Attenuation of chemokine receptor function and surface expression as an immunomodulatory strategy employed by human cytomegalovirus is linked to vGPCR US28Reportar como inadecuado

Attenuation of chemokine receptor function and surface expression as an immunomodulatory strategy employed by human cytomegalovirus is linked to vGPCR US28 - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Cell Communication and Signaling

, 14:31

First Online: 12 December 2016Received: 27 June 2016Accepted: 02 December 2016


BackgroundSome herpesviruses like human cytomegalovirus HCMV encode viral G protein-coupled receptors that cause reprogramming of cell signaling to facilitate dissemination of the virus, prevent immune surveillance and establish life-long latency. Human GPCRs are known to function in complex signaling networks involving direct physical interactions as well as indirect crosstalk of orthogonal signaling networks. The human chemokine receptor CXCR4 is expressed on hematopoietic stem cells, leukocytes, endothelial and epithelial cells, which are infected by HCMV or display reservoirs of latency.

ResultsWe investigated the potential heteromerization of US28 with CXCR4 as well as the influence of US28 on CXCR4 signaling. Using Bioluminescence Resonance Energy Transfer and luciferase-complementation based methods we show that US28 expression exhibits negative effects on CXCR4 signaling and constitutive surface expression in HEK293T cells. Furthermore, we demonstrate that this effect is not mediated by receptor heteromerization but via signaling crosstalk. Additionally, we show that in HCMV, strain TB40E, infected HUVEC the surface expression of CXCR4 is strongly downregulated, whereas in TB40E-delUS28 infected cells, CXCR4 surface expression is not altered in particular at late time points of infection.

ConclusionsWe show that the vGPCR US28 is leading to severely disturbed signaling and surface expression of the chemokine receptor CXCR4 thereby representing an effective mechanism used by vGPCRs to reprogram host cell signaling. In contrast to other studies, we demonstrate that these effects are not mediated via heteromerization.

KeywordsViral G protein-coupled receptor US28 Chemokine receptor CXCR4 Constitutive activity Bioluminescence resonance energy transfer Bioluminescence complementation Signaling crosstalk AbbreviationsACAdenylate cyclase

BACBacterial artificial chromosome

BiLCBioluminescence complementation

BRETBioluminescence resonance energy transfer

EBVEpstein Barr virus

ECEndothelial cells

ELISAEnzyme-linked immunosorbent assay

Elucemerald luciferase

FACSFluorescence-activated cell sorting

FBSFetal bovine serum

Flucfirefly luciferase

FRAPFluorescence recovery after photobleaching

FRETFluorescence resonance energy transfer

HCMVHuman cytomegalovirus

HIVHuman immunodeficiency virus

HSPCsHematopoietic stem and progenitor cells

hpiHours postinfection

HUVECHuman umbilical vein endothelial cells

IC50half maximal inhibitory concentration

KSHVKaposi-s sarcoma-associated herpesvirus

MCP-1Monocyte chemotactic protein-1

MOImultiplicity of infection

NTS1Neurotensin receptor type 1

RANTESRegulated on activation, normal T-cell expressed and secreted

Rluc8Renilla reniformis luciferase 8

SDF1αStromal cell-derived factor-1α

vGPCRsviral G protein-coupled receptors

Electronic supplementary materialThe online version of this article doi:10.1186-s12964-016-0154-x contains supplementary material, which is available to authorized users.

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Autor: Theresa Frank - Anna Reichel - Olav Larsen - Anne-Charlotte Stilp - Mette M. Rosenkilde - Thomas Stamminger - Takeaki Ozaw


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