Tail Class I Molecules Through Their Cytoplasmic Surface Display of a Wide Range of HLA EBV BILF1 Evolved to Downregulate Cell
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EBV BILF1 Evolved To Downregulate Cell Surface Display of a Wide Range of HLA Class I Molecules through Their Cytoplasmic Tail This information is current as of October 1, 2021. Bryan D. Griffin, Anna M. Gram, Arend Mulder, Daphne Van Leeuwen, Frans H. J. Claas, Fred Wang, Maaike E. Ressing and Emmanuel Wiertz J Immunol 2013; 190:1672-1684; Prepublished online 11 January 2013; Downloaded from doi: 10.4049/jimmunol.1102462 http://www.jimmunol.org/content/190/4/1672 Supplementary http://www.jimmunol.org/content/suppl/2013/01/14/jimmunol.110246 http://www.jimmunol.org/ Material 2.DC1 References This article cites 63 articles, 29 of which you can access for free at: http://www.jimmunol.org/content/190/4/1672.full#ref-list-1 Why The JI? Submit online. by guest on October 1, 2021 • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology EBV BILF1 Evolved To Downregulate Cell Surface Display of a Wide Range of HLA Class I Molecules through Their Cytoplasmic Tail Bryan D. Griffin,*,†,1 Anna M. Gram,*,1 Arend Mulder,‡ Daphne Van Leeuwen,† Frans H. J. Claas,‡ Fred Wang,x Maaike E. Ressing,*,†,2 and Emmanuel Wiertz*,†,2 Coevolution of herpesviruses and their hosts has driven the development of both host antiviral mechanisms to detect and eliminate infected cells and viral ploys to escape immune surveillance. Among the immune-evasion strategies used by the lymphocryptovirus (g1-herpesvirus) EBV is the downregulation of surface HLA class I expression by the virally encoded G protein–coupled receptor BILF1, thereby impeding presentation of viral Ags and cytotoxic T cell recognition of the infected cell. In this study, we show EBV BILF1 to be expressed early in the viral lytic cycle. BILF1 targets a broad range of HLA class I molecules, including multiple HLA- A and -B types and HLA-E. In contrast, HLA-C was only marginally affected. We advance the mechanistic understanding of the Downloaded from process by showing that the cytoplasmic C-terminal tail of EBV BILF1 is required for reducing surface HLA class I expression. Susceptibility to BILF1-mediated downregulation, in turn, is conferred by specific residues in the intracellular tail of the HLA class I H chain. Finally, we explore the evolution of BILF1 within the lymphocryptovirus genus. Although the homolog of BILF1 encoded by the lymphocryptovirus infecting Old World rhesus primates shares the ability of EBV to downregulate cell surface HLA class I expression, this function is not possessed by New World marmoset lymphocryptovirus BILF1. Therefore, this study furthers our knowledge of the evolution of immunoevasive functions by the lymphocryptovirus genus of herpesviruses. The Journal of Immu- http://www.jimmunol.org/ nology, 2013, 190: 1672–1684. ymphocryptoviruses (LCVs) are a genus of the g-her- individuals (5). This is partly due to the ability of EBV, like all pesvirus subfamily whose members are only found in herpesviruses, to enter a state of latency in which protein ex- L primates (1). The LCV targeting humans, EBV, is carried pression is minimized, thereby limiting viral Ag display by the by .90% of adults worldwide (2, 3). Although infection is usually infected cell. Yet, for EBV to spread to a new host, it must pro- asymptomatic, primary encounter with the virus can present as duce infectious virions by entering the replicative, or lytic, phase of its life cycle. In this phase, .80 EBV-encoded proteins are infectious mononucleosis. EBV infection is also strongly associ- by guest on October 1, 2021 ated with tumors of lymphoid and epithelial origin, reflecting the expressed in a temporal cascade, with initial production of im- tropism of the virus for B cells and epithelial cells and its potential mediate early (IE) transactivators triggering induction of early for oncogenic transformation (4). genes, including enzymes required for viral replication (2). This Despite the activation of a robust host T cell response upon is followed by expression of late genes encoding virion structural primary infection and the capacity for a memory T cell response components. Thus, the lytic phase creates ample viral Ags for thereafter, the virus persists for life even in immunocompetent proteasomal processing and HLA class I cell surface presentation, which can then recruit memory CD8+ T cells capable of elimi- nating the infected cell. However, millions of years of coevolution *Department of Medical Microbiology, University Medical Center Utrecht, 3584 CX with their hosts have seen herpesviruses acquire active immune- Utrecht, The Netherlands; †Department of Medical Microbiology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands; ‡Department of Immunohaema- evasion mechanisms to thwart this host response and permit suf- tology and Blood Transfusion, Leiden University Medical Center, 2300 RC Leiden, ficient time for the lytically infected cell to generate new virus The Netherlands; and xDepartment of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115 particles (6–8). In the case of EBV, several such immune-evasion strategies 1B.D.G. and A.M.G. contributed equally to this work. target the HLA class I Ag-processing and -presentation pathways 2M.E.R. and E.W. contributed equally to this work. (9). The EBV host shutoff protein BGLF5 degrades mRNA, thus Received for publication September 6, 2011. Accepted for publication December 11, 2012. obstructing synthesis of new HLA class I molecules (10, 11). Meanwhile, BNLF2a blocks entry of proteasome-generated pep- This work was supported by Dutch Cancer Foundation Grant UL 2005-3259 (to B.D.G. and E.W.), Netherlands Organisation of Scientific Research Vidi Grant 917.76.330 (to tides into the endoplasmic reticulum (ER) and subsequent loading M.E.R.), and National Institutes of Health Grant CA068051 (to F.W.). onto HLA class I molecules by inhibiting the heterodimeric TAP Address correspondence and reprint requests to Prof. Emmanuel Wiertz, Department complex (12, 13). TAP transport is also hindered by the viral che- of Medical Microbiology G04.614, University Medical Center Utrecht, P.O. Box mokine homolog, vIL-10, which reduces expression of the TAP1 85500, 3508 GA Utrecht, The Netherlands. E-mail address: [email protected] subunit (14). Another manner in which EBV can downregulate cell The online version of this article contains supplemental material. surface HLA class I expression and inhibit T cell recognition of Abbreviations used in this article: ER, endoplasmic reticulum; GPCR, G protein– coupled receptor; HC, H chain; IE, immediate early; IRES, internal ribosome entry infected cells is through BILF1, a viral G protein–coupled receptor site; LCV, lymphocryptovirus; NGFR, nerve growth factor receptor; PAA, phospho- (vGPCR) (15, 16). noacetic acid; trNGFR, truncated nerve growth factor receptor; vGPCR, viral G Several poxviruses and herpesviruses encode G protein–coupled protein–coupled receptor; wt, wild type. receptors (GPCRs) that were most likely pirated from their host Copyright Ó 2013 by The American Association of Immunologists, Inc. 0022-1767/13/$16.00 by retrotransposition (17–19). vGPCRs serve many functions, www.jimmunol.org/cgi/doi/10.4049/jimmunol.1102462 The Journal of Immunology 1673 including the scavenging of host chemokines (20), cell-to-cell ad- R.J. Lebbink, University Medical Center Utrecht, The Netherlands). The hesion (21), and the reprogramming of intracellular signaling net- pSicoR-EGFP backbone vector (Addgene) was altered by removing the U6 works to promote efficient viral replication (22). BILF1 is expressed promoter and replacing the CMV-EGFP cassette with the human EF1A promoter driving expression of the Zeocin resistance gene and the gene of during the EBV lytic cycle and was first identified as a potential interest. Zeocin and HLA-B8 genes were fused together by a self-cleaving vGPCR by the presence of seven membrane-spanning domains and 2A peptide derived from the porcine teschovirus-1 (P2A). The P2A peptide (limited) homology with known herpesviral GPCRs (23–25). Al- allows expression of both proteins from a single transcript. HLA-B8wt was though it displays both structural and functional similarities to amplified by PCR from the bidirectional lentiviral vector mentioned above; the cytoplasmic tail mutants were generated by using reverse primers that chemokine receptors, EBV BILF1 modulates intracellular signaling encoded the mutations. pathways constitutively as an orphan receptor (24, 25). Restriction digests and sequence analysis verified the integrity of all gene Initial immune-evasion ability was ascribed to EBV BILF1 sequences. because it reduced phosphorylation of the dsRNA-dependent Cell lines protein kinase R (24). It was also found to heterodimerize with + human chemokine receptors (26). In the case of CXCR4, this The AKBM cell line is an EBV Burkitt’s lymphoma B cell line (Akata) results in impairment of ligand-induced receptor signaling (27). stably transfected with the pHEBO-BMRF1p-rCD2-GFP reporter plasmid (30). AKBM cells were maintained in RPMI 1640 medium (Invitrogen) Finally, EBV BILF1 decreases cell-surface levels of HLA class I supplemented with 10% FBS (PAA Laboratories, Pasching, Austria), 2 (15). This can be achieved by BILF1-mediated acceleration mM L-glutamine, 100 U/ml penicillin, 100 mg/ml streptomycin, and 0.3 of endocytosis and subsequent lysosomal degradation of HLA class mg/ml hygromycin B.