknowledge the Cell and Tissue Imaging Center (PICT-IBiSA) #Institut Curie for electron microscopy. This work was funded by a grant from the Dutch Research Council (NWO: Vici grant 016.140.657) to M.J. Smit; an NWO–Amsterdam Institute for Molecules, Medicines, and Systems STAR Graduate Program grant (022.005.031) and a Cancer Center Amsterdam travel grant to M.P. Bebelman; A European Molecular Biology Organization grant (EMBO ALTF 1383-2014) and a Fondation ARC pour la Recherch{\'e} sur le Cancer fellowship (PJA 20161204808) to F.J. Verweij; and a Fondation pour la Recherche M{\'e}dicale grant (AJE20160635884) and an Institut National Du Cancer grant (INCA N°2019-125 PLBIO19-059) to F.J. Verweij. and G. Van Niel. Conceptualization, M.P.B. D.M.P. M.S. and M.J.S.; Formal Analysis: M.P.B. I.M.S. and N.D.B.; Investigation: M.P.B. I.M.S. N.D.B. J.R.v.S. and J.P.B.; Writing – Original Draft: M.P.B.; Writing – Review & Editing: M.P.B. N.D.B. J.R.v.S. C.C. J.P.B. D.M.P. M.S. and M.J.S.; Visualization: M.P.B.; Supervision: C.C. F.J.V. M.S. D.M.P. and M.J.S.; Funding Acquisition: M.P.B. F.J.V. G.v.N. and M.J.S. M.J.S. is inventor of patent WO2019151865A1 describing VUN100. Funding Information: This work was funded by a grant from the Dutch Research Council (NWO: Vici grant 016.140.657 ) to M.J. Smit; an NWO–Amsterdam Institute for Molecules, Medicines, and Systems STAR Graduate Program grant ( 022.005.031 ) and a Cancer Center Amsterdam travel grant to M.P. Bebelman; A European Molecular Biology Organization grant ( EMBO ALTF 1383-2014 ) and a Fondation ARC pour la Recherch{\'e} sur le Cancer fellowship (PJA 20161204808 ) to F.J. Verweij; and a Fondation pour la Recherche M{\'e}dicale grant ( AJE20160635884 ) and an Institut National Du Cancer grant (INCA N°2019-125 PLBIO19-059 ) to F.J. Verweij. and G. Van Niel. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

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doi = "10.1016/j.isci.2023.107412",

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T1 - Exosomal release of the virus-encoded chemokine receptor US28 contributes to chemokine scavenging

AU - Bebelman, Maarten P.

AU - Setiawan, Irfan M.

AU - Bergkamp, Nick D.

AU - van Senten, Jeffrey R.

AU - Crudden, Caitrin

AU - Bebelman, Jan Paul M.

AU - Verweij, Frederik J.

AU - van Niel, Guillaume

AU - Siderius, Marco

AU - Pegtel, D. Michiel

AU - Smit, Martine J.

N1 - Funding Information: We thank R.J. Stanton (Cardiff University, Cardiff, UK) for sharing the HCMV Merlin BAC and HFFF-Tet cells. We greatly acknowledge the AO|2M microscopy core platform of VU University Medical Center Amsterdam and the Center for Neurogenetics and Cognitive Research imaging platform of the Neuroscience Campus Amsterdam for imaging support. We greatly acknowledge the Cell and Tissue Imaging Center (PICT-IBiSA) #Institut Curie for electron microscopy. This work was funded by a grant from the Dutch Research Council (NWO: Vici grant 016.140.657) to M.J. Smit; an NWO–Amsterdam Institute for Molecules, Medicines, and Systems STAR Graduate Program grant (022.005.031) and a Cancer Center Amsterdam travel grant to M.P. Bebelman; A European Molecular Biology Organization grant (EMBO ALTF 1383-2014) and a Fondation ARC pour la Recherché sur le Cancer fellowship (PJA 20161204808) to F.J. Verweij; and a Fondation pour la Recherche Médicale grant (AJE20160635884) and an Institut National Du Cancer grant (INCA N°2019-125 PLBIO19-059) to F.J. Verweij. and G. Van Niel. Conceptualization, M.P.B. D.M.P. M.S. and M.J.S.; Formal Analysis: M.P.B. I.M.S. and N.D.B.; Investigation: M.P.B. I.M.S. N.D.B. J.R.v.S. and J.P.B.; Writing – Original Draft: M.P.B.; Writing – Review & Editing: M.P.B. N.D.B. J.R.v.S. C.C. J.P.B. D.M.P. M.S. and M.J.S.; Visualization: M.P.B.; Supervision: C.C. F.J.V. M.S. D.M.P. and M.J.S.; Funding Acquisition: M.P.B. F.J.V. G.v.N. and M.J.S. M.J.S. is inventor of patent WO2019151865A1 describing VUN100. Funding Information: This work was funded by a grant from the Dutch Research Council (NWO: Vici grant 016.140.657 ) to M.J. Smit; an NWO–Amsterdam Institute for Molecules, Medicines, and Systems STAR Graduate Program grant ( 022.005.031 ) and a Cancer Center Amsterdam travel grant to M.P. Bebelman; A European Molecular Biology Organization grant ( EMBO ALTF 1383-2014 ) and a Fondation ARC pour la Recherché sur le Cancer fellowship (PJA 20161204808 ) to F.J. Verweij; and a Fondation pour la Recherche Médicale grant ( AJE20160635884 ) and an Institut National Du Cancer grant (INCA N°2019-125 PLBIO19-059 ) to F.J. Verweij. and G. Van Niel. Publisher Copyright: © 2023 The Author(s)

PY - 2023/8/18

Y1 - 2023/8/18

N2 - The human cytomegalovirus (HCMV)-encoded chemokine receptor US28 contributes to various aspects of the viral life cycle and promotes immune evasion by scavenging chemokines from the microenvironment of HCMV-infected cells. In contrast to the plasma membrane localization of most human chemokine receptors, US28 has a predominant intracellular localization. In this study, we used immunofluorescence and electron microscopy to determine the localization of US28 upon exogenous expression, as well as in HCMV-infected cells. We observed that US28 localizes to late endosomal compartments called multivesicular bodies (MVBs), where it is sorted in intraluminal vesicles. Live-cell total internal reflection fluorescence (TIRF) microscopy revealed that US28-containing MVBs can fuse with the plasma membrane, resulting in the secretion of US28 on exosomes. Exosomal US28 binds the chemokines CX3CL1 and CCL5, and US28-containing exosomes inhibited the CX3CL1-CX3CR1 signaling axis. These findings suggest that exosomal release of US28 contributes to chemokine scavenging and immune evasion by HCMV.

AB - The human cytomegalovirus (HCMV)-encoded chemokine receptor US28 contributes to various aspects of the viral life cycle and promotes immune evasion by scavenging chemokines from the microenvironment of HCMV-infected cells. In contrast to the plasma membrane localization of most human chemokine receptors, US28 has a predominant intracellular localization. In this study, we used immunofluorescence and electron microscopy to determine the localization of US28 upon exogenous expression, as well as in HCMV-infected cells. We observed that US28 localizes to late endosomal compartments called multivesicular bodies (MVBs), where it is sorted in intraluminal vesicles. Live-cell total internal reflection fluorescence (TIRF) microscopy revealed that US28-containing MVBs can fuse with the plasma membrane, resulting in the secretion of US28 on exosomes. Exosomal US28 binds the chemokines CX3CL1 and CCL5, and US28-containing exosomes inhibited the CX3CL1-CX3CR1 signaling axis. These findings suggest that exosomal release of US28 contributes to chemokine scavenging and immune evasion by HCMV.

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KW - Immunology

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Exosomal release of the virus-encoded chemokine receptor US28 contributes to chemokine scavenging

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