Wouter T Lollinga1, Raymond H de Wit, Afsar Rahbar, Gwenda F Vasse, Belghis Davoudi, Arjan Diepstra, Annelies Riezebos-Brilman, Martin C Harmsen, Jan-Luuk Hillebrands, Cecilia Söderberg-Naucler, Willem J van Son, Martine J Smit, Jan-Stephan Sanders, Jacob van den Born. 1. 1 Division of Nephrology, Department of Internal Medicine, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands. 2 Department of Chemistry and Pharmaceutical Sciences, Division of Medicinal Chemistry, Vrije Universiteit, Amsterdam, the Netherlands. 3 Department of Medicine, Center for Molecular Medicine, Unit for Microbial Pathogenesis, Karolinska Institutet, Solna, Stockholm, Sweden. 4 Division of Pathology, Department of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands. 5 Division of Clinical Virology, Department of Medical Microbiology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands. 6 Division of Medical Biology, Department of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
Abstract
BACKGROUND: Renal transplantation is the preferred treatment for patients with end-stage renal disease. Human cytomegalovirus (HCMV) activation is associated with decreased renal graft function and survival. Human cytomegalovirus encodes several immune modulatory proteins, including the G protein-coupled receptor US28, which scavenges human chemokines and modulates intracellular signaling. METHODS: Our aim was to identify the expression and localization of US28 in renal allograft biopsies by immunohistochemistry and determine its role in viral spreading in vitro. RESULTS: Immunohistochemistry revealed US28 in 31 of 34 renal transplant biopsies from HCMV-seropositive donors. Expression was independent of HCMV viremia or IgG serostatus. US28 was predominantly expressed in the cytoplasm of vascular smooth muscle cells (VSMCs) and tubular epithelial cells, with a median positivity of 20% and 40%, respectively. Also, US28-positive cells were present within arterial neointima. In contrast to US28, HCMV-encoded immediate early antigen was detected in less than 5% of VSMCs, tubular epithelial cells, interstitial endothelium, interstitial inflammatory infiltrates, and glomerular cells.Primary VSMCs were infected with green fluorescent protein-tagged wild type or US28-deficient HCMV. The viral spreading of US28-deficient HCMV, via culture medium or cell-to-cell transmission, was significantly impeded as shown by green fluorescent protein (ie, infected) cell quantification and quantitative real-time polymerase chain reaction. Additionally, the number and size of foci was smaller. CONCLUSIONS: In summary, HCMV-encoded US28 was detected in renal allografts from HCMV-positive donors independent of viremia and serostatus. Also, US28 facilitates HCMV spreading in VSMCs in vitro. Because the vasculature is affected in chronic renal transplant dysfunction, US28 may provide a potential target for therapeutic intervention.
BACKGROUND: Renal transplantation is the preferred treatment for patients with end-stage renal disease. Human cytomegalovirus (HCMV) activation is associated with decreased renal graft function and survival. Human cytomegalovirus encodes several immune modulatory proteins, including the G protein-coupled receptor US28, which scavenges human chemokines and modulates intracellular signaling. METHODS: Our aim was to identify the expression and localization of US28 in renal allograft biopsies by immunohistochemistry and determine its role in viral spreading in vitro. RESULTS: Immunohistochemistry revealed US28 in 31 of 34 renal transplant biopsies from HCMV-seropositive donors. Expression was independent of HCMV viremia or IgG serostatus. US28 was predominantly expressed in the cytoplasm of vascular smooth muscle cells (VSMCs) and tubular epithelial cells, with a median positivity of 20% and 40%, respectively. Also, US28-positive cells were present within arterial neointima. In contrast to US28, HCMV-encoded immediate early antigen was detected in less than 5% of VSMCs, tubular epithelial cells, interstitial endothelium, interstitial inflammatory infiltrates, and glomerular cells.Primary VSMCs were infected with green fluorescent protein-tagged wild type or US28-deficient HCMV. The viral spreading of US28-deficient HCMV, via culture medium or cell-to-cell transmission, was significantly impeded as shown by green fluorescent protein (ie, infected) cell quantification and quantitative real-time polymerase chain reaction. Additionally, the number and size of foci was smaller. CONCLUSIONS: In summary, HCMV-encoded US28 was detected in renal allografts from HCMV-positive donors independent of viremia and serostatus. Also, US28 facilitates HCMV spreading in VSMCs in vitro. Because the vasculature is affected in chronic renal transplant dysfunction, US28 may provide a potential target for therapeutic intervention.
Authors: Mette M Rosenkilde; Naotaka Tsutsumi; Julius M Knerr; Dagmar F Kildedal; K Christopher Garcia Journal: Annu Rev Virol Date: 2022-06-07 Impact factor: 14.263
Authors: Raimond Heukers; Tian Shu Fan; Raymond H de Wit; Jeffrey R van Senten; Timo W M De Groof; Maarten P Bebelman; Tonny Lagerweij; Joao Vieira; Sabrina M de Munnik; Laura Smits-de Vries; Jody van Offenbeek; Afsar Rahbar; Diane van Hoorick; Cecilia Söderberg-Naucler; Thomas Würdinger; Rob Leurs; Marco Siderius; Henry F Vischer; Martine J Smit Journal: Oncogene Date: 2018-04-30 Impact factor: 9.867
Authors: Jeffrey R van Senten; Maarten P Bebelman; Puck van Gasselt; Nick D Bergkamp; Jelle van den Bor; Marco Siderius; Martine J Smit Journal: Viruses Date: 2020-05-30 Impact factor: 5.048