261 results on '"Alan B. Rickinson"'
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2. Data from A Recombinant Modified Vaccinia Ankara Vaccine Encoding Epstein–Barr Virus (EBV) Target Antigens: A Phase I Trial in UK Patients with EBV-Positive Cancer
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Neil M. Steven, Alan B. Rickinson, Anthony T. C. Chan, Edwin P. Hui, Steve Wilson, Andrew Hartley, Lesley McGuigan, Ceri Edwards, Claudia Roberts, Jen Matthews, Manjit Tanday, David A. Price, Kristin Ladell, James Turner, Lip Wai Lee, Kevin Harrington, Hui Jia, and Graham S. Taylor
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Purpose: Epstein–Barr virus (EBV) is associated with several cancers in which the tumor cells express EBV antigens EBNA1 and LMP2. A therapeutic vaccine comprising a recombinant vaccinia virus, MVA-EL, was designed to boost immunity to these tumor antigens. A phase I trial was conducted to demonstrate the safety and immunogenicity of MVA-EL across a range of doses.Experimental Design: Sixteen patients in the United Kingdom (UK) with EBV-positive nasopharyngeal carcinoma (NPC) received three intradermal vaccinations of MVA-EL at 3-weekly intervals at dose levels between 5 × 107 and 5 × 108 plaque-forming units (pfu). Blood samples were taken at screening, after each vaccine cycle, and during the post-vaccination period. T-cell responses were measured using IFNγ ELISpot assays with overlapping EBNA1/LMP2 peptide mixes or HLA-matched epitope peptides. Polychromatic flow cytometry was used to characterize functionally responsive T-cell populations.Results: Vaccination was generally well tolerated. Immunity increased after vaccination to at least one antigen in 8 of 14 patients (7/14, EBNA1; 6/14, LMP2), including recognition of epitopes that vary between EBV strains associated with different ethnic groups. Immunophenotypic analysis revealed that vaccination induced differentiation and functional diversification of responsive T-cell populations specific for EBNA1 and LMP2 within the CD4 and CD8 compartments, respectively.Conclusions: MVA-EL is safe and immunogenic across diverse ethnicities and thus suitable for use in trials against different EBV-positive cancers globally as well as in South-East Asia where NPC is most common. The highest dose (5 × 108 pfu) is recommended for investigation in current phase IB and II trials. Clin Cancer Res; 20(19); 5009–22. ©2014 AACR.
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- 2023
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3. Supplementary Methods, Figures 1 - 2, Tables 1 - 3 from Phase I Trial of Recombinant Modified Vaccinia Ankara Encoding Epstein–Barr Viral Tumor Antigens in Nasopharyngeal Carcinoma Patients
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Anthony T.C. Chan, Neil M. Steven, Alan B. Rickinson, Deborah D. Stocken, Ceri Edwards, Benjamin F. Johnson, Steven Wilson, Wai-Lap Wong, Rosalie Ho, Stephen L. Chan, Brigette B.Y. Ma, Hui Jia, Graham S. Taylor, and Edwin P. Hui
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PDF file - 269K, Additional detail of methodology, adverse events, epitope peptides and epitope-specific T cell responses. Also includes all results obtained using ELIspot assays with overlapping peptides and analysis of regulatory T cells in patients.
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- 2023
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4. Proteome-wide analysis of CD8+ T cell responses to EBV reveals differences between primary and persistent infection.
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Calum Forrest, Andrew D Hislop, Alan B Rickinson, and Jianmin Zuo
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Human herpesviruses are antigenically rich agents that induce strong CD8+T cell responses in primary infection yet persist for life, continually challenging T cell memory through recurrent lytic replication and potentially influencing the spectrum of antigen-specific responses. Here we describe the first lytic proteome-wide analysis of CD8+ T cell responses to a gamma1-herpesvirus, Epstein-Barr virus (EBV), and the first such proteome-wide analysis of primary versus memory CD8+ T cell responses to any human herpesvirus. Primary effector preparations were generated directly from activated CD8+ T cells in the blood of infectious mononucleosis (IM) patients by in vitro mitogenic expansion. For memory preparations, EBV-specific cells in the blood of long-term virus carriers were first re-stimulated in vitro by autologous dendritic cells loaded with a lysate of lytically-infected cells, then expanded as for IM cells. Preparations from 7 donors of each type were screened against each of 70 EBV lytic cycle proteins in combination with the donor's individual HLA class I alleles. Multiple reactivities against immediate early (IE), early (E) and late (L) lytic cycle proteins, including many hitherto unrecognised targets, were detected in both contexts. Interestingly however, the two donor cohorts showed a different balance between IE, E and L reactivities. Primary responses targeted IE and a small group of E proteins preferentially, seemingly in line with their better presentation on the infected cell surface before later-expressed viral evasins take full hold. By contrast, target choice equilibrates in virus carriage with responses to key IE and E antigens still present but with responses to a select subset of L proteins now often prominent. We infer that, for EBV at least, long-term virus carriage with its low level virus replication and lytic antigen release is associated with a re-shaping of the virus-specific response.
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- 2018
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5. Compartmentalization of Total and Virus-Specific Tissue-Resident Memory CD8+ T Cells in Human Lymphoid Organs.
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Heng Giap Woon, Asolina Braun, Jane Li, Corey Smith, Jarem Edwards, Frederic Sierro, Carl G Feng, Rajiv Khanna, Michael Elliot, Andrew Bell, Andrew D Hislop, Stuart G Tangye, Alan B Rickinson, Thomas Gebhardt, Warwick J Britton, and Umaimainthan Palendira
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Disruption of T cell memory during severe immune suppression results in reactivation of chronic viral infections, such as Epstein Barr virus (EBV) and Cytomegalovirus (CMV). How different subsets of memory T cells contribute to the protective immunity against these viruses remains poorly defined. In this study we examined the compartmentalization of virus-specific, tissue resident memory CD8+ T cells in human lymphoid organs. This revealed two distinct populations of memory CD8+ T cells, that were CD69+CD103+ and CD69+CD103-, and were retained within the spleen and tonsils in the absence of recent T cell stimulation. These two types of memory cells were distinct not only in their phenotype and transcriptional profile, but also in their anatomical localization within tonsils and spleen. The EBV-specific, but not CMV-specific, CD8+ memory T cells preferentially accumulated in the tonsils and acquired a phenotype that ensured their retention at the epithelial sites where EBV replicates. In vitro studies revealed that the cytokine IL-15 can potentiate the retention of circulating effector memory CD8+ T cells by down-regulating the expression of sphingosine-1-phosphate receptor, required for T cell exit from tissues, and its transcriptional activator, Kruppel-like factor 2 (KLF2). Within the tonsils the expression of IL-15 was detected in regions where CD8+ T cells localized, further supporting a role for this cytokine in T cell retention. Together this study provides evidence for the compartmentalization of distinct types of resident memory T cells that could contribute to the long-term protection against persisting viral infections.
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- 2016
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6. Early T Cell Recognition of B Cells following Epstein-Barr Virus Infection: Identifying Potential Targets for Prophylactic Vaccination.
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Jill M Brooks, Heather M Long, Rose J Tierney, Claire Shannon-Lowe, Alison M Leese, Martin Fitzpatrick, Graham S Taylor, and Alan B Rickinson
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Epstein-Barr virus, a B-lymphotropic herpesvirus, is the cause of infectious mononucleosis, has strong aetiologic links with several malignancies and has been implicated in certain autoimmune diseases. Efforts to develop a prophylactic vaccine to prevent or reduce EBV-associated disease have, to date, focused on the induction of neutralising antibody responses. However, such vaccines might be further improved by inducing T cell responses capable of recognising and killing recently-infected B cells. In that context, EBNA2, EBNA-LP and BHRF1 are the first viral antigens expressed during the initial stage of B cell growth transformation, yet have been poorly characterised as CD8+ T cell targets. Here we describe CD8+ T cell responses against each of these three "first wave" proteins, identifying target epitopes and HLA restricting alleles. While EBNA-LP and BHRF1 each contained one strong CD8 epitope, epitopes within EBNA2 induced immunodominant responses through several less common HLA class I alleles (e.g. B*3801 and B*5501), as well as subdominant responses through common class I alleles (e.g. B7 and C*0304). Importantly, such EBNA2-specific CD8+ T cells recognised B cells within the first day post-infection, prior to CD8+ T cells against well-characterised latent target antigens such as EBNA3B or LMP2, and effectively inhibited outgrowth of EBV-transformed B cell lines. We infer that "first wave" antigens of the growth-transforming infection, especially EBNA2, constitute potential CD8+ T cell immunogens for inclusion in prophylactic EBV vaccine design.
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- 2016
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7. Early virological and immunological events in asymptomatic Epstein-Barr virus infection in African children.
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Shamanthi Jayasooriya, Thushan I de Silva, Jainaba Njie-jobe, Chilel Sanyang, Alison M Leese, Andrew I Bell, Karen A McAulay, Peng Yanchun, Heather M Long, Tao Dong, Hilton C Whittle, Alan B Rickinson, Sarah L Rowland-Jones, Andrew D Hislop, and Katie L Flanagan
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Epstein-Barr virus (EBV) infection often occurs in early childhood and is asymptomatic. However, if delayed until adolescence, primary infection may manifest as acute infectious mononucleosis (AIM), a febrile illness characterised by global CD8+ T-cell lymphocytosis, much of it reflecting a huge expansion of activated EBV-specific CD8+ T-cells. While the events of AIM have been intensely studied, little is known about how these relate to asymptomatic primary infection. Here Gambian children (14-18 months old, an age at which many acquire the virus) were followed for the ensuing six months, monitoring circulating EBV loads, antibody status against virus capsid antigen (VCA) and both total and virus-specific CD8+ T-cell numbers. Many children were IgG anti-VCA-positive and, though no longer IgM-positive, still retained high virus loads comparable to AIM patients and had detectable EBV-specific T-cells, some still expressing activation markers. Virus loads and the frequency/activation status of specific T-cells decreased over time, consistent with resolution of a relatively recent primary infection. Six children with similarly high EBV loads were IgM anti-VCA-positive, indicating very recent infection. In three of these donors with HLA types allowing MHC-tetramer analysis, highly activated EBV-specific T-cells were detectable in the blood with one individual epitope response reaching 15% of all CD8+ T-cells. That response was culled and the cells lost activation markers over time, just as seen in AIM. However, unlike AIM, these events occurred without marked expansion of total CD8+ numbers. Thus asymptomatic EBV infection in children elicits a virus-specific CD8+ T-cell response that can control the infection without over-expansion; conversely, in AIM it appears the CD8 over-expansion, rather than virus load per se, is the cause of disease symptoms.
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- 2015
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8. Epstein–Barr virus
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Alan B. Rickinson and M.A. Epstein
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hemic and lymphatic diseases ,viruses - Abstract
Epstein–Barr virus is a human herpesvirus with a linear double-stranded DNA genome that is carried asymptomatically by most people. Symptomless primary infection is usual in childhood, establishing a lifelong carrier state where the virus persists as a latent infection of circulating B cells. The virus replicates recurrently in oropharyngeal epithelial cells, with consequent shedding of virus in saliva transmitting infection. Controversially, Epstein–Barr virus has been linked with certain autoimmune diseases. In particular, there is strong serologic and epidemiologic evidence to suggest that previous exposure to Epstein–Barr virus markedly increases the risk of developing multiple sclerosis. Although the Epstein–Barr virus/multiple sclerosis connection is receiving much attention, the mechanism that might underpin such an association remains uncertain.
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- 2020
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9. Epstein-Barr virus infection of naïve B cells in vitro frequently selects clones with mutated immunoglobulin genotypes: implications for virus biology.
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Emily Heath, Noelia Begue-Pastor, Sridhar Chaganti, Debbie Croom-Carter, Claire Shannon-Lowe, Dieter Kube, Regina Feederle, Henri-Jacques Delecluse, Alan B Rickinson, and Andrew I Bell
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Epstein-Barr virus (EBV), a lymphomagenic human herpesvirus, colonises the host through polyclonal B cell-growth-transforming infections yet establishes persistence only in IgD⁺ CD27⁺ non-switched memory (NSM) and IgD⁻ CD27⁺ switched memory (SM) B cells, not in IgD⁺ CD27⁻ naïve (N) cells. How this selectivity is achieved remains poorly understood. Here we show that purified N, NSM and SM cell preparations are equally transformable in vitro to lymphoblastoid cells lines (LCLs) that, despite upregulating the activation-induced cytidine deaminase (AID) enzyme necessary for Ig isotype switching and Ig gene hypermutation, still retain the surface Ig phenotype of their parental cells. However, both N- and NSM-derived lines remain inducible to Ig isotype switching by surrogate T cell signals. More importantly, IgH gene analysis of N cell infections revealed two features quite distinct from parallel mitogen-activated cultures. Firstly, following 4 weeks of EBV-driven polyclonal proliferation, individual clonotypes then become increasingly dominant; secondly, in around 35% cases these clonotypes carry Ig gene mutations which both resemble AID products and, when analysed in prospectively-harvested cultures, appear to have arisen by sequence diversification in vitro. Thus EBV infection per se can drive at least some naïve B cells to acquire Ig memory genotypes; furthermore, such cells are often favoured during an LCL's evolution to monoclonality. Extrapolating to viral infections in vivo, these findings could help to explain how EBV-infected cells become restricted to memory B cell subsets and why EBV-driven lymphoproliferative lesions, in primary infection and/or immunocompromised settings, so frequently involve clones with memory genotypes.
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- 2012
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10. Molecular pathogenesis of EBV susceptibility in XLP as revealed by analysis of female carriers with heterozygous expression of SAP.
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Umaimainthan Palendira, Carol Low, Anna Chan, Andrew D Hislop, Edwin Ho, Tri Giang Phan, Elissa Deenick, Matthew C Cook, D Sean Riminton, Sharon Choo, Richard Loh, Frank Alvaro, Claire Booth, H Bobby Gaspar, Alessandro Moretta, Rajiv Khanna, Alan B Rickinson, and Stuart G Tangye
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Biology (General) ,QH301-705.5 - Abstract
X-linked lymphoproliferative disease (XLP) is a primary immunodeficiency caused by mutations in SH2D1A which encodes SAP. SAP functions in signalling pathways elicited by the SLAM family of leukocyte receptors. A defining feature of XLP is exquisite sensitivity to infection with EBV, a B-lymphotropic virus, but not other viruses. Although previous studies have identified defects in lymphocytes from XLP patients, the unique role of SAP in controlling EBV infection remains unresolved. We describe a novel approach to this question using female XLP carriers who, due to random X-inactivation, contain both SAP(+) and SAP(-) cells. This represents the human equivalent of a mixed bone marrow chimera in mice. While memory CD8(+) T cells specific for CMV and influenza were distributed across SAP(+) and SAP(-) populations, EBV-specific cells were exclusively SAP(+). The preferential recruitment of SAP(+) cells by EBV reflected the tropism of EBV for B cells, and the requirement for SAP expression in CD8(+) T cells for them to respond to Ag-presentation by B cells, but not other cell types. The inability of SAP(-) clones to respond to Ag-presenting B cells was overcome by blocking the SLAM receptors NTB-A and 2B4, while ectopic expression of NTB-A on fibroblasts inhibited cytotoxicity of SAP(-) CD8(+) T cells, thereby demonstrating that SLAM receptors acquire inhibitory function in the absence of SAP. The innovative XLP carrier model allowed us to unravel the mechanisms underlying the unique susceptibility of XLP patients to EBV infection in the absence of a relevant animal model. We found that this reflected the nature of the Ag-presenting cell, rather than EBV itself. Our data also identified a pathological signalling pathway that could be targeted to treat patients with severe EBV infection. This system may allow the study of other human diseases where heterozygous gene expression from random X-chromosome inactivation can be exploited.
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- 2011
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11. T cell detection of a B-cell tropic virus infection: newly-synthesised versus mature viral proteins as antigen sources for CD4 and CD8 epitope display.
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Laura K Mackay, Heather M Long, Jill M Brooks, Graham S Taylor, Carol S Leung, Adrienne Chen, Fred Wang, and Alan B Rickinson
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Viruses that naturally infect cells expressing both MHC I and MHC II molecules render themselves potentially visible to both CD8+ and CD4+ T cells through the de novo expression of viral antigens. Here we use one such pathogen, the B-lymphotropic Epstein-Barr virus (EBV), to examine the kinetics of these processes in the virally-infected cell, comparing newly synthesised polypeptides versus the mature protein pool as viral antigen sources for MHC I- and MHC II-restricted presentation. EBV-transformed B cell lines were established in which the expression of two cognate EBV antigens, EBNA1 and EBNA3B, could be induced and then completely suppressed by doxycycline-regulation. These cells were used as targets for CD8+ and CD4+ T cell clones to a range of EBNA1 and EBNA3B epitopes. For both antigens, when synthesis was induced, CD8 epitope display rose quickly to near maximum within 24 h, well before steady state levels of mature protein had been reached, whereas CD4 epitope presentation was delayed by 36-48 h and rose only slowly thereafter. When antigen expression was suppressed, despite the persistence of mature protein, CD8 epitope display fell rapidly at rates similar to that seen for the MHC I/epitope half-life in peptide pulse-chase experiments. By contrast, CD4 epitope display persisted for many days and, following peptide stripping, recovered well on cells in the absence of new antigen synthesis. We infer that, in virally-infected MHC I/II-positive cells, newly-synthesised polypeptides are the dominant source of antigen feeding the MHC I pathway, whereas the MHC II pathway is fed by the mature protein pool. Hence, newly-infected cells are rapidly visible only to the CD8 response; by contrast, latent infections, in which viral gene expression has been extinguished yet viral proteins persist, will remain visible to CD4+ T cells.
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- 2009
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12. Stage-specific inhibition of MHC class I presentation by the Epstein-Barr virus BNLF2a protein during virus lytic cycle.
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Nathan P Croft, Claire Shannon-Lowe, Andrew I Bell, Daniëlle Horst, Elisabeth Kremmer, Maaike E Ressing, Emmanuel J H J Wiertz, Jaap M Middeldorp, Martin Rowe, Alan B Rickinson, and Andrew D Hislop
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The gamma-herpesvirus Epstein-Barr virus (EBV) persists for life in infected individuals despite the presence of a strong immune response. During the lytic cycle of EBV many viral proteins are expressed, potentially allowing virally infected cells to be recognized and eliminated by CD8+ T cells. We have recently identified an immune evasion protein encoded by EBV, BNLF2a, which is expressed in early phase lytic replication and inhibits peptide- and ATP-binding functions of the transporter associated with antigen processing. Ectopic expression of BNLF2a causes decreased surface MHC class I expression and inhibits the presentation of indicator antigens to CD8+ T cells. Here we sought to examine the influence of BNLF2a when expressed naturally during EBV lytic replication. We generated a BNLF2a-deleted recombinant EBV (DeltaBNLF2a) and compared the ability of DeltaBNLF2a and wild-type EBV-transformed B cell lines to be recognized by CD8+ T cell clones specific for EBV-encoded immediate early, early and late lytic antigens. Epitopes derived from immediate early and early expressed proteins were better recognized when presented by DeltaBNLF2a transformed cells compared to wild-type virus transformants. However, recognition of late antigens by CD8+ T cells remained equally poor when presented by both wild-type and DeltaBNLF2a cell targets. Analysis of BNLF2a and target protein expression kinetics showed that although BNLF2a is expressed during early phase replication, it is expressed at a time when there is an upregulation of immediate early proteins and initiation of early protein synthesis. Interestingly, BNLF2a protein expression was found to be lost by late lytic cycle yet DeltaBNLF2a-transformed cells in late stage replication downregulated surface MHC class I to a similar extent as wild-type EBV-transformed cells. These data show that BNLF2a-mediated expression is stage-specific, affecting presentation of immediate early and early proteins, and that other evasion mechanisms operate later in the lytic cycle.
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- 2009
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13. An Epstein-Barr virus anti-apoptotic protein constitutively expressed in transformed cells and implicated in burkitt lymphomagenesis: the Wp/BHRF1 link.
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Gemma L Kelly, Heather M Long, Julianna Stylianou, Wendy A Thomas, Alison Leese, Andrew I Bell, Georg W Bornkamm, Josef Mautner, Alan B Rickinson, and Martin Rowe
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Two factors contribute to Burkitt lymphoma (BL) pathogenesis, a chromosomal translocation leading to c-myc oncogene deregulation and infection with Epstein-Barr virus (EBV). Although the virus has B cell growth-transforming ability, this may not relate to its role in BL since many of the transforming proteins are not expressed in the tumor. Mounting evidence supports an alternative role, whereby EBV counteracts the high apoptotic sensitivity inherent to the c-myc-driven growth program. In that regard, a subset of BLs carry virus mutants in a novel form of latent infection that provides unusually strong resistance to apoptosis. Uniquely, these virus mutants use Wp (a viral promoter normally activated early in B cell transformation) and express a broader-than-usual range of latent antigens. Here, using an inducible system to express the candidate antigens, we show that this marked apoptosis resistance is mediated not by one of the extended range of EBNAs seen in Wp-restricted latency but by Wp-driven expression of the viral bcl2 homologue, BHRF1, a protein usually associated with the virus lytic cycle. Interestingly, this Wp/BHRF1 connection is not confined to Wp-restricted BLs but appears integral to normal B cell transformation by EBV. We find that the BHRF1 gene expression recently reported in newly infected B cells is temporally linked to Wp activation and the presence of W/BHRF1-spliced transcripts. Furthermore, just as Wp activity is never completely eclipsed in in vitro-transformed lines, low-level BHRF1 transcripts remain detectable in these cells long-term. Most importantly, recognition by BHRF1-specific T cells confirms that such lines continue to express the protein independently of any lytic cycle entry. This work therefore provides the first evidence that BHRF1, the EBV bcl2 homologue, is constitutively expressed as a latent protein in growth-transformed cells in vitro and, in the context of Wp-restricted BL, may contribute to virus-associated lymphomagenesis in vivo.
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- 2009
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14. Transient reduction in IgA(+) and IgG(+) memory B cell numbers in young EBV-seropositive children: the Generation R Study
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Andrew I. Bell, Jacques J.M. van Dongen, Michelle A. E. Jansen, Menno C. van Zelm, Diana van den Heuvel, Alan B. Rickinson, Henriëtte A. Moll, Vincent W. V. Jaddoe, Immunology, Erasmus MC other, Pediatrics, and Epidemiology
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0301 basic medicine ,Immunology ,Cell ,chemical and pharmacologic phenomena ,Biology ,medicine.disease_cause ,Measles ,03 medical and health sciences ,humoral immunity ,medicine ,Immunology and Allergy ,Epstein-Barr virus ,Memory B cell ,B cell ,herpes virus ,Cell Biology ,medicine.disease ,Epstein–Barr virus ,Virology ,Vaccination ,030104 developmental biology ,medicine.anatomical_structure ,Humoral immunity ,Generation R ,pediatric infection - Abstract
The EBV is known to persist in memory B cells, but it remains unclear how this affects cell numbers and humoral immunity. We here studied EBV persistence in memory B cell subsets and consequences on B cell memory in young children. EBV genome loads were quantified in 6 memory B cell subsets in EBV+ adults. The effects of EBV infection on memory B cell numbers and vaccination responses were studied longitudinally in children within the Generation R population cohort between 14 mo and 6 yr of age. EBV genomes were more numerous in CD27+IgG+, CD27+IgA+, and CD27−IgA+ memory B cells than in IgM-only, natural effector, and CD27−IgG+ B cells. The blood counts of IgM-only, CD27+IgA+, CD27−IgG+, and CD27+IgG+ memory B cells were significantly lower in EBV+ children than in uninfected controls at 14 mo of age—the age when these cells peak in numbers. At 6 yr, all of these memory B cell counts had normalized, as had plasma IgG levels to previous primary measles and booster tetanus vaccinations. In conclusion, EBV persists predominantly in Ig class-switched memory B cells, even when derived from T cell-independent responses (CD27−IgA+), and EBV infection results in a transient depletion of these cells in young children.
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- 2017
15. EBV BCL-2 homologue BHRF1 drives chemoresistance and lymphomagenesis by inhibiting multiple cellular pro-apoptotic proteins
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Gemma L. Kelly, Marc Kvansakul, Clare Shannon-Lowe, Rosemary J. Tierney, Martin Rowe, Alan B. Rickinson, Laura C. A. Galbraith, D. Croom-Carter, Marco J Herold, Catherine Chang, Grant Dewson, Peter M. Colman, Rachel Cartlidge, Nenad Sejic, Andrew I. Bell, Andreas Strasser, Leah Fitzsimmons, David C.S. Huang, and Chathura D. Suraweera
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0301 basic medicine ,medicine.medical_specialty ,Carcinogenesis ,Apoptosis ,medicine.disease_cause ,Article ,Proto-Oncogene Proteins c-myc ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,Viral Proteins ,0302 clinical medicine ,Loss of Function Mutation ,Puma ,Internal medicine ,hemic and lymphatic diseases ,Cell Line, Tumor ,Virus latency ,medicine ,Animals ,Humans ,Progenitor cell ,Molecular Biology ,Hematology ,biology ,Bcl-2-Like Protein 11 ,Cell Death ,Sequence Homology, Amino Acid ,Venetoclax ,Cell Biology ,biology.organism_classification ,medicine.disease ,Burkitt Lymphoma ,Virus Latency ,Mice, Inbred C57BL ,Haematopoiesis ,030104 developmental biology ,chemistry ,Proto-Oncogene Proteins c-bcl-2 ,Cytoprotection ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Cancer research ,Protein Binding - Abstract
Epstein–Barr virus (EBV), which is ubiquitous in the adult population, is causally associated with human malignancies. Like many infectious agents, EBV has evolved strategies to block host cell death, including through expression of viral homologues of cellular BCL-2 pro-survival proteins (vBCL-2s), such as BHRF1. Small molecule inhibitors of the cellular pro-survival BCL-2 family proteins, termed ‘BH3-mimetics’, have entered clinical trials for blood cancers with the BCL-2 inhibitor venetoclax already approved for treatment of therapy refractory chronic lymphocytic leukaemia and acute myeloid leukaemia in the elderly. The generation of BH3-mimetics that could specifically target vBCL-2 proteins may be an attractive therapeutic option for virus-associated cancers, since these drugs would be expected to only kill virally infected cells with only minimal side effects on normal healthy tissues. To achieve this, a better understanding of the contribution of vBCL-2 proteins to tumorigenesis and insights into their biochemical functions is needed. In the context of Burkitt lymphoma (BL), BHRF1 expression conferred strong resistance to diverse apoptotic stimuli. Furthermore, BHRF1 expression in mouse haematopoietic stem and progenitor cells accelerated MYC-induced lymphoma development in a model of BL. BHRF1 interacts with the cellular pro-apoptotic BCL-2 proteins, BIM, BID, PUMA and BAK, but its capability to inhibit apoptosis could not be mapped solely to one of these interactions, suggesting plasticity is a key feature of BHRF1. Site-directed mutagenesis revealed a site in BHRF1 that was critical for its interaction with PUMA and blocking DNA-damage-induced apoptosis, identifying a potentially therapeutically targetable vulnerability in BHRF1.
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- 2019
16. The Global Landscape of EBV-Associated Tumors
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Claire Shannon-Lowe and Alan B. Rickinson
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0301 basic medicine ,Cancer Research ,lymphoma ,Review ,Biology ,carcinoma ,medicine.disease_cause ,lcsh:RC254-282 ,Virus ,03 medical and health sciences ,0302 clinical medicine ,hemic and lymphatic diseases ,medicine ,Epstein-Barr virus ,lymphoproliferative diseases ,B cell ,latency ,Cancer ,medicine.disease ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Epstein–Barr virus ,3. Good health ,Lymphoma ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Primary effusion lymphoma ,Diffuse large B-cell lymphoma ,Plasmablastic lymphoma - Abstract
Epstein-Barr virus (EBV), a gamma-1 herpesvirus, is carried as a life-long asymptomatic infection by the great majority of individuals in all human populations. Yet this seemingly innocent virus is aetiologically linked to two pre-malignant lymphoproliferative diseases (LPDs) and up to nine distinct human tumors; collectively these have a huge global impact, being responsible for some 200,000 new cases of cancer arising worldwide each year. EBV replicates in oral epithelium but persists as a latent infection within the B cell system and several of its diseases are indeed of B cell origin; these include B-LPD of the immunocompromised, Hodgkin Lymphoma (HL), Burkitt Lymphoma (BL), Diffuse Large B cell Lymphoma (DLBCL) and two rarer tumors associated with profound immune impairment, plasmablastic lymphoma (PBL) and primary effusion lymphoma (PEL). Surprisingly, the virus is also linked to tumors arising in other cellular niches which, rather than being essential reservoirs of virus persistence in vivo, appear to represent rare cul-de-sacs of latent infection. These non-B cell tumors include LPDs and malignant lymphomas of T or NK cells, nasopharyngeal carcinoma (NPC) and gastric carcinoma of epithelial origin, and leiomyosarcoma, a rare smooth muscle cell tumor of the immunocompromised. Here we describe the main characteristics of these tumors, their distinct epidemiologies, histological features and degrees of EBV association, then consider how their different patterns of EBV latency may reflect the alternative latency programmes through which the virus first colonizes and then persists in immunocompetent host. For each tumor, we discuss current understanding of EBV's role in the oncogenic process, the identity (where known) of host genetic and environmental factors predisposing tumor development, and the recent evidence from cancer genomics identifying somatic changes that either complement or in some cases replace the contribution of the virus. Thereafter we look for possible connections between the pathogenesis of these apparently different malignancies and point to new research areas where insights may be gained.
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- 2019
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17. Proteome-wide analysis of CD8+ T cell responses to EBV lytic infection
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Alan B. Rickinson, Andrew D. Hislop, Calum Forrest, and Jianmin Zuo
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medicine.anatomical_structure ,Lytic cycle ,Immunity ,T cell ,medicine ,Cytotoxic T cell ,General Materials Science ,Human leukocyte antigen ,Biology ,Virology ,In vitro ,CD8 ,Virus - Abstract
Epstein-Barr virus (EBV) is one of 9 Human herpesviruses that can develop lifelong persistence. These viruses provide an antigenically complex challenge that induce strong CD8+T cell immunity during primary infection and continue to shape this immunity through recurrent lytic reactivation. Here is described the first lytic proteome-wide analysis of CD8+T cell responses to EBV and the first to compare primary vs memory CD8+T cell responses to any human herpesvirus. Primary CD8+T cells were mitogenically expanded directly from the blood of infectious mononucleosis (IM) patients. Comparatively, memory CD8+T cells required pre-enrichment using autologous dendritic cells loaded with a lytically-infected EBV cell lysate and FACS selection based upon the activation marker 4-1BB. Enriched cells were then expanded in vitro as for IM cells. Preparations from 7 IM patients and 7 healthy carriers were screened against each of the 70 EBV lytic cycle proteins in combination with the donors’ HLA-I alleles. Multiple reactivities were identified across the full lytic cycle with 146 responses identified amongst the 7 IM patients and 96 amongst the 7 healthy carriers. However the distribution of responses varied between the 2 cohorts with primary responses targeting IE and a small group of E proteins whereas memory responses targeted all phases but with some prominent responses against L proteins. This infers that responses in primary infection therefore appear to be shaped by presentation on the infected cell surface prior to the activity of viral evasins. However long-term carriage appears to re-shape the virus-specific response.
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- 2019
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18. Impaired Epstein-Barr Virus-Specific Neutralizing Antibody Response during Acute Infectious Mononucleosis Is Coincident with Global B-Cell Dysfunction
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Alan B. Rickinson, Nick Tellam, Henry H. Balfour, Denis J. Moss, Michelle N. Wykes, Kristin A. Hogquist, Vijayendra Dasari, Rajiv Khanna, Corey Smith, Archana Panikkar, and Andrew D. Hislop
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Mononucleosis ,Tumor Necrosis Factor Ligand Superfamily Member 13 ,Immunology ,CD8-Positive T-Lymphocytes ,Antibodies, Viral ,Lymphocyte Activation ,medicine.disease_cause ,Microbiology ,Viral Matrix Proteins ,hemic and lymphatic diseases ,Virology ,B-Cell Activating Factor ,medicine ,Humans ,Infectious Mononucleosis ,fas Receptor ,B-cell activating factor ,Neutralizing antibody ,Epstein–Barr virus infection ,B cell ,B-Lymphocytes ,biology ,Immune dysregulation ,medicine.disease ,Antibodies, Neutralizing ,Epstein–Barr virus ,medicine.anatomical_structure ,Insect Science ,biology.protein ,Pathogenesis and Immunity ,Capsid Proteins ,Antibody ,Immunologic Memory - Abstract
Here we present evidence for previously unappreciated B-cell immune dysregulation during acute Epstein-Barr virus (EBV)-associated infectious mononucleosis (IM). Longitudinal analyses revealed that patients with acute IM have undetectable EBV-specific neutralizing antibodies and gp350-specific B-cell responses, which were associated with a significant reduction in memory B cells and no evidence of circulating antibody-secreting cells. These observations correlate with dysregulation of tumor necrosis factor family members BAFF and APRIL and increased expression of FAS on circulating B cells.
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- 2015
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19. The Immunology of Epstein-Barr Virus–Induced Disease
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Heather M. Long, Graham S. Taylor, Andrew D. Hislop, Jill Brooks, and Alan B. Rickinson
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Mononucleosis ,Immunology ,Disease ,Adaptive Immunity ,Biology ,medicine.disease_cause ,Virus ,Immunocompromised Host ,Immune system ,Immunopathology ,medicine ,Animals ,Humans ,Immunology and Allergy ,Immunodeficiency ,Immunologic Deficiency Syndromes ,medicine.disease ,Epstein–Barr virus ,Virology ,Immunity, Innate ,Lymphoproliferative Disorders ,Carrier State ,Hemophagocytosis - Abstract
Epstein-Barr virus (EBV) is usually acquired silently early in life and carried thereafter as an asymptomatic infection of the B lymphoid system. However, many circumstances disturb the delicate EBV-host balance and cause the virus to display its pathogenic potential. Thus, primary infection in adolescence can manifest as infectious mononucleosis (IM), as a fatal illness that magnifies the immunopathology of IM in boys with the X-linked lymphoproliferative disease trait, and as a chronic active disease leading to life-threatening hemophagocytosis in rare cases of T or natural killer (NK) cell infection. Patients with primary immunodeficiencies affecting the NK and/or T cell systems, as well as immunosuppressed transplant recipients, handle EBV infections poorly, and many are at increased risk of virus-driven B-lymphoproliferative disease. By contrast, a range of other EBV-positive malignancies of lymphoid or epithelial origin arise in individuals with seemingly intact immune systems through mechanisms that remain to be understood.
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- 2015
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20. The Epstein-Barr Virus BamHI C Promoter Is Not Essential for B Cell Immortalization In Vitro , but It Greatly Enhances B Cell Growth Transformation
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Rosemary J. Tierney, Jasdeep Nagra, Andrew I. Bell, Alan B. Rickinson, and Martin Rowe
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Herpesvirus 4, Human ,Transcription, Genetic ,viruses ,Immunology ,Response element ,Biology ,medicine.disease_cause ,Recombinant virus ,Microbiology ,Virus ,Viral Proteins ,hemic and lymphatic diseases ,Virology ,medicine ,Humans ,Promoter Regions, Genetic ,Transcription factor ,B cell ,Cell Proliferation ,B-Lymphocytes ,Cell growth ,Promoter ,Cell Transformation, Viral ,Epstein–Barr virus ,Molecular biology ,Virus-Cell Interactions ,medicine.anatomical_structure ,Insect Science ,Host-Pathogen Interactions ,Protein Binding - Abstract
Epstein-Barr virus (EBV) infection of B cells leads to the sequential activation of two viral promoters, Wp and Cp, resulting in the expression of six EBV nuclear antigens (EBNAs) and the viral Bcl2 homologue BHRF1. The viral transactivator EBNA2 is required for this switch from Wp to Cp usage during the initial stages of infection. EBNA2-dependent Cp transcription is mediated by the EBNA2 response element (E2RE), a region that contains at least two binding sites for cellular factors; one of these sites, CBF1, interacts with RBP-JK, which then recruits EBNA2 to the transcription initiation complex. Here we demonstrate that the B cell-specific transcription factor BSAP/Pax5 binds to a second site, CBF2, in the E2RE. Deletion of the E2RE in the context of a recombinant virus greatly diminished levels of Cp-initiated transcripts during the initial stages of infection but did not affect the levels of Wp-initiated transcripts or EBNA mRNAs. Consistent with this finding, viruses deleted for the E2RE were not markedly impaired in their ability to induce B cell transformation in vitro . In contrast, a larger deletion of the entire Cp region did reduce EBNA mRNA levels early after infection and subsequently almost completely ablated lymphoblastoid cell line (LCL) outgrowth. Notably, however, rare LCLs could be established following infection with Cp-deleted viruses, and these were indistinguishable from wild-type-derived LCLs in terms of steady-state EBV gene transcription. These data indicate that, unlike Wp, Cp is dispensable for the virus' growth-transforming activity. IMPORTANCE Epstein-Barr virus (EBV), a B lymphotropic herpesvirus etiologically linked to several B cell malignancies, efficiently induces B cell proliferation leading to the outgrowth of lymphoblastoid cell lines (LCLs). The initial stages of this growth-transforming infection are characterized by the sequential activation of two viral promoters, Wp and Cp, both of which appear to be preferentially active in target B cells. In this work, we have investigated the importance of Cp activity in initiating B cell proliferation and maintaining LCL growth. Using recombinant viruses, we demonstrate that while Cp is not essential for LCL outgrowth in vitro , it enhances transformation efficiency by >100-fold. We also show that Cp, like Wp, interacts with the B cell-specific activator protein BSAP/Pax5. We suggest that EBV has evolved this two-promoter system to ensure efficient colonization of the host B cell system in vivo .
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- 2015
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21. Epstein-Barr virus-associated lymphomas
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Claire, Shannon-Lowe, Alan B, Rickinson, and Andrew I, Bell
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Lymphoma ,Carcinogenesis ,Burkitt lymphoma ,Articles ,Review Article ,T/NK lymphoma ,diffuse large B cell lymphoma ,Epstein–Barr virus ,Immunocompromised Host ,hemic and lymphatic diseases ,post-transplant lymphoproliferative disease ,Humans ,Immunocompetence ,Hodgkin lymphoma - Abstract
Epstein–Barr virus (EBV), originally discovered through its association with Burkitt lymphoma, is now aetiologically linked to a remarkably wide range of lymphoproliferative lesions and malignant lymphomas of B-, T- and NK-cell origin. Some occur as rare accidents of virus persistence in the B lymphoid system, while others arise as a result of viral entry into unnatural target cells. The early finding that EBV is a potent B-cell growth transforming agent hinted at a simple oncogenic mechanism by which this virus could promote lymphomagenesis. In reality, the pathogenesis of EBV-associated lymphomas involves a complex interplay between different patterns of viral gene expression and cellular genetic changes. Here we review recent developments in our understanding of EBV-associated lymphomagenesis in both the immunocompetent and immunocompromised host. This article is part of the themed issue ‘Human oncogenic viruses’.
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- 2017
22. Asymptomatic Primary Infection with Epstein-Barr Virus: Observations on Young Adult Cases
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Rachel J, Abbott, Annette, Pachnio, Isabela, Pedroza-Pacheco, Alison M, Leese, Jusnara, Begum, Heather M, Long, Debbie, Croom-Carter, Andrea, Stacey, Paul A H, Moss, Andrew D, Hislop, Persephone, Borrow, Alan B, Rickinson, and Andrew I, Bell
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Adult ,Male ,B-Lymphocytes ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,infectious mononucleosis ,CD8-Positive T-Lymphocytes ,Viral Load ,Antibodies, Viral ,Prognosis ,host immune response ,United Kingdom ,Killer Cells, Natural ,Young Adult ,CD8 T cell ,DNA, Viral ,primary infection ,Humans ,Pathogenesis and Immunity ,Epstein-Barr virus ,Female ,NK cell ,Prospective Studies ,Asymptomatic Infections - Abstract
Epstein-Barr virus (EBV) is typically acquired asymptomatically in childhood. In contrast, infection later in life often leads to infectious mononucleosis (IM), a febrile illness characterized by anti-EBV IgM antibody positivity, high loads of circulating latently infected B cells, and a marked lymphocytosis caused by hyperexpansion of EBV-specific CD8+ T cells plus a milder expansion of CD56dim NKG2A+ KIR− natural killer (NK) cells. How the two situations compare is unclear due to the paucity of studies on clinically silent infection. Here we describe five prospectively studied patients with asymptomatic infections identified in a seroepidemiologic survey of university entrants. In each case, the key blood sample had high cell-associated viral loads without a marked CD8 lymphocytosis or NK cell disturbance like those seen in patients during the acute phase of IM. Two of the cases with the highest viral loads showed a coincident expansion of activated EBV-specific CD8+ T cells, but overall CD8+ T cell numbers were either unaffected or only mildly increased. Two cases with slightly lower loads, in whom serology suggests the infection may have been caught earlier in the course of infection, also showed no T or NK cell expansion at the time. Interestingly, in another case with a higher viral load, in which T and NK cell responses were undetectable in the primary blood sample in which infection was detected, EBV-specific T cell responses did not appear until several months later, by which time the viral loads in the blood had already fallen. Thus, some patients with asymptomatic primary infections have very high circulating viral loads similar to those in patients during the acute phase of IM and a cell-mediated immune response that is qualitatively similar to that in IM patients but of a lower magnitude. However, other patients may have quite different immune responses that ultimately could reveal novel mechanisms of host control. IMPORTANCE Epstein-Barr virus (EBV) is transmitted orally, replicates in the throat, and then invades the B lymphocyte pool through a growth-transforming latent infection. While primary infection in childhood is usually asymptomatic, delayed infection is associated with infectious mononucleosis (IM), a febrile illness in which patients have high circulating viral loads and an exaggerated virus-induced immune response involving both CD8+ T cells and natural killer (NK) cells. Here we show that in five cases of asymptomatic infection, viral loads in the blood were as high as those in patients during the acute phase of IM, whereas the cell-mediated responses, even when they resembled those in patients during the acute phase of IM in timing and quality, were never as exaggerated. We infer that IM symptoms arise as a consequence not of the virus infection per se but of the hyperactivated immune response. Interestingly, there were idiosyncratic differences among asymptomatic cases in the relationship between the viral load and the response kinetics, emphasizing how much there is still to learn about primary EBV infection.
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- 2017
23. A Recombinant Modified Vaccinia Ankara Vaccine Encoding Epstein–Barr Virus (EBV) Target Antigens: A Phase I Trial in UK Patients with EBV-Positive Cancer
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Hui Jia, Anthony T.C. Chan, David Price, Kevin J. Harrington, Lip Wai Lee, Graham S. Taylor, James E. Turner, Alan B. Rickinson, Lesley McGuigan, Edwin P. Hui, Jen Matthews, Neil Steven, Kristin Ladell, Andrew Hartley, Manjit Tanday, Ceri Edwards, Steve Wilson, and Claudia Roberts
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Adult ,Male ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Cancer Research ,Modified vaccinia Ankara ,viruses ,Epitopes, T-Lymphocyte ,T-Cell Antigen Receptor Specificity ,Vaccinia virus ,Biology ,medicine.disease_cause ,Cancer Vaccines ,Article ,Virus ,Immunophenotyping ,RC0254 ,SDG 3 - Good Health and Well-being ,Antigen ,T-Lymphocyte Subsets ,Neoplasms ,otorhinolaryngologic diseases ,medicine ,Humans ,Lymphocyte Count ,Aged ,Neoplasm Staging ,Immunogenicity ,ELISPOT ,Vaccination ,Middle Aged ,Viral Load ,medicine.disease ,Combined Modality Therapy ,Epstein–Barr virus ,Virology ,stomatognathic diseases ,Treatment Outcome ,Epstein-Barr Virus Nuclear Antigens ,Oncology ,Nasopharyngeal carcinoma ,Immunology ,Female - Abstract
Purpose: Epstein–Barr virus (EBV) is associated with several cancers in which the tumor cells express EBV antigens EBNA1 and LMP2. A therapeutic vaccine comprising a recombinant vaccinia virus, MVA-EL, was designed to boost immunity to these tumor antigens. A phase I trial was conducted to demonstrate the safety and immunogenicity of MVA-EL across a range of doses. Experimental Design: Sixteen patients in the United Kingdom (UK) with EBV-positive nasopharyngeal carcinoma (NPC) received three intradermal vaccinations of MVA-EL at 3-weekly intervals at dose levels between 5 × 107 and 5 × 108 plaque-forming units (pfu). Blood samples were taken at screening, after each vaccine cycle, and during the post-vaccination period. T-cell responses were measured using IFNγ ELISpot assays with overlapping EBNA1/LMP2 peptide mixes or HLA-matched epitope peptides. Polychromatic flow cytometry was used to characterize functionally responsive T-cell populations. Results: Vaccination was generally well tolerated. Immunity increased after vaccination to at least one antigen in 8 of 14 patients (7/14, EBNA1; 6/14, LMP2), including recognition of epitopes that vary between EBV strains associated with different ethnic groups. Immunophenotypic analysis revealed that vaccination induced differentiation and functional diversification of responsive T-cell populations specific for EBNA1 and LMP2 within the CD4 and CD8 compartments, respectively. Conclusions: MVA-EL is safe and immunogenic across diverse ethnicities and thus suitable for use in trials against different EBV-positive cancers globally as well as in South-East Asia where NPC is most common. The highest dose (5 × 108 pfu) is recommended for investigation in current phase IB and II trials. Clin Cancer Res; 20(19); 5009–22. ©2014 AACR.
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- 2014
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24. Cellular immune controls over Epstein–Barr virus infection: new lessons from the clinic and the laboratory
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Alan B. Rickinson, Heather M. Long, Andrew D. Hislop, Christian Münz, Umaimainthan Palendira, University of Zurich, and Rickinson, Alan B
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Epstein-Barr Virus Infections ,Mononucleosis ,Immunology ,610 Medicine & health ,Biology ,10263 Institute of Experimental Immunology ,medicine.disease_cause ,Virus ,Immune system ,Immunopathology ,medicine ,Animals ,Humans ,Immunology and Allergy ,Epstein–Barr virus infection ,B cell ,Immunodeficiency ,2403 Immunology ,Immunity, Cellular ,medicine.disease ,Epstein–Barr virus ,Virology ,Disease Models, Animal ,medicine.anatomical_structure ,2723 Immunology and Allergy ,570 Life sciences ,biology - Abstract
Epstein-Barr virus (EBV), a human herpesvirus with potent B cell growth transforming ability, induces multiple cellular immune responses in the infected host. How these host responses work together to prevent virus pathogenicity, and how immune imbalance predisposes to disease, remain poorly understood. Here, we describe three ongoing lines of enquiry that are shedding new light on these issues. These focus on: (i) patients with infectious mononucleosis or its fatal equivalent, X-linked lymphoproliferative disease; (ii) EBV infection in a range of new, genetically defined, primary immune deficiency states; and (iii) experimental infection in two complementary animal models, the rhesus macaque and the human haemopoietic stem cell reconstituted mouse.
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- 2014
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25. MHC II tetramers visualize human CD4+ T cell responses to Epstein–Barr virus infection and demonstrate atypical kinetics of the nuclear antigen EBNA1 response
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Alan B. Rickinson, Odette Chagoury, Rachel J. M. Abbott, Shereen Sabbah, Alison M. Leese, Heather M. Long, Eddie A. James, William W. Kwok, Gordon B. Ryan, and Laura T. Morton
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CD4-Positive T-Lymphocytes ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,viruses ,T cell ,Immunology ,Priming (immunology) ,Streptamer ,Biology ,Article ,03 medical and health sciences ,Epitopes ,0302 clinical medicine ,Antigen ,Species Specificity ,hemic and lymphatic diseases ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Humans ,Infectious Mononucleosis ,Antigen-presenting cell ,Antigens, Viral ,030304 developmental biology ,Cell Proliferation ,0303 health sciences ,Histocompatibility Antigens Class II ,Convalescence ,MHC restriction ,Virology ,Molecular biology ,3. Good health ,Kinetics ,medicine.anatomical_structure ,Phenotype ,Epstein-Barr Virus Nuclear Antigens ,Immunoglobulin G ,Acute Disease ,Antibody Formation ,Protein Multimerization ,Immunologic Memory ,CD8 ,030215 immunology - Abstract
Characterization of the human EBV-specific CD4+ T cell response using MHC II tetramers reveals the latent EBV antigen response is more frequent than the lytic response with a delayed EBNA1 response that coincides with diminished cross-presentation., Virus-specific CD4+ T cells are key orchestrators of host responses to viral infection yet, compared with their CD8+ T cell counterparts, remain poorly characterized at the single cell level. Here we use nine MHC II–epitope peptide tetramers to visualize human CD4+ T cell responses to Epstein–Barr virus (EBV), the causative agent of infectious mononucleosis (IM), a disease associated with large virus-specific CD8+ T cell responses. We find that, while not approaching virus-specific CD8+ T cell expansions in magnitude, activated CD4+ T cells specific for epitopes in the latent antigen EBNA2 and four lytic cycle antigens are detected at high frequencies in acute IM blood. They then fall rapidly to values typical of life-long virus carriage where most tetramer-positive cells display conventional memory markers but some, unexpectedly, revert to a naive-like phenotype. In contrast CD4+ T cell responses to EBNA1 epitopes are greatly delayed in IM patients, in line with the well-known but hitherto unexplained delay in EBNA1 IgG antibody responses. We present evidence from an in vitro system that may explain these unusual kinetics. Unlike other EBNAs and lytic cycle proteins, EBNA1 is not naturally released from EBV-infected cells as a source of antigen for CD4+ T cell priming.
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- 2013
26. Compartmentalization of Total and Virus-Specific Tissue-Resident Memory CD8+ T Cells in Human Lymphoid Organs
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Andrew I. Bell, Jane Li, Andrew D. Hislop, Stuart G. Tangye, Heng Giap Woon, Umaimainthan Palendira, Alan B. Rickinson, Frederic Sierro, Warwick J. Britton, Asolina Braun, Corey Smith, Jarem Edwards, Carl G. Feng, Rajiv Khanna, Thomas Gebhardt, and Michael Elliot
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0301 basic medicine ,Male ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Physiology ,Cytomegalovirus ,CD8-Positive T-Lymphocytes ,Memory T cells ,Interleukin 21 ,White Blood Cells ,0302 clinical medicine ,Spectrum Analysis Techniques ,Animal Cells ,Immune Physiology ,Medicine and Health Sciences ,Cytotoxic T cell ,IL-2 receptor ,Biology (General) ,Interleukin-15 ,Innate Immune System ,T Cells ,ZAP70 ,Natural killer T cell ,Tonsils ,Flow Cytometry ,3. Good health ,Cell biology ,medicine.anatomical_structure ,Spectrophotometry ,Organ Specificity ,Cytomegalovirus Infections ,Cytokines ,Female ,Cytophotometry ,Cellular Types ,Anatomy ,Research Article ,QH301-705.5 ,T cell ,Immune Cells ,Immunology ,Kruppel-Like Transcription Factors ,Cytotoxic T cells ,Biology ,Research and Analysis Methods ,Microbiology ,Throat ,03 medical and health sciences ,Antigens, CD ,Virology ,Genetics ,medicine ,Humans ,Antigen-presenting cell ,Molecular Biology ,Interleukin 3 ,Blood Cells ,Biology and Life Sciences ,Cell Biology ,Molecular Development ,RC581-607 ,Viral Replication ,030104 developmental biology ,Immune System ,Parasitology ,Immunologic diseases. Allergy ,Immunologic Memory ,Neck ,Spleen ,030215 immunology ,Developmental Biology - Abstract
Disruption of T cell memory during severe immune suppression results in reactivation of chronic viral infections, such as Epstein Barr virus (EBV) and Cytomegalovirus (CMV). How different subsets of memory T cells contribute to the protective immunity against these viruses remains poorly defined. In this study we examined the compartmentalization of virus-specific, tissue resident memory CD8+ T cells in human lymphoid organs. This revealed two distinct populations of memory CD8+ T cells, that were CD69+CD103+ and CD69+CD103—, and were retained within the spleen and tonsils in the absence of recent T cell stimulation. These two types of memory cells were distinct not only in their phenotype and transcriptional profile, but also in their anatomical localization within tonsils and spleen. The EBV-specific, but not CMV-specific, CD8+ memory T cells preferentially accumulated in the tonsils and acquired a phenotype that ensured their retention at the epithelial sites where EBV replicates. In vitro studies revealed that the cytokine IL-15 can potentiate the retention of circulating effector memory CD8+ T cells by down-regulating the expression of sphingosine-1-phosphate receptor, required for T cell exit from tissues, and its transcriptional activator, Kruppel-like factor 2 (KLF2). Within the tonsils the expression of IL-15 was detected in regions where CD8+ T cells localized, further supporting a role for this cytokine in T cell retention. Together this study provides evidence for the compartmentalization of distinct types of resident memory T cells that could contribute to the long-term protection against persisting viral infections., Author Summary Some viruses have the capacity to establish chronic infections in humans. How different T cell populations effectively control these infections has not been clear. Continuous circulation of memory T cells was thought to be crucial for effective immune surveillance against such infections. Recent studies in mice however, have shown that non-circulating tissue resident memory populations can also contribute to protective immunity. In this study we have examined the distribution, localization and characteristics of Epstein-Barr virus and Cytomegalovirus-specific T cells in different human tissues. This showed that virus-specific T cells were differentially distributed in different tissues and there was preferential accumulation of EBV-specific resident memory T cells at sites where EBV reactivates. In vitro studies showed that IL-15 and TGF-β could cooperate to extinguish tissue exit signals in T cells and therefore potentiate their retention within tissues. IL-15 expression was also detected in areas where T cells aggregated within the tissue. Together our study provides insight into how distinct memory T cells are compartmentalized in tissues to maintain long-term protection against persisting viral infections.
- Published
- 2016
27. Reconstitution of the Epstein-Barr virus-specific cytotoxic T-lymphocyte response following T-cell-depleted myeloablative and nonmyeloablative allogeneic stem cell transplantation
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Donald Milligan, Alan B. Rickinson, Dorothy McDonald, Suparno Chakrabarti, Kathleen Holder, Stephen Mackinnon, Deenan Pillay, Narinder Kaur, Herman Waldmann, Christopher Fegan, Geoff Hale, and Neil Steven
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Melphalan ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Transplantation Conditioning ,Antibodies, Neoplasm ,medicine.medical_treatment ,T-Lymphocytes ,Immunology ,Graft vs Host Disease ,Hematopoietic stem cell transplantation ,Antibodies, Monoclonal, Humanized ,Biochemistry ,Lymphocyte Depletion ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Cytotoxic T cell ,Humans ,Transplantation, Homologous ,Alemtuzumab ,Antigens, Viral ,business.industry ,ELISPOT ,Hematopoietic Stem Cell Transplantation ,Antibodies, Monoclonal ,Cell Biology ,Hematology ,Fludarabine ,Transplantation ,Hematologic Neoplasms ,business ,medicine.drug ,T-Lymphocytes, Cytotoxic - Abstract
The recovery of circulating antigen-specific T-cell immunity to Epstein-Barr virus (EBV) was determined in ELIspot assays following allogeneic myeloablative or nonmyeloablative stem cell transplantation (MST/NST). In 8 of 12 MST patients receiving an alemtuzumab-treated graft, the frequency of the EBV-specific reactivities was similar to or greater than that seen in the healthy controls. A response was detectable in 3 of 6 and 6 of 9 patients by 3 and 6 months, respectively, and in all patients by one year following MST. In contrast, only 1 of 9 (95% confidence interval [CI], 0-2.8) patients made a detectable EBV-specific response by 6 months following NST conditioned with fludarabine, melphalan, and alemtuzumab. Responses were detected in 7 of 10 patients by 1 year after NST. Parallel surveillance demonstrated that other virus infections occurred more frequently and earlier after transplantation in NST patients. The use of alemtuzumab in vivo in the nonmyeloablative conditioning might have resulted in the delay in EBV-specific T-cell recovery and increased virus infections.
- Published
- 2016
28. Virus-specific cytotoxic T lymphocytes differentially express cell-surface leukocyte immunoglobulin-like receptor-1, an inhibitory receptor for class I major histocompatibility complex molecules
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Richard D Antrobus, Alan B. Rickinson, Paul Moss, Naeem Khan, Damien J. Montamat-Sicotte, Benjamin E. Willcox, Lee I. Garner, and Andrew D. Hislop
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CD4-Positive T-Lymphocytes ,Epstein-Barr Virus Infections ,Virus-specific Cytotoxic T-lymphocytes ,T cell ,viruses ,Leukocyte Immunoglobulin-like Receptor B1 ,Major histocompatibility complex ,Species Specificity ,Antigens, CD ,HLA Antigens ,Recurrence ,Influenza, Human ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Humans ,Receptors, Immunologic ,biology ,Effector ,CD28 ,Cell biology ,Infectious Diseases ,medicine.anatomical_structure ,Gene Expression Regulation ,Immunology ,Cytomegalovirus Infections ,biology.protein ,CD8 ,T-Lymphocytes, Cytotoxic - Abstract
Leukocyte immunoglobulin-like receptor-1 (LIR-1) is an inhibitory receptor that negatively regulates T cell effector functions after interaction with host class I major histocompatibility complex molecules and, additionally, binds to UL18, a human cytomegalovirus (HCMV)-encoded class I homologue. Here, we demonstrate that virus-specific cytotoxic T lymphocytes (CTLs) differentially express LIR-1, with high frequencies of expression on HCMV-specific CD8+ T cells and intermediate and low frequencies of expression on influenza virus-specific and Epstein-Barr virus (EBV)-specific CTLs, respectively. Expression of LIR-1 was dependent on CTL-antigen specificity and was associated with a differentiated effector memory phenotype, as demonstrated by decreased expression of CD28 and increased expression of CD57. During primary HCMV and EBV infections, expression of LIR-1 on virus-specific CTLs was low and increased slowly. These results indicate that expression of LIR-1 increases during differentiation of virus-specific CD8+ effector T cells. Furthermore, they suggest that a potential immunoregulatory function of UL18 may be to preferentially target highly differentiated HCMV-specific effector memory T cells during persistent infection.
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- 2016
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29. Transient reduction in IgA
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Diana, van den Heuvel, Michelle A E, Jansen, Andrew I, Bell, Alan B, Rickinson, Vincent W V, Jaddoe, Jacques J M, van Dongen, Henriette A, Moll, and Menno C, van Zelm
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Adult ,Male ,B-Lymphocytes ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Adolescent ,Vaccination ,Infant ,Lymphocyte Depletion ,Immunoglobulin A ,Immunoglobulin G ,Humans ,Female ,Lymphocyte Count ,Child ,Immunologic Memory - Abstract
The EBV is known to persist in memory B cells, but it remains unclear how this affects cell numbers and humoral immunity. We here studied EBV persistence in memory B cell subsets and consequences on B cell memory in young children. EBV genome loads were quantified in 6 memory B cell subsets in EBV
- Published
- 2016
30. Early T Cell Recognition of B Cells following Epstein-Barr Virus Infection: Identifying Potential Targets for Prophylactic Vaccination
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Martin Fitzpatrick, Alan B. Rickinson, Heather M. Long, Graham S. Taylor, Jill Brooks, Rose J. Tierney, Claire Shannon-Lowe, and Alison M. Leese
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0301 basic medicine ,Enzyme-Linked Immunospot Assay ,Epstein-Barr Virus Infections ,B Cells ,Epitopes, T-Lymphocyte ,Antigen Processing and Recognition ,CD8-Positive T-Lymphocytes ,Lymphocyte Activation ,Epitope ,White Blood Cells ,0302 clinical medicine ,Animal Cells ,hemic and lymphatic diseases ,Medicine and Health Sciences ,Cytotoxic T cell ,Biology (General) ,Enzyme-Linked Immunoassays ,Immune Response ,Antigens, Viral ,B-Lymphocytes ,T Cells ,3. Good health ,medicine.anatomical_structure ,Cellular Types ,Research Article ,Lytic Cycle ,QH301-705.5 ,T cell ,Immune Cells ,Naive B cell ,Immunology ,Immunoblotting ,Cytotoxic T cells ,Streptamer ,Biology ,Research and Analysis Methods ,Microbiology ,03 medical and health sciences ,Antigen ,Virology ,Genetics ,medicine ,Humans ,Antigen-presenting cell ,Antibody-Producing Cells ,Molecular Biology Techniques ,Immunoassays ,Molecular Biology ,Blood Cells ,Biology and Life Sciences ,Viral Vaccines ,Cell Biology ,RC581-607 ,Viral Replication ,B-1 cell ,030104 developmental biology ,Immunologic Techniques ,Parasitology ,Immunologic diseases. Allergy ,030215 immunology ,Cloning - Abstract
Epstein-Barr virus, a B-lymphotropic herpesvirus, is the cause of infectious mononucleosis, has strong aetiologic links with several malignancies and has been implicated in certain autoimmune diseases. Efforts to develop a prophylactic vaccine to prevent or reduce EBV-associated disease have, to date, focused on the induction of neutralising antibody responses. However, such vaccines might be further improved by inducing T cell responses capable of recognising and killing recently-infected B cells. In that context, EBNA2, EBNA-LP and BHRF1 are the first viral antigens expressed during the initial stage of B cell growth transformation, yet have been poorly characterised as CD8+ T cell targets. Here we describe CD8+ T cell responses against each of these three “first wave” proteins, identifying target epitopes and HLA restricting alleles. While EBNA-LP and BHRF1 each contained one strong CD8 epitope, epitopes within EBNA2 induced immunodominant responses through several less common HLA class I alleles (e.g. B*3801 and B*5501), as well as subdominant responses through common class I alleles (e.g. B7 and C*0304). Importantly, such EBNA2-specific CD8+ T cells recognised B cells within the first day post-infection, prior to CD8+ T cells against well-characterised latent target antigens such as EBNA3B or LMP2, and effectively inhibited outgrowth of EBV-transformed B cell lines. We infer that “first wave” antigens of the growth-transforming infection, especially EBNA2, constitute potential CD8+ T cell immunogens for inclusion in prophylactic EBV vaccine design., Author Summary Epstein-Barr virus infects the vast majority of the world’s population; in most individuals both primary infection and long-term virus carriage are asymptomatic. However, EBV is the major cause of glandular fever, is associated with multiple cancers and is implicated in various autoimmune conditions; thus there is a strong impetus for the development of a prophylactic vaccine. To date, vaccine design has largely focused on the induction of neutralising antibodies to virion structural components which can prevent virus binding and infection. Such strategies may be improved by the inclusion of immunogens to induce T cell responses with the potential to promptly recognise and eliminate cells that do become infected. Here we characterise T cell responses to three proteins, EBNA2, EBNA-LP and BHRF1 that comprise the “first wave” of de novo viral antigen expression following EBV infection of B cells. Each of these proteins is targeted by strong T cell responses in a subset of donors. Furthermore, CD8+ T cells specific for that at least one of these proteins, EBNA2, efficiently recognise B cells at very early time-points post-infection, before CD8+ T cells of all other specificities tested, and effectively inhibit outgrowth of B cell lines following EBV infection in vitro. Thus “first wave antigens”, particularly EBNA2, may comprise suitable candidate immunogens for inclusion in prophylactic EBV vaccine design.
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- 2016
31. Immunity to Oncogenic Viruses
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Nicholas Easom, Mala K. Maini, Alan B. Rickinson, and Martin Rowe
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medicine.medical_treatment ,fungi ,food and beverages ,Immunotherapy ,Biology ,Acquired immune system ,medicine.disease_cause ,Epstein–Barr virus ,Virology ,Vaccination ,Immune system ,Immunity ,Immunology ,medicine ,Carcinogenesis ,Oncovirus - Abstract
In this article, we review the oncogenic viruses that together cause 10% of all cancer cases worldwide. The responsible viruses are from diverse families but can all escape host immunity to establish persistent, often lifelong, infection. The resultant infections can remain asymptomatic or can have a number of consequences, only driving oncogenic changes in a minority of individuals. In many cases, individual viral components have roles in promoting tumor formation by integration into the host genome and in evasion of innate and adaptive immunity. However, the immune responses these viruses do elicit can also contribute to oncogenesis through chronic inflammation and/or can restrain tumor formation through active immune surveillance, as evidenced by the increased emergence of several of these tumors in the setting of immunosuppression. Here we summarize current knowledge of the immune responses to oncogenic viruses and their associated tumors, highlighting prospects for harnessing immunity in prevention and treatment.
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- 2016
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32. Expansion of somatically reverted memory CD8+ T cells in patients with X-linked lymphoproliferative disease caused by selective pressure from Epstein-Barr virus
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Sharon Choo, Rebecca H. Buckley, Alan B. Rickinson, Stephen Adelstein, Bodo Grimbacher, Rachel J. M. Abbott, Umaimainthan Palendira, Joanne Smart, Stuart G. Tangye, Carol Low, Cindy S. Ma, Amy D. Klion, Kim E. Nichols, Silvia Giliani, Andrew I. Bell, Frank Alvaro, Tri Giang Phan, Vassilios Lougaris, and D. Sean Riminton
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Herpesvirus 4, Human ,Immunology ,Molecular Sequence Data ,Lymphoproliferative disorders ,Biology ,CD8-Positive T-Lymphocytes ,medicine.disease_cause ,Virus ,hemic and lymphatic diseases ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Humans ,Amino Acid Sequence ,Signaling Lymphocytic Activation Molecule Associated Protein ,Epstein–Barr virus infection ,DNA Primers ,Base Sequence ,Brief Definitive Report ,Intracellular Signaling Peptides and Proteins ,X-linked lymphoproliferative disease ,Exons ,Sequence Analysis, DNA ,Viral Load ,medicine.disease ,Flow Cytometry ,Epstein–Barr virus ,Virology ,Lymphoproliferative Disorders ,Gene Expression Regulation ,Mutation ,Primary immunodeficiency ,Immunologic Memory ,CD8 - Abstract
In patients with XLP, a primary immunodeficiency caused by mutations in SH2D1A, EBV infection can lead to somatic reversion of the disease-causing mutation selectively in effector memory CD8 T cells; reverted CD8 cells are better able to respond to and kill EBV-infected cells., Patients with the primary immunodeficiency X-linked lymphoproliferative disease (XLP), which is caused by mutations in SH2D1A, are highly susceptible to Epstein-Barr virus (EBV) infection. Nonetheless, some XLP patients demonstrate less severe clinical manifestations after primary infection. SH2D1A encodes the adaptor molecule SLAM-associated protein (SAP), which is expressed in T and natural killer cells and is required for cytotoxicity against B cells, the reservoir for EBV. It is not known why the clinical presentation of XLP is so variable. In this study, we report for the first time the occurrence of somatic reversion in XLP. Reverted SAP-expressing cells resided exclusively within the CD8+ T cell subset, displayed a CD45RA−CCR7− effector memory phenotype, and were maintained at a stable level over time. Importantly, revertant CD8+ SAP+ T cells, but not SAP− cells, proliferated in response to EBV and killed EBV-infected B cells. As somatic reversion correlated with EBV infection, we propose that the virus exerts a selective pressure on the reverted cells, resulting in their expansion in vivo and host protection against ongoing infection.
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- 2012
33. Structural and energetic evidence for highly peptide-specific tumor antigen targeting via allo-MHC restriction
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Neil Steven, Alan B. Rickinson, Fiyaz Mohammed, Paul Moss, Amy Tranter, Hans J. Stauss, Amy A. Simpson, Mahboob Salim, and Benjamin E. Willcox
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Multidisciplinary ,biology ,Antigen processing ,T-Lymphocytes ,T-cell receptor ,Receptors, Antigen, T-Cell ,Biological Sciences ,MHC restriction ,Crystallography, X-Ray ,Major histocompatibility complex ,Hodgkin Disease ,Tumor antigen ,Cell biology ,Major Histocompatibility Complex ,Antigen ,Antigens, Neoplasm ,HLA-A2 Antigen ,Immunology ,biology.protein ,Humans ,Immunotherapy ,Crystallization ,Peptides ,Allorecognition - Abstract
Immunotherapies targeting peptides presented by allogeneic MHC molecules offer the prospect of circumventing tolerance to key tumor-associated self-antigens. However, the degree of antigen specificity mediated by alloreactive T cells, and their ability to discriminate normal tissues from transformed cells presenting elevated antigen levels, is poorly understood. We examined allorecognition of an HLA-A2–restricted Hodgkin's lymphoma-associated antigen and were able to isolate functionally antigen-specific allo-HLA-A2–restricted T cells from multiple donors. Binding and structural studies, focused on a prototypic allo-HLA-A2–restricted T-cell receptor (TCR) termed NB20 derived from an HLA-A3 homozygote, suggested highly peptide-specific allorecognition that was energetically focused on antigen, involving direct recognition of a distinct allopeptide presented within a conserved MHC recognition surface. Although NB20/HLA-A2 affinity was unremarkable, TCR/MHC complexes were very short-lived, consistent with suboptimal TCR triggering and tolerance to low antigen levels. These data provide strong molecular evidence that within the functionally heterogeneous alloreactive repertoire, there is the potential for highly antigen-specific “allo-MHC–restricted” recognition and suggest a kinetic mechanism whereby allo-MHC–restricted T cells may discriminate normal from transformed tissue, thereby outlining a suitable basis for broad-based therapeutic targeting of tolerizing tumor antigens.
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- 2011
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34. Epstein-Barr Virus BamHI W Repeat Number Limits EBNA2/EBNA-LP Coexpression in Newly Infected B Cells and the Efficiency of B-Cell Transformation: a Rationale for the Multiple W Repeats in Wild-Type Virus Strains
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Kuan-Yu Kao, Rosemary J. Tierney, Jasdeep Nagra, and Alan B. Rickinson
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,viruses ,Immunology ,Mutant ,Biology ,Real-Time Polymerase Chain Reaction ,medicine.disease_cause ,Microbiology ,Genome ,Virus ,law.invention ,Viral Proteins ,law ,hemic and lymphatic diseases ,Virology ,medicine ,Humans ,Gene ,Cells, Cultured ,Repetitive Sequences, Nucleic Acid ,Genetics ,B-Lymphocytes ,Cell Transformation, Viral ,Epstein–Barr virus ,Virus-Cell Interactions ,Transformation (genetics) ,Epstein-Barr Virus Nuclear Antigens ,Insect Science ,Recombinant DNA ,RNA, Viral ,BamHI - Abstract
The genome of Epstein-Barr virus (EBV), a gammaherpesvirus with potent B-cell growth-transforming ability, contains multiple copies of a 3-kb BamHI W repeat sequence; each repeat carries (i) a promoter (Wp) that initiates transformation by driving EBNA-LP and EBNA2 expression and (ii) the W1W2 exons encoding the functionally active repeat domain of EBNA-LP. The W repeat copy number of a virus therefore influences two potential determinants of its transforming ability: the number of available Wp copies and the maximum size of the encoded EBNA-LP. Here, using recombinant EBVs, we show that optimal B-cell transformation requires a minimum of 5 W repeats (5W); the levels of transforming ability fall progressively with viruses carrying 4, 3, and 2 W repeats, as do the levels of Wp-initiated transcripts expressed early postinfection (p.i.), while viruses with 1 copy of the wild-type W repeat (1W) and 0W are completely nontransforming. We therefore suggest that genetic analyses of EBV transforming function should ensure that wild-type and mutant strains have equal numbers (ideally at least 5) of W copies if the analysis is not to be compromised. Attempts to enhance the transforming function of low-W-copy-number viruses, via the activity of helper EBV strains or by gene repair, suggested that the critical defect is not related to EBNA-LP size but to the failure to achieve sufficiently strong coexpression of EBNA-LP and EBNA2 early postinfection. We further show by the results of ex vivo assays that EBV strains in the blood of infected individuals typically have a mean of 5 to 8 W copies, consistent with the view that evolution has selected for viruses with an optimal transforming function.
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- 2011
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35. The role of tetraspanin CD63 in antigen presentation via MHC class II
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Fedor Berditchevski, Alan B. Rickinson, Sven H. Petersen, Tracey A. Haigh, Graham S. Taylor, and Elena Odintsova
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CD4-Positive T-Lymphocytes ,Herpesvirus 4, Human ,CD74 ,Immunology ,CD1 ,Adaptive Immunity ,Exosomes ,Lymphocyte Activation ,hemic and lymphatic diseases ,MHC class I ,Humans ,Immunology and Allergy ,Cytotoxic T cell ,RNA, Small Interfering ,Antigens, Viral ,Cell Line, Transformed ,Antigen Presentation ,B-Lymphocytes ,MHC class II ,biology ,Tetraspanin 30 ,Antigen processing ,Histocompatibility Antigens Class II ,Transporter associated with antigen processing ,MHC restriction ,Molecular biology ,Clone Cells ,Microscopy, Electron ,biology.protein - Abstract
Interactions between MHC class II (MHC II)-positive APCs and CD4(+) T cells are central to adaptive immune responses. Using an Epstein-Barr virus (EBV)-transformed B lymphoblastoid cell line (LCL) as MHC II-positive APCs and CD4(+) T-cell clones specific for two endogenously expressed EBV antigens, we found that shRNA knockdown of the tetraspanin protein CD63 in LCL cells consistently led to increased CD4(+) T-cell recognition. This effect was not due to enhanced antigen processing nor to changes in MHC II expression since CD63 knockdown did not influence the amount or dimerization of MHC II in LCL cells. We therefore investigated the possible involvement of exosomes, small MHC II- and tetraspanin-abundant vesicles which are secreted by LCL cells and which we found could themselves activate the CD4(+) T-cell clones in an MHC II-dependent manner. While equal loadings of exosomes purified from the control and CD63(low) LCLs stimulated T cells to a comparable degree, we found that exosome production significantly increased following CD63-knockdown, suggesting that this may underlie the greater T-cell stimulatory capacity of the CD63(low) LCLs. Taken together, our data reveal a new insight into the mechanisms by which tetraspanins are involved in the regulation of MHC II-dependent T-cell stimulation.
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- 2011
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36. Epstein–Barr virus latent gene sequences as geographical markers of viral origin: unique EBNA3 gene signatures identify Japanese viruses as distinct members of the Asian virus family
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Maho Koyama-Sato, Masahiro Yasui, Alan B. Rickinson, D. Croom-Carter, Keisei Kawa, Akihisa Sawada, Rosemary J. Tierney, Osamu Kondo, and Masami Inoue
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Genotype ,Sequence analysis ,Population ,Biology ,medicine.disease_cause ,Virus ,Herpesviridae ,Viral Matrix Proteins ,hemic and lymphatic diseases ,Virology ,medicine ,Humans ,Allele ,education ,Gene ,Genetics ,education.field_of_study ,Polymorphism, Genetic ,Sequence Analysis, DNA ,Epstein–Barr virus ,Phylogeography ,Epstein-Barr Virus Nuclear Antigens ,DNA, Viral - Abstract
Polymorphisms in Epstein–Barr virus (EBV) latent genes can identify virus strains from different human populations and individual strains within a population. An Asian EBV signature has been defined almost exclusively from Chinese viruses, with little information from other Asian countries. Here we sequenced polymorphic regions of the EBNA1, 2, 3A, 3B, 3C and LMP1 genes of 31 Japanese strains from control donors and EBV-associated T/NK-cell lymphoproliferative disease (T/NK-LPD) patients. Though identical to Chinese strains in their dominant EBNA1 and LMP1 alleles, Japanese viruses were subtly different at other loci. Thus, while Chinese viruses mainly fall into two families with strongly linked ‘Wu’ or ‘Li’ alleles at EBNA2 and EBNA3A/B/C, Japanese viruses all have the consensus Wu EBNA2 allele but fall into two families at EBNA3A/B/C. One family has variant Li-like sequences at EBNA3A and 3B and the consensus Li sequence at EBNA3C; the other family has variant Wu-like sequences at EBNA3A, variants of a low frequency Chinese allele ‘Sp’ at EBNA3B and a consensus Sp sequence at EBNA3C. Thus, EBNA3A/B/C allelotypes clearly distinguish Japanese from Chinese strains. Interestingly, most Japanese viruses also lack those immune-escape mutations in the HLA-A11 epitope-encoding region of EBNA3B that are so characteristic of viruses from the highly A11-positive Chinese population. Control donor-derived and T/NK-LPD-derived strains were similarly distributed across allelotypes and, by using allelic polymorphisms to track virus strains in patients pre- and post-haematopoietic stem-cell transplant, we show that a single strain can induce both T/NK-LPD and B-cell-lymphoproliferative disease in the same patient.
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- 2011
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37. Immune defence against EBV and EBV-associated disease
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Heather M. Long, Alan B. Rickinson, and Graham S. Taylor
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Mononucleosis ,T-Lymphocytes ,T cell ,Immunology ,Population ,Disease ,Human leukocyte antigen ,Biology ,Asymptomatic ,Virus ,Immune system ,Histocompatibility Antigens ,Neoplasms ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,Immunology and Allergy ,education ,education.field_of_study ,Virus Internalization ,medicine.disease ,Virology ,medicine.anatomical_structure ,medicine.symptom - Abstract
Epstein-Barr virus (EBV), a B-lymphotropic herpesvirus widespread in the human population and normally contained as an asymptomatic infection by T cell surveillance, nevertheless causes infectious mononucleosis and is strongly linked to several types of human cancer. Here we describe new findings on the range of cellular immune responses induced by EBV infection, on viral strategies to evade those responses and on the links between HLA gene loci and EBV-induced disease. The success of adoptive T cell therapy for EBV-driven post-transplant lymphoproliferative disease is stimulating efforts to target other EBV-associated tumours by immunotherapeutic means, and has reawakened interest in the ultimate intervention strategy, a prophylactic EBV vaccine.
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- 2011
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38. A novel latent membrane 2 transcript expressed in Epstein-Barr virus–positive NK- and T-cell lymphoproliferative disease encodes a target for cellular immunotherapy
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Tracey A. Haigh, Wing C. Chan, Andrew I. Bell, Simon O'Connor, Alan B. Rickinson, Heather M. Long, Catherine M. Bollard, Martin Rowe, Javeed Iqbal, Christopher P. Fox, Claire Shannon-Lowe, Steven P. Lee, and Graham S. Taylor
- Subjects
Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Genes, Viral ,T-Lymphocytes ,T cell ,medicine.medical_treatment ,Immunology ,Gene Expression ,Lymphoproliferative disorders ,CD8-Positive T-Lymphocytes ,Biology ,medicine.disease_cause ,Immunotherapy, Adoptive ,Biochemistry ,Natural killer cell ,Viral Matrix Proteins ,Interleukin 21 ,Cell Line, Tumor ,hemic and lymphatic diseases ,otorhinolaryngologic diseases ,medicine ,Humans ,RNA, Messenger ,DNA Primers ,Base Sequence ,Cell Biology ,Hematology ,Immunotherapy ,Natural killer T cell ,medicine.disease ,Virology ,Epstein–Barr virus ,Killer Cells, Natural ,Leukemia, Large Granular Lymphocytic ,Lymphoma, Extranodal NK-T-Cell ,stomatognathic diseases ,medicine.anatomical_structure ,Cancer research ,RNA, Viral ,CD8 - Abstract
Therapeutic targeting of virus-encoded proteins using cellular immunotherapy has proved successful for Epstein-Barr virus (EBV)–associated posttransplant lymphoproliferative disease. However, the more limited repertoire and immunogenicity of EBV-encoded proteins in other malignancies such as Hodgkin lymphoma and extranodal natural killer (NK)/T lymphoma has been more challenging to target. The immunosubdominant latent membrane protein 2 (LMP2) is considered the optimal target in such Latency II tumors, although data relating to its expression in T/NK malignancies are limited. In addressing the validity of LMP2 as an immunotherapeutic target we found that LMP2-specific effector CD8+ T cells recognized and killed EBV-positive NK- and T-cell tumor lines, despite an apparent absence of LMP2A protein and barely detectable levels of LMP2 transcripts from the conventional LMP2A and LMP2B promoters. We resolved this paradox by identifying in these lines a novel LMP2 mRNA, initiated from within the EBV terminal repeats and containing downstream, epitope-encoding exons. This same mRNA was also highly expressed in primary (extra-nodal) NK/T lymphoma tissue, with virtually undetectable levels of conventional LMP2A/B transcripts. Expression of this novel transcript in T/NK-cell lymphoproliferative diseases validates LMP2 as an attractive target for cellular immunotherapy and implicates this truncated LMP2 protein in NK- and T-cell lymphomagenesis. This study is registered at clinicaltrials.gov as NCT00062868.
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- 2010
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39. Burkitt's lymphoma: The Rosetta Stone deciphering Epstein-Barr virus biology
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Alan B. Rickinson, Gemma L. Kelly, Martin Rowe, and Andrew I. Bell
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Cancer Research ,Apoptosis ,Review ,Biology ,medicine.disease_cause ,Virus ,Cancer pathogenesis ,03 medical and health sciences ,0302 clinical medicine ,c-myc ,immune system diseases ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,Epstein-Barr virus ,Epstein–Barr virus infection ,030304 developmental biology ,0303 health sciences ,Burkitt's lymphoma ,medicine.disease ,Virology ,Epstein–Barr virus ,Burkitt Lymphoma ,3. Good health ,Lymphoma ,Nasopharyngeal carcinoma ,030220 oncology & carcinogenesis ,Human cancer - Abstract
Epstein-Barr virus was originally identified in the tumour cells of a Burkitt's lymphoma, and was the first virus to be associated with the pathogenesis of a human cancer. Studies on the relationship of EBV with Burkitt's lymphoma have revealed important general principles that are relevant to other virus-associated cancers. In addition, the impact of such studies on the knowledge of EBV biology has been enormous. Here, we review some of the key historical observations arising from studies on Burkitt's lymphoma that have informed our understanding of EBV, and we summarise the current hypotheses regarding the role of EBV in the pathogenesis of Burkitt's lymphoma.
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- 2009
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40. Epstein-Barr virus colonization of tonsillar and peripheral blood B-cell subsets in primary infection and persistence
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Gerald Niedobitek, Alan B. Rickinson, Emily Heath, Michael Kuo, Andrew I. Bell, Maike Buettner, W. Bergler, and Sridhar Chaganti
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Adult ,Gene Expression Regulation, Viral ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Palatine Tonsil ,Immunology ,B-Lymphocyte Subsets ,Biology ,CD38 ,medicine.disease_cause ,Biochemistry ,Immunoglobulin D ,Virus ,Antigen ,hemic and lymphatic diseases ,medicine ,Humans ,B cell ,Blood Cells ,Membrane Glycoproteins ,Models, Immunological ,Germinal center ,Cell Biology ,Hematology ,Viral Load ,ADP-ribosyl Cyclase 1 ,Immunoglobulin Class Switching ,Epstein–Barr virus ,Virology ,medicine.anatomical_structure ,Immunoglobulin class switching ,biology.protein ,Immunologic Memory ,Virus Physiological Phenomena - Abstract
Epstein-Barr virus (EBV) persists in the immune host by preferentially colonizing the isotype-switched (IgD−CD27+) memory B-cell pool. In one scenario, this is achieved through virus infection of naive (IgD+CD27−) B cells and their differentiation into memory via germinal center (GC) transit; in another, EBV avoids GC transit and infects memory B cells directly. We report 2 findings consistent with this latter view. First, we examined circulating non–isotype-switched (IgD+CD27+) memory cells, a population that much evidence suggests is GC-independent in origin. Whereas isotype-switched memory had the highest viral loads by quantitative polymerase chain reaction, EBV was detectable in the nonswitched memory pool both in infectious mononucleosis (IM) patients undergoing primary infection and in most long-term virus carriers. Second, we examined colonization by EBV of B-cell subsets sorted from a unique collection of IM tonsillar cell suspensions. Here viral loads were concentrated in B cells with the CD38 marker of GC origin but lacking other GC markers CD10 and CD77. These findings, supported by histologic evidence, suggest that EBV infection in IM tonsils involves extrafollicular B cells expressing CD38 as an activation antigen and not as a marker of ectopic GC activity.
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- 2009
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41. Specific Targeting of the EBV Lytic Phase Protein BNLF2a to the Transporter Associated with Antigen Processing Results in Impairment of HLA Class I-Restricted Antigen Presentation
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Maaike E. Ressing, Malgorzata A. Garstka, Nathan P. Croft, Daniëlle Horst, Emmanuel J. H. J. Wiertz, Andrew D. Hislop, Elisabeth Kremmer, Daphne van Leeuwen, and Alan B. Rickinson
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Immunoprecipitation ,Blotting, Western ,Molecular Sequence Data ,Immunology ,Antigen presentation ,Fluorescent Antibody Technique ,Human leukocyte antigen ,Biology ,Cell Line ,Viral Proteins ,Tapasin ,Transduction, Genetic ,Animals ,Humans ,Immunology and Allergy ,Cytotoxic T cell ,Amino Acid Sequence ,Antigen Presentation ,Reverse Transcriptase Polymerase Chain Reaction ,Histocompatibility Antigens Class I ,Transporter associated with antigen processing ,Flow Cytometry ,Virology ,Lytic cycle ,biology.protein ,TAP2 ,ATP-Binding Cassette Transporters - Abstract
EBV persists for life in the human host while facing vigorous antiviral responses that are induced upon primary infection. This persistence supports the idea that herpesviruses have acquired dedicated functions to avoid immune elimination. The recently identified EBV gene product BNLF2a blocks TAP. As a result, reduced amounts of peptides are transported by TAP from the cytoplasm into the endoplasmic reticulum (ER) lumen for binding to newly synthesized HLA class I molecules. Thus, BNLF2a perturbs detection by cytotoxic T cells. The 60-aa-long BNLF2a protein prevents the binding of both peptides and ATP to TAP, yet further mechanistic insight is, to date, lacking. In this study, we report that EBV BNLF2a represents a membrane-associated protein that colocalizes with its target TAP in subcellular compartments, primarily the ER. In cells devoid of TAP, expression levels of BNLF2a protein are greatly diminished, while ER localization of the remaining BNLF2a is retained. For interactions of BNLF2a with the HLA class I peptide-loading complex, the presence of TAP2 is essential, whereas tapasin is dispensible. Importantly, we now show that in B cells supporting EBV lytic replication, the BNLF2a protein is expressed early in infection, colocalizing and associating with the peptide-loading complex. These results imply that, during productive EBV infection, BNLF2a contributes to TAP inhibition and surface HLA class I down-regulation. In this way, EBV BNLF2a-mediated evasion from HLA class I-restricted T cell immunity contributes to creating a window for undetected virus production.
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- 2009
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42. Hepatitis C virus association with peripheral blood B lymphocytes potentiates viral infection of liver-derived hepatoma cells
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Alan B. Rickinson, J Shaw, John R. Gordon, David H. Adams, Claire Shannon-Lowe, David Mutimer, Peter Balfe, Jane A. McKeating, and Zania Stamataki
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Carcinoma, Hepatocellular ,Hepatitis C virus ,Blotting, Western ,Immunology ,Viral transformation ,Hepacivirus ,medicine.disease_cause ,Biochemistry ,Virus ,medicine ,Animals ,Humans ,Antibody-dependent enhancement ,Immunobiology ,B-Lymphocytes ,CD40 ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Liver Neoplasms ,Hepatitis C ,Cell Biology ,Hematology ,Flow Cytometry ,medicine.disease ,Virology ,Molecular biology ,digestive system diseases ,NS2-3 protease ,biology.protein ,Receptors, Virus ,Antibody - Abstract
Hepatitis C virus (HCV) primarily replicates within the liver, leading to hepatitis, fibrosis, and hepatocellular carcinoma. Infection is also associated with B-cell abnormalities, suggesting an association of the virus with B cells. The infectious JFH-1 strain of HCV can bind primary and immortalized B cells but fails to establish productive infection. However, B cell–associated virus readily infects hepatoma cells, showing an enhanced infectivity compared with extracellular virus. B cells express the viral receptors CD81, SR-BI, and the C-type lectins DC-SIGN and L-SIGN. Antibodies specific for SR-BI and DC-SIGN/L-SIGN reduced B-cell transinfection, supporting a role for these molecules in B-cell association with HCV. Stimulation of B cells with CD40 ligand and interleukin-4 promoted their ability to transinfect hepatoma cells. B cell–associated virus is resistant to trypsin proteolysis and HCV-specific neutralizing antibodies, consistent with particle internalization. HCV promoted the adhesion of primary B cells to Huh-7 hepatomas, providing a mechanism for B-cell retention in the infected liver. In summary, B cells may provide a vehicle for HCV to persist and transmit to the liver.
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- 2009
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43. The Effects of Acute Malaria on Epstein-Barr Virus (EBV) Load and EBV-Specific T Cell Immunity in Gambian Children
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Ramou Njie, Alan B. Rickinson, Sridhar Chaganti, Andrew D. Hislop, Debbie Croom-Carter, Andrew I. Bell, Hilton Whittle, and Hui Jia
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Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Adolescent ,Mononucleosis ,T-Lymphocytes ,T cell ,Population ,Genome, Viral ,CD8-Positive T-Lymphocytes ,Biology ,medicine.disease_cause ,Polymerase Chain Reaction ,Virus ,Open Reading Frames ,Reference Values ,hemic and lymphatic diseases ,parasitic diseases ,medicine ,Humans ,Immunology and Allergy ,Child ,education ,education.field_of_study ,Plasmodium falciparum ,Viral Load ,medicine.disease ,biology.organism_classification ,Epstein–Barr virus ,Virology ,Malaria ,Infectious Diseases ,medicine.anatomical_structure ,Child, Preschool ,Acute Disease ,Immunology ,Gambia ,Viral load - Abstract
Background. To investigate how intense Plasmodium falciparum infection predisposes to Epstein-Barr virus (EBV)-positive Burkitt lymphoma (BL) we analyzed the effect of acute malaria on existing EBV-host balance. Methods. EBV genome loads in peripheral blood mononuclear cells were assayed by quantitative polymerase chain reaction and EBV-specific CD8_ T cell responses were assayed by interferon-y enzyme-linked immunospot assay. Results. Gambian children from whom samples were obtained during an acute malaria attack and again up to 6 weeks later had extremely high viral loads reaching levels that in the United Kingdom are seen only in patients with infectious mononucleosis. Gambian control subjects (children and adults with no recent history of malaria) had lower median viral loads although they were still >10-fold above the median for healthy UK adults. Limited experiments with EBV epitope peptides (restricted through the HLA-B*3501 and HLA-B*5301 alleles) also suggested an impairment of virus-specific CD8+ T cell function in children with malaria but only during acute disease. Conclusions. Acute malaria is associated with sustained increase in EBV load and possibly a transient decrease in EBV-specific T cell surveillance. We infer that the unusually high set point of virus carriage in P. falciparum-challenged populations allied with the parasites capacity to act as a chronic B cell stimulus probably contributes to the pathogenesis of endemic BL.
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- 2009
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44. EBV Latent Membrane Proteins (LMPs) 1 and 2 as Immunotherapeutic Targets: LMP-Specific CD4+ Cytotoxic T Cell Recognition of EBV-Transformed B Cell Lines
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Tracey A. Haigh, Xiaorong Lin, Alan B. Rickinson, Hui Jia, Graham S. Taylor, Edwin P. Hui, and Anthony T.C. Chan
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CD4-Positive T-Lymphocytes ,Epstein-Barr Virus Infections ,T cell ,Molecular Sequence Data ,Immunology ,Human leukocyte antigen ,Biology ,Epitope ,Viral Matrix Proteins ,Epitopes ,Cell Line, Tumor ,Cyclosporin a ,Leukemia, B-Cell ,otorhinolaryngologic diseases ,medicine ,Humans ,Immunology and Allergy ,Cytotoxic T cell ,Amino Acid Sequence ,B cell ,B-Lymphocytes ,Cell Transformation, Viral ,Molecular biology ,Clone Cells ,medicine.anatomical_structure ,Membrane protein ,Cytokines ,Peptides ,CD8 ,T-Lymphocytes, Cytotoxic - Abstract
The EBV-latent membrane proteins (LMPs) 1 and 2 are among only three viral proteins expressed in EBV-associated Hodgkin’s lymphoma and nasopharyngeal carcinoma. Since these tumors are HLA class I and class II-positive, the LMPs could serve as both CD8+ and CD4+ T cell targets. In contrast to CD8 responses, very little is known about CD4 responses to LMPs. In this study, we describe CD4+ T cell clones defining four LMP1- and three LMP2-derived peptide epitopes and their restricting alleles. All clones produced Th1-like cytokines in response to peptide and most killed peptide-loaded target cells by perforin-mediated lysis. Although clones to different epitopes showed different functional avidities in peptide titration assays, avidity per se was a poor predictor of the ability to recognize naturally infected B lymphoblastoid cell lines (LCLs) expressing LMPs at physiologic levels. Some epitopes, particularly within LMP1, consistently mediated strong LCL recognition detectable in cytokine release, cytotoxicity, and outgrowth inhibition assays. Using cyclosporin A to selectively block cytokine release, we found that CD4+ T cell cytotoxicity is the key effector of LCL outgrowth control. We therefore infer that cytotoxic CD4+ T cells to a subset of LMP epitopes could have therapeutic potential against LMP-expressing tumors.
- Published
- 2008
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45. Cancer Virus : The Discovery of the Epstein-Barr Virus
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Dorothy H. Crawford, Ingólfur Johannessen, Alan B. Rickinson, Dorothy H. Crawford, Ingólfur Johannessen, and Alan B. Rickinson
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- Cancer--Etiology, Epstein-Barr virus, Epstein-Barr virus diseases, Herpesvirus 4, Human, Neoplasms--etiology, Epstein-Barr Virus Infections
- Abstract
The Epstein-Barr virus (EBV) was discovered in 1964. At the time, the very idea of a virus underlying a cancer was revolutionary. Cancer is, after all, not catching. Even now, the idea of a virus causing cancer surprises many people. But Epstein-Barr, named after its discoverers, Sir Anthony Epstein and Dr Yvonne Barr, is fascinating for other reasons too. Almost everyone carries it, yet it is only under certain circumstances that it produces disease. It has been associated with different, apparently unrelated, diseases in different populations: Burkitt's Lymphoma, producing tumours in the jaw, in African children; a nasal tumour in China; glandular fever in Europe and the USA; and the majority of cases of Hodgkin's Disease everywhere. This book tells the story of the discovery of the virus, and the recognition of its connection with these various diseases - an account that spans the world and involves some remarkable characters and individual stories.
- Published
- 2014
46. CD4 and CD8 T cell responses to tumour-associated Epstein–Barr virus antigens in nasopharyngeal carcinoma patients
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Nancy H. Gudgeon, C. Kit Lin, Xiaorong Lin, Alan B. Rickinson, Martin Barnardo, Xue Qun, Hui Jia, Anthony T.C. Chan, Edwin P. Hui, and Graham S. Taylor
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Adult ,CD4-Positive T-Lymphocytes ,Herpesvirus 4, Human ,Cancer Research ,T cell ,Immunology ,Epitopes, T-Lymphocyte ,Human leukocyte antigen ,CD8-Positive T-Lymphocytes ,Biology ,medicine.disease_cause ,Epitope ,Viral Matrix Proteins ,Antigen ,otorhinolaryngologic diseases ,medicine ,Humans ,Immunology and Allergy ,Cytotoxic T cell ,Antigens, Viral ,Nasopharyngeal Neoplasms ,Middle Aged ,medicine.disease ,Epstein–Barr virus ,stomatognathic diseases ,medicine.anatomical_structure ,Epstein-Barr Virus Nuclear Antigens ,Oncology ,Nasopharyngeal carcinoma ,Cancer research ,CD8 - Abstract
Nasopharyngeal carcinoma (NPC), an Epstein-Barr virus (EBV)-associated tumour common in Southern Chinese populations, is a potentially important target for T cell-based immunotherapy. The tumour cells are HLA class I- and II-positive and express a limited subset of EBV latent proteins, namely the nuclear antigen EBNA1 and the latent membrane proteins LMP2 and (in some cases) LMP1. To ask whether the tumour develops in the presence of a potentially protective host response or in its absence, we set out to determine the prevailing levels of CD4+ and CD8+ T cell memory to these proteins in NPC patients at tumour diagnosis. We first screened healthy Chinese donors against Chinese strain EBNA1, LMP1 and LMP2 sequences in Elispot assays of interferon-gamma release and identified the immunodominant CD4+ and CD8+ epitope peptides presented by common Chinese HLA alleles. Then, comparing 60 patients with70 healthy controls on peptide epitope mini-panels, we found that T cell memory to CD4 epitopes in all three proteins was unimpaired in the blood of patients at diagnosis. In most cases NPC patients also showed detectable responses to CD8 epitopes relevant to their HLA type, the one consistent exception being the absence in patients of a B*4001-restricted response to LMP2. We infer that NPC arises in patients whose prevailing levels of T cell memory to tumour-associated EBV proteins is largely intact; the therapeutic goal must therefore be to re-direct the existing memory repertoire more effectively against antigen-expressing tumour cells.
- Published
- 2007
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47. Epstein-Barr Virus Exploits BSAP/Pax5 To Achieve the B-Cell Specificity of Its Growth-Transforming Program
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Alan B. Rickinson, Wolfgang Hammerschmidt, Rosemary J. Tierney, Isabel A. Hutchings, Andrew I. Bell, Claire Shannon-Lowe, Markus Altmann, and Jasdeep Nagra
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Gene Expression Regulation, Viral ,Herpesvirus 4, Human ,Transcription, Genetic ,Immunology ,Mutant ,Genome, Viral ,Biology ,medicine.disease_cause ,Microbiology ,Herpesviridae ,Virus ,Transcription (biology) ,Virology ,medicine ,Humans ,Viral Regulatory and Accessory Proteins ,Promoter Regions, Genetic ,Cells, Cultured ,B cell ,B-Lymphocytes ,Binding Sites ,Activator (genetics) ,PAX5 Transcription Factor ,Wild type ,Epithelial Cells ,Cell Transformation, Viral ,Molecular biology ,Epstein–Barr virus ,Genome Replication and Regulation of Viral Gene Expression ,medicine.anatomical_structure ,Organ Specificity ,Insect Science ,Gene Deletion - Abstract
Epstein-Barr virus (EBV) can infect various cell types but limits its classical growth-transforming function to B lymphocytes, the cells in which it persists in vivo. Transformation initiates with the activation of Wp, a promoter present as tandemly repeated copies in the viral genome. Assays with short Wp reporter constructs have identified two promoter-activating regions, one of which (UAS2) appears to be lineage independent, while the other (UAS1) was B-cell specific and contained two putative binding sites for the B-cell-specific activator protein BSAP/Pax5. To address the physiologic relevance of these findings, we first used chromosome immunoprecipitation assays and found that BSAP is indeed bound to Wp sequences on the EBV genome in transformed cells. Thereafter, we constructed recombinant EBVs carrying two Wp copies, both wild type, with UAS1 or UAS2 deleted, or mutated in the BSAP binding sites. All the viruses delivered their genomes to the B-cell nucleus equally well. However, the BSAP binding mutant (and the virus with UAS1 deleted) showed no detectable activity in B cells, whether measured by early Wp transcription, expression of EBV latent proteins, or outgrowth of transformed cells. This was a B-cell-specific defect since, on entry into epithelial cells, an environment where Wp is not the latent promoter of choice, all the Wp mutant viruses initiated infection as efficiently as wild-type virus. We infer that EBV ensures the B-cell specificity of its growth-transforming function by exploiting BSAP/Pax5 as a lineage-specific activator of the transforming program.
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- 2007
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48. A CD8+ T cell immune evasion protein specific to Epstein-Barr virus and its close relatives in Old World primates
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Maaike E. Ressing, Daphne van Leeuwen, Alan B. Rickinson, Victoria Anne Pudney, Andrew D. Hislop, Danijela Koppers-Lalic, Nathan P. Croft, Daniëlle Horst, Jacques Neefjes, and Emmanuel J. H. J. Wiertz
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Herpesvirus 4, Human ,T cell ,Molecular Sequence Data ,Immunology ,Human leukocyte antigen ,CD8-Positive T-Lymphocytes ,Major histocompatibility complex ,medicine.disease_cause ,Article ,03 medical and health sciences ,0302 clinical medicine ,HLA Antigens ,medicine ,Animals ,Humans ,Immunology and Allergy ,Cytotoxic T cell ,Amino Acid Sequence ,Cloning, Molecular ,Lytic Phase ,Herpesviridae ,030304 developmental biology ,Genetics ,0303 health sciences ,Sequence Homology, Amino Acid ,biology ,Cercopithecidae ,Articles ,Flow Cytometry ,Epstein–Barr virus ,Virology ,medicine.anatomical_structure ,Gene Expression Regulation ,Lytic cycle ,Immune System ,biology.protein ,Peptides ,CD8 ,030215 immunology - Abstract
gamma 1-Herpesviruses such as Epstein-Barr virus (EBV) have a unique ability to amplify virus loads in vivo through latent growth-transforming infection. Whether they, like alpha- and beta-herpesviruses, have been driven to actively evade immune detection of replicative (lytic) infection remains a moot point. We were prompted to readdress this question by recent work (Pudney, V.A., A.M. Leese, A.B. Rickinson, and A.D. Hislop. 2005. J. Exp. Med. 201:349-360; Ressing, M.E., S.E. Keating, D. van Leeuwen, D. Koppers-Lalic, I.Y. Pappworth, E.J.H.J. Wiertz, and M. Rowe. 2005. J. Immunol. 174:6829-6838) showing that, as EBV-infected cells move through the lytic cycle, their susceptibility to EBV-specific CD8(+) T cell recognition falls dramatically, concomitant with a reductions in transporter associated with antigen processing (TAP) function and surface human histocompatibility leukocyte antigen (HLA) class I expression. Screening of genes that are unique to EBV and closely related gamma 1-herpesviruses of Old World primates identified an early EBV lytic cycle gene, BNLF2a, which efficiently blocks antigen-specific CD8(+) T cell recognition through HLA-A-, HLA-B-, and HLA-C-restricting alleles when expressed in target cells in vitro. The small (60-amino acid) BNLF2a protein mediated its effects through interacting with the TAP complex and inhibiting both its peptide- and ATP-binding functions. Furthermore, this targeting of the major histocompatibility complex class I pathway appears to be conserved among the BNLF2a homologues of Old World primate gamma 1-herpesviruses. Thus, even the acquisition of latent cycle genes endowing unique growth-transforming ability has not liberated these agents from evolutionary pressure to evade CD8(+) T cell control over virus replicative foci.
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- 2007
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49. Genome diversity of Epstein-Barr virus from multiple tumor types and normal infection
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Alan B. Rickinson, Simon J. Watson, Samantha Correia, Martin J. Allday, Craig Corton, Anne L. Palser, Mohammed M. Ba Abdullah, Paul J. Farrell, Lawrence S. Young, Paul Kellam, Paul Murray, John R. Arrand, Nicholas E. Grayson, Robert E. White, Matthew Cotten, Imperial College Trust, and King Abdulaziz City for Science and Technology (KA
- Subjects
SELECTION ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Epitopes, T-Lymphocyte ,medicine.disease_cause ,Genome ,hemic and lymphatic diseases ,PHYLOGENIES ,Antigens, Viral ,11 Medical and Health Sciences ,Phylogeny ,Genetics ,Recombination, Genetic ,education.field_of_study ,ALGORITHMS ,3. Good health ,AMINO-ACID ,Carrier State ,Life Sciences & Biomedicine ,Immunology ,Population ,BIOLOGY ,Single-nucleotide polymorphism ,Genome, Viral ,Biology ,Microbiology ,INTERTYPIC RECOMBINANTS ,SEQUENCE ,Polymorphism, Single Nucleotide ,Virus ,Viral Matrix Proteins ,Virology ,07 Agricultural and Veterinary Sciences ,Cell Line, Tumor ,Genetic variation ,medicine ,Humans ,Amino Acid Sequence ,education ,Gene ,QR355 ,CHINESE POPULATION ,TOOLS ,Science & Technology ,STRAINS ,Genetic Variation ,LYMPHOMAGENESIS ,06 Biological Sciences ,Epstein–Barr virus ,Human genetics ,INDIVIDUALS ,Genetic Diversity and Evolution ,Epstein-Barr Virus Nuclear Antigens ,Insect Science ,DNA, Viral - Abstract
Epstein-Barr virus (EBV) infects most of the world's population and is causally associated with several human cancers, but little is known about how EBV genetic variation might influence infection or EBV-associated disease. There are currently no published wild-type EBV genome sequences from a healthy individual and very few genomes from EBV-associated diseases. We have sequenced 71 geographically distinct EBV strains from cell lines, multiple types of primary tumor, and blood samples and the first EBV genome from the saliva of a healthy carrier. We show that the established genome map of EBV accurately represents all strains sequenced, but novel deletions are present in a few isolates. We have increased the number of type 2 EBV genomes sequenced from one to 12 and establish that the type 1/type 2 classification is a major feature of EBV genome variation, defined almost exclusively by variation of EBNA2 and EBNA3 genes, but geographic variation is also present. Single nucleotide polymorphism (SNP) density varies substantially across all known open reading frames and is highest in latency-associated genes. Some T-cell epitope sequences in EBNA3 genes show extensive variation across strains, and we identify codons under positive selection, both important considerations for the development of vaccines and T-cell therapy. We also provide new evidence for recombination between strains, which provides a further mechanism for the generation of diversity. Our results provide the first global view of EBV sequence variation and demonstrate an effective method for sequencing large numbers of genomes to further understand the genetics of EBV infection. IMPORTANCE Most people in the world are infected by Epstein-Barr virus (EBV), and it causes several human diseases, which occur at very different rates in different parts of the world and are linked to host immune system variation. Natural variation in EBV DNA sequence may be important for normal infection and for causing disease. Here we used rapid, cost-effective sequencing to determine 71 new EBV sequences from different sample types and locations worldwide. We showed geographic variation in EBV genomes and identified the most variable parts of the genome. We identified protein sequences that seem to have been selected by the host immune system and detected variability in known immune epitopes. This gives the first overview of EBV genome variation, important for designing vaccines and immune therapy for EBV, and provides techniques to investigate relationships between viral sequence variation and EBV-associated diseases.
- Published
- 2015
50. Primary immunodeficiencies and the control of Epstein-Barr virus infection
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Alan B. Rickinson and Umaimainthan Palendira
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Herpesvirus 4, Human ,Mononucleosis ,T-Lymphocytes ,Disease ,Biology ,medicine.disease_cause ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Immune system ,History and Philosophy of Science ,Immunopathology ,medicine ,Animals ,Humans ,Infectious Mononucleosis ,Epstein–Barr virus infection ,Immunity, Cellular ,General Neuroscience ,Genetic Diseases, Inborn ,Immunologic Deficiency Syndromes ,medicine.disease ,Epstein–Barr virus ,Virology ,Burkitt Lymphoma ,Killer Cells, Natural ,Immunology ,Primary immunodeficiency - Abstract
Human primary immunodeficiency (PID) states, where mutations in single immune system genes predispose individuals to certain infectious agents and not others, are experiments of nature that hold important lessons for the immunologist. The number of genetically defined PIDs is rising rapidly, as is the opportunity to learn from them. Epstein-Barr virus (EBV), a human herpesvirus, has long been of interest because of its complex interaction with the immune system. Thus, it causes both infectious mononucleosis (IM), an immunopathologic disease associated with exaggerated host responses, and at least one malignancy, EBV-positive lymphoproliferative disease, when those responses are impaired. Here, we describe the full range of PIDs currently linked with an increased risk of EBV-associated disease. These provide examples where IM-like immunopathology is fatally exaggerated, and others where responses impaired at the stage of induction, expansion, or effector function predispose to malignancy. Current evidence from this rapidly moving field supports the view that lesions in both natural killer cell and T cell function can lead to EBV pathology.
- Published
- 2015
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