• Epstein–Barr virus
• HIV
• gammaherpesvirus
• human herpesvirus-4
• human herpesvirus-8
• rhadinovirus
• ubiquitin

• Humans
• Ubiquitin-Protein Ligases/metabolism
• HIV/metabolism
• Epstein-Barr Virus Infections
• Herpesvirus 4, Human/metabolism
• Gammaherpesvirinae/metabolism
• Herpesvirus 8, Human/genetics
• Herpesvirus 8, Human/metabolism
• Ubiquitin/metabolism
• HIV Infections
• Virus Replication/physiology

• R15 AI157907 NIAID NIH HHS
• T32 GM146694 NIGMS NIH HHS

• Venezuelan equine encephalitis virus
• alphavirus
• early growth response protein 1
• herpesvirus
• inflammation
• transcription
• virus

• Defense Threat Reduction Agency (DTRA)

Interactions involving Epstein–Barr virus (EBV) LMP2A and Nedd4 family E3 ubiquitin–protein ligases promote the ubiquitination of LMP2A-associated proteins, which results in the perturbation of normal B-cell signaling. Here, we solved the solution structure of the WW2 domain of hAIP4 and investigated the binding mode involving the N-terminal domain of LMP2A and the WW2 domain. The WW2 domain presented a conserved WW domain scaffold with a three-stranded anti-parallel β-sheet and bound two PY motifs via different binding mechanisms. Our NMR titration and ITC data demonstrated that the PY motifs of LMP2A can recognize and interact weakly with the XP groove of the WW2 domain (residues located around the third β-strand), and then residues between two PY motifs optimize the binding by interacting with the loop 1 region of the WW2 domain. In particular, the residue Val15 in the hairpin loop region between β1 and β2 of the WW2 domain exhibited unique changes depending on the terminal residues of the PY motif. This result suggested that the hairpin loop is responsible for additional interactions outside the XP groove, and this hypothesis was confirmed in a deuterium exchange experiment. These weak but wide interactions can stabilize the complex formed between the PY and WW domains.

• AIP4
• E3 ubiquitin ligase
• EBV
• LMP2A
• NMR

• Antiviral signaling
• Calcium
• Calcium homeostasis
• Mitochondria
• Mitochondrial calcium uniporter
• Virus

• haematological cancer
• cell signalling

• Epstein-Barr virus
• dexamethasone
• epithelial-mesenchymal transition
• pulmonary fibrosis
• respiratory syncytial virus

• Humans
• Epithelial-Mesenchymal Transition/drug effects
• A549 Cells
• Herpesvirus 4, Human/drug effects
• Herpesvirus 4, Human/physiology
• Herpesvirus 4, Human/genetics
• Dexamethasone/pharmacology
• Cytokines/metabolism
• Epithelial Cells/virology
• Epithelial Cells/drug effects
• Cadherins/metabolism
• Viral Matrix Proteins/metabolism
• Viral Matrix Proteins/genetics
• Vimentin/metabolism
• Vimentin/genetics
• Epstein-Barr Virus Infections/virology
• Actins/metabolism

• B cell receptor
• Epstein–Barr virus
• lymphoma
• signal transduction

Due to its high morbidity and mortality, gastric cancer is a topic of a great concern throughout the world. Major ways of treatment are gastrectomy and chemotherapy, unfortunately they are not always successful. In a search for more efficient therapy strategies, viruses and their potential seem to be an important issue. On one hand, several oncogenic viruses have been noticed in the case of gastric cancer, making the positive treatment even more advantageous, but on the other, viruses exist with a potential therapeutic role in this malignancy.

• cancer treatment
• gastric cancer
• immunology
• oncogenic viruses
• oncolytic viruses

• Epstein Barr virus
• epithelial cancers
• latency program
• lymphomas

• Wellcome Trust

• EBV (Epstein-Barr virus)
• EBV latency
• EBV-associated cancers
• Lymphoproliferative disease (LPD)
• viral life cycle inhibitors

• L30 CA189070 NCI NIH HHS

The Epstein-Barr virus (EBV) oncoproteins latent membrane protein 1 (LMP1) and LMP2A constitutively activate multiple signaling pathways, and both have been shown to interact with cellular ubiquitin ligases and affect cellular ubiquitination. To detect the LMP1- and LMP2A-mediated effects on the global cellular proteome, epithelial cell lines expressing LMP1 or LMP2A were analyzed using label-free quantitative proteomics. To identify proteins whose ubiquitination is affected by the viral proteins, the cells were cultured in the presence and absence of deubiquitinase (DUB) and proteasome inhibitors. More than 7,700 proteins were identified with high confidence and considerably more proteins showed significant differences in expression in the presence of inhibitors. Few of the differentially expressed proteins with or without inhibitors were common between LMP1 and LMP2A, confirming that the viral proteins induce unique changes in cell expression and function. However, ingenuity pathway analysis (IPA) of the data indicated that LMP1 and LMP2A modulate many of the same cellular regulatory pathways, including cell death and survival, cell movement, and actin filament dynamics. In addition, various proteasome subunits, ubiquitin-specific peptidases and conjugating enzymes, vesicle trafficking proteins, and NF-κB and mitogen-activated protein kinase signaling proteins were affected by LMP1 or LMP2A. These findings suggest that LMP1 and LMP2A may commonly target critical cell pathways through effects on distinct genes, with many cellular proteins modified by ubiquitination and/or degradation.

The Epstein-Barr virus proteins latent membrane protein 1 and 2 have potent effects on cell growth and signaling. Both proteins bind to specific ubiquitin ligases and likely modulate the cellular proteome through ubiquitin-mediated effects on stability and intracellular location. In this study, a comprehensive proteomic analysis of the effects of LMP1 and LMP2A revealed that both proteins affected proteasome subunits, ubiquitin-specific conjugases and peptidases, and vesical trafficking proteins. The data suggest that the effects of these proteins on the abundance and ubiquitination of cellular proteins are in part responsible for their effects on cell growth regulation.

• Epstein-Barr virus associated gastric carcinoma
• dihydroartemisinin
• latent membrane protein 2A
• proliferation

Lytic reactivation of EBV has been reported to play an important role in human diseases, including NPC carcinogenesis. Inhibition of EBV reactivation is considered to be of great benefit in the treatment of virus-associated diseases. For this purpose, we screened for inhibitory compounds and found that apigenin, a flavonoid, seemed to have the ability to inhibit EBV reactivation.

We performed western blotting, immunofluorescence and luciferase analyses to determine whether apigenin has anti-EBV activity.

Apigenin inhibited expression of the EBV lytic proteins, Zta, Rta, EAD and DNase in epithelial and B cells. It also reduced the number of EBV-reactivating cells detectable by immunofluorescence analysis. In addition, apigenin has been found to reduce dramatically the production of EBV virions. Luciferase reporter analysis was performed to determine the mechanism by which apigenin inhibits EBV reactivation: apigenin suppressed the activity of the immediate-early (IE) gene Zta and Rta promoters, suggesting it can block initiation of the EBV lytic cycle.

Taken together, apigenin inhibits EBV reactivation by suppressing the promoter activities of two viral IE genes, suggesting apigenin is a potential dietary compound for prevention of EBV reactivation.

The online version of this article (doi:10.1186/s12929-016-0313-9) contains supplementary material, which is available to authorized users.

• Apigenin
• Epstein-Barr virus
• Nasopharyngeal carcinoma
• Reactivation

• Epstein-Barr virus (EBV)
• Kaposi’s sarcoma-associated herpesvirus (KSHV)
• cell membrane remodeling

• Animals
• Cell Membrane/virology
• Epstein-Barr Virus Infections/virology
• Herpesviridae Infections/virology
• Herpesvirus 4, Human/genetics
• Herpesvirus 4, Human/physiology
• Herpesvirus 8, Human/genetics
• Herpesvirus 8, Human/physiology
• Humans
• Virus Replication

• Carcinogenesis
• Gene Expression Regulation
• Herpesvirus 4, Human/physiology
• Host-Pathogen Interactions
• Humans
• Virus Latency

• P01 CA174439 NCI NIH HHS
• U54 CA190158 NCI NIH HHS
• P30 DK050306 NIDDK NIH HHS
• U54CA190158 NCI NIH HHS
• R01 CA177423 NCI NIH HHS
• P30 CA016520 NCI NIH HHS
• P30CA016520 NCI NIH HHS
• P30DK050306 NIDDK NIH HHS
• P01CA174439 NCI NIH HHS

• B cells
• LMP2A
• autoimmune diseases
• germinal center
• plasma cell differentiation

The common pathogen Epstein-Barr virus (EBV) transforms normal human B cells and can cause cancer. Latent membrane protein 2A (LMP2A) of EBV supports activation and proliferation of infected B cells and is expressed in many types of EBV-associated cancer. It is not clear how latent EBV infection and cancer escape elimination by host immunity, and it is unknown whether LMP2A can influence the interaction of EBV-infected cells with the immune system. We infected primary B cells with EBV deleted for LMP2A, and established lymphoblastoid cell lines (LCLs). We found that CD8+ T cell clones showed higher reactivity against LMP2A-deficient LCLs compared to LCLs infected with complete EBV. We identified several potential mediators of this immunomodulatory effect. In the absence of LMP2A, expression of some EBV latent antigens was elevated, and cell surface expression of MHC class I was marginally increased. LMP2A-deficient LCLs produced lower amounts of IL-10, although this did not directly affect CD8+ T cell recognition. Deletion of LMP2A led to several changes in the cell surface immunophenotype of LCLs. Specifically, the agonistic NKG2D ligands MICA and ULBP4 were increased. Blocking experiments showed that NKG2D activation contributed to LCL recognition by CD8+ T cell clones. Our results demonstrate that LMP2A reduces the reactivity of CD8+ T cells against EBV-infected cells, and we identify several relevant mechanisms.

Epstein-Barr virus (EBV) is carried by most humans. It can cause several types of cancer. In healthy infected people, EBV persists for life in a "latent" state in white blood cells called B cells. For infected persons to remain healthy, it is crucial that they harbor CD8-positive "killer" T cells that recognize and destroy precancerous EBV-infected cells. However, this protection is imperfect, because the virus is not eliminated from the body, and the danger of EBV-associated cancer remains. How does the virus counteract CD8+ T cell control? Here we study the effects of latent membrane protein 2A (LMP2A), which is an important viral molecule because it is present in several types of EBV-associated cancers, and in latently infected cells in healthy people. We show that LMP2A counteracts the recognition of EBV-infected B cells by antiviral killer cells. We found a number of mechanisms that are relevant to this effect. Notably, LMP2A disturbs expression of molecules on B cells that interact with NKG2D, a molecule on the surface of CD8+ T cells that aids their activation. In this way, LMP2A weakens important immune responses against EBV. Similar mechanisms may operate in different types of LMP2A-expressing cancers caused by EBV.

The Epstein-Barr virus (EBV) nuclear proteins EBNA3A, EBNA3B, and EBNA3C interact with the cell DNA binding protein RBPJ and regulate cell and viral genes. Repression of the CDKN2A tumor suppressor gene products p16^INK4A and p14^ARF by EBNA3A and EBNA3C is critical for EBV mediated transformation of resting B lymphocytes into immortalized lymphoblastoid cell lines (LCLs). To define the composition of endogenous EBNA3 protein complexes, we generated lymphoblastoid cell lines (LCLs) expressing flag-HA tagged EBNA3A, EBNA3B, or EBNA3C and used tandem affinity purification to isolate each EBNA3 complex. Our results demonstrated that each EBNA3 protein forms a distinct complex with RBPJ. Mass-spectrometry revealed that the EBNA3A and EBNA3B complexes also contained the deubquitylation complex consisting of WDR48, WDR20, and USP46 (or its paralog USP12) and that EBNA3C complexes contained WDR48. Immunoprecipitation confirmed that EBNA3A, EBNA3B, and EBNA3C association with the USP46 complex. Using chromatin immunoprecipitation, we demonstrate that WDR48 and USP46 are recruited to the p14^ARF promoter in an EBNA3C dependent manner. Mapping studies were consistent with WDR48 being the primary mediator of EBNA3 association with the DUB complex. By ChIP assay, WDR48 was recruited to the p14^ARF promoter in an EBNA3C dependent manner. Importantly, WDR48 associated with EBNA3A and EBNA3C domains that are critical for LCL growth, suggesting a role for USP46/USP12 in EBV induced growth transformation.

Epstein-Barr virus (EBV) is a gammaherpesvirus implicated in the pathogenesis of multiple malignancies, including Burkitt lymphoma, Hodgkin lymphoma, post-transplant lymphoproliferative disease (PTLD), nasopharyngeal carcinoma, and gastric carcinoma. EBV infection of resting B-lymphocytes drives them to proliferate as lymphoblastoid cell lines (LCLs), an in vitro model of PTLD. LCLs express a limited EBV gene repertoire, including six nuclear proteins (EBNA1, 2, 3A, 3B, 3C, and LP), three integral membrane proteins (LMP1, 2A, and 2B), and more than 30 micro RNAs. EBNA2 and the EBNA3 proteins are transcription factors that regulate viral and cell gene expression through the cell DNA binding protein RBPJ. In this study, we established LCLs transformed by recombinant EBV genomes in which a Flag-HA epitope tag is fused in-frame to the C-terminus of EBNA3A, EBNA3B or EBNA3C. Using these LCLs, we purified endogenous EBNA3 complexes and identified the USP46 deubiquitinating enzyme (DUB) and its associated chaperones WDR48 and WDR20 as EBNA3 binding proteins. We find that EBNA3s interact primarily with the WDR48 protein and that loss of WDR48 interaction with EBNA3A or EBNA3C impairs LCL growth. This study represents the first characterization of EBNA3 complexes from LCLs and implicates the USP46 DUB complex in EBNA3 mediated gene regulation.

• Blotting, Western
• Cell Line
• Cell Proliferation
• Cell Transformation, Viral/genetics
• Chromatin Immunoprecipitation
• Endopeptidases/genetics
• Endopeptidases/metabolism
• Epstein-Barr Virus Nuclear Antigens/metabolism
• Gene Expression Regulation, Viral/genetics
• Herpesvirus 4, Human/genetics
• Herpesvirus 4, Human/metabolism
• Humans
• Immunoprecipitation
• Mass Spectrometry
• Ubiquitin Thiolesterase/genetics
• Ubiquitin Thiolesterase/metabolism

• R01 CA047006 NCI NIH HHS
• R01 DE023939 NIDCR NIH HHS
• R35 CA047006 NCI NIH HHS
• CA47006 NCI NIH HHS

Latent Epstein–Barr virus (EBV) infection has a substantial role in causing many human disorders. The persistence of these viral genomes in all malignant cells, yet with the expression of limited latent genes, is consistent with the notion that EBV latent genes are important for malignant cell growth. While the EBV-encoded nuclear antigen-1 (EBNA-1) and latent membrane protein-2A (LMP-2A) are critical, the EBNA-leader proteins, EBNA-2, EBNA-3A, EBNA-3C and LMP-1, are individually essential for in vitro transformation of primary B cells to lymphoblastoid cell lines. EBV-encoded RNAs and EBNA-3Bs are dispensable. In this review, the roles of EBV latent genes are summarized.

• Epstein-Barr virus
• LMP2A
• Lyn
• Signal transduction
• Syk

Shigella flexneri interacts with B cells and induces apoptosis via IpaD binding to TLR2.

Antibody-mediated immunity to Shigella, the causative agent of bacillary dysentery, requires several episodes of infection to get primed and is short-lasting, suggesting that the B cell response is functionally impaired. We show that upon ex vivo infection of human colonic tissue, invasive S. flexneri interacts with and occasionally invades B lymphocytes. The induction of a type three secretion apparatus (T3SA)–dependent B cell death is observed in the human CL-01 B cell line in vitro, as well as in mouse B lymphocytes in vivo. In addition to cell death occurring in Shigella-invaded CL-01 B lymphocytes, we provide evidence that the T3SA needle tip protein IpaD can induce cell death in noninvaded cells. IpaD binds to and induces B cell apoptosis via TLR2, a signaling receptor thus far considered to result in activation of B lymphocytes. The presence of bacterial co-signals is required to sensitize B cells to apoptosis and to up-regulate tlr2, thus enhancing IpaD binding. Apoptotic B lymphocytes in contact with Shigella-IpaD are detected in rectal biopsies of infected individuals. This study therefore adds direct B lymphocyte targeting to the diversity of mechanisms used by Shigella to dampen the host immune response.

Epstein-Barr virus (EBV) is a human herpesvirus, which is causally associated with the development of several B lymphocytic malignancies that include Burkitt's lymphomas, Hodgkin's disease, AIDS and posttransplant associated lymphomas. The transforming activity of EBV is orchestrated by several latent viral proteins that mimic and modulate cellular growth promoting and antiapoptotic signaling pathways, which involve among others the activity of protein kinases. In an effort to identify small molecule inhibitors of the growth of EBV-transformed B lymphocytes a library of 254 kinase inhibitors was screened. This effort identified two tyrosine kinase inhibitors and two MEK inhibitors that compromised preferentially the viability of EBV-infected human B lymphocytes. Our findings highlight the possible dependence of EBV-infected B lymphocytes on specific kinase-regulated pathways underlining the potential for the development of small molecule-based therapeutics that could target selectively EBV-associated human B lymphocyte malignancies.

• Ataxia Telangiectasia Mutated Proteins
• Base Sequence
• Cell Cycle Proteins/antagonists & inhibitors
• Cell Cycle Proteins/genetics
• Cell Cycle Proteins/metabolism
• Cell Line
• DNA Primers
• DNA-Binding Proteins/antagonists & inhibitors
• DNA-Binding Proteins/genetics
• DNA-Binding Proteins/metabolism
• Herpesvirus 4, Human/physiology
• Humans
• Imidazoles/metabolism
• Morpholines/pharmacology
• Phosphatidylinositol 3-Kinases/genetics
• Phosphatidylinositol 3-Kinases/metabolism
• Phosphoinositide-3 Kinase Inhibitors
• Piperazines/metabolism
• Protein Serine-Threonine Kinases/antagonists & inhibitors
• Protein Serine-Threonine Kinases/genetics
• Protein Serine-Threonine Kinases/metabolism
• Pyrones/pharmacology
• Reverse Transcriptase Polymerase Chain Reaction
• Tumor Suppressor Protein p53/physiology
• Tumor Suppressor Proteins/antagonists & inhibitors
• Tumor Suppressor Proteins/genetics
• Tumor Suppressor Proteins/metabolism
• Virus Activation

• R01 CA066519 NCI NIH HHS
• R01 CA058853 NCI NIH HHS
• P01-CA022443 NCI NIH HHS
• T32 AI078985 NIAID NIH HHS
• R01-CA66519 NCI NIH HHS
• P01 CA022443 NCI NIH HHS
• R01-CA58853 NCI NIH HHS

Latent membrane protein 1 (LMP1) and LMP2A affect cell growth in both epithelial cells and lymphocytes. In this study, the effects on cellular gene expression were determined by microarray analysis of transgenic mice expressing LMP1, LMP2A, or both using the immunoglobulin heavy chain promoter and enhancer. Large differential changes were detected, indicating that LMP1 and LMP2A can both potently affect host gene transcription, inducing distinct transcriptional profiles. Seventy percent of the changes detected in LMP1/2A doubly transgenic lymphocytes were also modulated by LMP1 or LMP2A alone. These common and unique expression changes indicate that the combined effects of LMP1 and LMP2A may be additive, synergistic, or inhibitory. Using significant pathway analysis, the expression changes detected in LMP1, LMP2A, and LMP1/2A transgenic B lymphocytes were predicted to commonly target cancer and inflammatory pathways. Additionally, using the correlation coefficient to calculate the regulation of known c-Rel and Stat3 transcriptional targets, both were found to be enhanced in LMP1 lymphocytes and lymphomas, and a selection of Stat3 targets was further evaluated and confirmed using quantitative reverse transcription-PCR (RT-PCR). Analyses of the effects on cell growth and viability revealed that LMP2A transgenic lymphocytes had the greatest enhanced viability in vitro; however, doubly transgenic lymphocytes (LMP1/2A) did not have enhanced survival in culture and these mice were similar to negative littermates. These findings indicate that the combined expression of LMP1 and LMP2A has potentially different biological outcomes than when the two proteins are expressed individually.

The Epstein-Barr virus proteins latent membrane protein 1 (LMP1) and LMP2A have potent effects on cell growth. In transgenic mice that express these proteins in B lymphocytes, the cell growth and survival properties are also affected. LMP1 transgenic mice have increased development of lymphoma, and the LMP1 lymphocytes have increased viability in culture. LMP2A transgenic lymphocytes have altered B cell development and enhanced survival. In this study, analysis of the cellular gene expression changes in transgenic LMP1 and LMP2A lymphocytes and LMP1 lymphomas revealed that both transgenes individually and in combination affected pathways important for the development of cancer and inflammation. Importantly, the combined expression of the two proteins had unique effects on cellular expression and cell viability. This is the first study to look at the combined effects of LMP1 and LMP2A on global changes in host gene expression.

Autophagy is an intracellular degradation pathway for long-lived proteins and organelles. This process is activated above basal levels upon cell intrinsic or environmental stress and dysregulation of autophagy has been linked to various human diseases, including those caused by viral infection. Many viruses have evolved strategies to directly interfere with autophagy, presumably to facilitate their replication or to escape immune detection. However, in some cases, modulation of autophagy appears to be a consequence of the virus disturbing the cell’s metabolic signaling networks. Here, we summarize recent advances in research at the interface of autophagy and viral infection, paying special attention to strategies that human tumor viruses have evolved.

• epstein–barr virus
• post-transplant lymphoproliferative disorder
• b cell

• B-Lymphocytes/virology
• Gene Expression Regulation, Viral
• Graft Survival
• Herpesvirus 4, Human/metabolism
• Humans
• Immunosuppressive Agents/pharmacology
• Immunotherapy/methods
• Intracellular Signaling Peptides and Proteins/metabolism
• Lymphoma, B-Cell/virology
• Lymphoproliferative Disorders/therapy
• Lymphoproliferative Disorders/virology
• Models, Biological
• Organ Transplantation/adverse effects
• Phosphatidylinositol 3-Kinases/metabolism
• Postoperative Complications
• Protein-Tyrosine Kinases/metabolism
• Syk Kinase
• Viral Matrix Proteins/metabolism

• R01 AI041769 NIAID NIH HHS
• T32 AI007290 NIAID NIH HHS

The latent membrane protein (LMP) 2A of Epstein-Barr virus (EBV) is expressed during different latency stages of EBV-infected B cells in which it triggers activation of cytoplasmic protein tyrosine kinases. Early studies revealed that an immunoreceptor tyrosine-based activation motif (ITAM) in the cytoplasmic N-terminus of LMP2A can trigger a transient increase of the cytosolic Ca²⁺ concentration similar to that observed in antigen-activated B cells when expressed as a chimeric transmembrane receptor. Even so, LMP2A was subsequently ascribed an inhibitory rather than an activating function because its expression seemed to partially inhibit B cell antigen receptor (BCR) signaling in EBV-transformed B cell lines. However, the analysis of LMP2A signaling has been hampered by the lack of cellular model systems in which LMP2A can be studied without the influence of other EBV-encoded factors.

We have reanalyzed LMP2A signaling using B cells in which LMP2A is expressed in an inducible manner in the absence of any other EBV signaling protein. This allowed us for the first time to monitor LMP2A signaling in statu nascendi as it occurs during the EBV life cycle in vivo. We show that mere expression of LMP2A not only stimulated protein tyrosine kinases but also induced phospholipase C-γ2-mediated Ca²⁺ oscillations followed by activation of the extracellular signal-regulated kinase (Erk) mitogen-activated protein kinase pathway and induction of the lytic EBV gene bzlf1. Furthermore, expression of the constitutively phosphorylated LMP2A ITAM modulated rather than inhibited BCR-induced Ca²⁺ mobilization.

Our data establish that LMP2A expression has a function beyond the putative inhibition of the BCR by generating a ligand-independent cellular activation signal that may provide a molecular switch for different EBV life cycle stages and most probably contributes to EBV-associated lymphoproliferative disorders.

Latent membrane protein 2A (LMP2A), expressed in most Epstein-Barr virus (EBV)-associated malignancies, has been demonstrated to be responsible for the maintenance of latent infection and epithelial cell transformation. Besides, it could also act as the target for a CTL-based therapy for EBV-associated malignancies. In the present study, sequence variations of LMP2A in EBV-associated gastric carcinoma (EBVaGC) and healthy EBV carriers from Guangzhou, southern China, where nasopharyngeal carcinoma (NPC) is endemic, were investigated. Widespread sequence variations in the LMP2A gene were found, with no sequence identical to the B95.8 prototype. No consistent mutation was detected in all isolates. The immunoreceptor tyrosine-based activation motif (ITAM) and PY motifs in the amino terminus of LMP2A were strictly conserved, suggesting their important roles in virus infection; while 8 of the 17 identified CTL epitopes in the transmembrane region of LMP2A were affected by at least one point mutation, which may implicate that the effect of LMP2A polymorphisms should be considered when LMP2A-targeted immunotherapy is conducted. The polymorphisms of LMP2A in EBVaGC in gastric remnant carcinoma (GRC) were for the first time investigated in the world. The LMP2A sequence variations in EBVaGC in GRC were somewhat different from those in EBVaGC in conventional gastric carcinoma. The sequence variations of LMP2A in EBVaGC were similar to those in throat washing of healthy EBV carriers, indicating that these variations are due to geographic-associated polymorphisms rather than EBVaGC-associated mutations. This, to our best knowledge, is the first detailed investigation of LMP2A polymorphisms in EBVaGC in Guangzhou, southern China, where NPC is endemic.

• Adult
• Aged
• Carcinoma/pathology
• Carcinoma/virology
• China
• Epitopes/immunology
• Exons
• Female
• Herpesvirus 4, Human/genetics
• Herpesvirus 4, Human/isolation & purification
• Herpesvirus 4, Human/metabolism
• Humans
• Male
• Middle Aged
• Polymorphism, Genetic
• Stomach Neoplasms/pathology
• Stomach Neoplasms/virology
• T-Lymphocytes, Cytotoxic/immunology
• Viral Matrix Proteins/genetics
• Viral Matrix Proteins/metabolism
• Young Adult

• epstein-barr virus
• ebv
• microrna
• mirna
• b cell lymphoma
• lmp1
• mir-155
• mir-34
• mir-146
• mir-200
• lytic reactivation
• zeb

• Animals
• B-Lymphocytes/immunology
• B-Lymphocytes/metabolism
• B-Lymphocytes/virology
• Conserved Sequence
• Epstein-Barr Virus Infections/complications
• Epstein-Barr Virus Infections/immunology
• Epstein-Barr Virus Infections/virology
• Exosomes/immunology
• Exosomes/metabolism
• Exosomes/virology
• Gene Expression Profiling
• Gene Expression Regulation, Viral
• Herpesvirus 4, Human/genetics
• Herpesvirus 4, Human/immunology
• Herpesvirus 4, Human/metabolism
• Host-Pathogen Interactions
• Humans
• Lymphoma, B-Cell/etiology
• Lymphoma, B-Cell/immunology
• Lymphoma, B-Cell/virology
• Mice
• MicroRNAs/genetics
• MicroRNAs/immunology
• MicroRNAs/metabolism
• Oligonucleotide Array Sequence Analysis
• RNA, Messenger/genetics
• RNA, Messenger/immunology
• RNA, Messenger/metabolism
• RNA, Viral/genetics
• RNA, Viral/immunology
• RNA, Viral/metabolism
• RNA-Induced Silencing Complex/immunology
• Viral Proteins/genetics
• Viral Proteins/immunology
• Viral Proteins/metabolism
• Virus Activation/physiology
• Virus Latency/physiology

• P30 AI045008 NIAID NIH HHS
• R01 CA140337 NCI NIH HHS
• P30-AI045008 NIAID NIH HHS

To evaluate the possible effects of LMP2A (EBV latent membrane protein 2A) on human gastric cancer cell line SGC-7901, LMP2A coding gene was subcloned into shuttle plasmid pAdTrackCMV to form transfer plasmid pAdTrackCMV-2A, which was linearized with PmeI and cotransformed into E.coli BJ5183 with adenovirus genomic plasmid of pAdeasy-1. The identified recombinant adenovirus plasmid DNA was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles named vAd-2A. Then the expression and antiapoptosis activities of LMP2A on SGC-7901 infected with vAd-2A were analyzed. The vAd-2A was successfully constructed and identified by PCR, restriction digestion, and sequencing. LMP2A expression in SGC was identified by strong green fluorescence expression with fluorescence microscopic photograph and Southern blotting. The growth of LMP2A expressing SGC cells was apparently improved. Both cyclin E expression and S phase ratio in LMP2A expressing SGC cells were upregulated by cell cycle analysis and confocal microscopic analysis respectively. The replication-deficient recombinant adenovirus vector can express LMP2A antigen in SGC cells and inhibit their apoptosis. The results indicate that LMP2A might play an important role in pathogenesis of EBV-associated gastric cancer (EBVaGC). This study establishes a foundation for further study on EBVaGC and its gene therapy.

• Cell Transformation, Neoplastic/pathology
• Epstein-Barr Virus Infections/complications
• Epstein-Barr Virus Infections/pathology
• Humans
• Lymphoma, Extranodal NK-T-Cell/pathology
• Lymphoma, Extranodal NK-T-Cell/virology
• Lymphoma, T-Cell, Peripheral/pathology
• Lymphoma, T-Cell, Peripheral/virology

• Animals
• Herpesvirus 4, Human/physiology
• Herpesvirus 8, Human/physiology
• Human T-lymphotropic virus 1/physiology
• Humans
• NF-kappa B/physiology
• Oncogene Proteins, Viral/physiology
• Signal Transduction

• GM084459 NIGMS NIH HHS
• R01 CA068939 NCI NIH HHS
• AI064639 NIAID NIH HHS
• R29 CA068939 NCI NIH HHS
• CA068939 NCI NIH HHS
• R37 AI064639 NIAID NIH HHS
• R01 AI064639 NIAID NIH HHS
• R01 CA103646 NCI NIH HHS
• AI057555 NIAID NIH HHS
• R01 GM084459 NIGMS NIH HHS
• R01 AI057555 NIAID NIH HHS
• CA103646 NCI NIH HHS

• gamma-herpesviruses
• vaccine
• ebv
• kshv
• mhv-68

• Animals
• Cell Transformation, Neoplastic/genetics
• Cell Transformation, Viral/genetics
• Disease Models, Animal
• Gammaherpesvirinae/pathogenicity
• Gammaherpesvirinae/physiology
• Herpesviridae Infections/immunology
• Herpesviridae Infections/prevention & control
• Herpesviridae Infections/virology
• Herpesvirus Vaccines/genetics
• Humans
• Immune Evasion
• Mice
• Tumor Virus Infections/immunology
• Tumor Virus Infections/prevention & control
• Tumor Virus Infections/virology
• Vaccination/trends
• Vaccines, Attenuated/genetics
• Virus Latency/genetics
• Virus Latency/immunology
• Virus Replication

• R01 DE015752 NIDCR NIH HHS
• DE18337 NIDCR NIH HHS
• DE15752 NIDCR NIH HHS
• F32AI084327 NIAID NIH HHS
• P01 DE019085 NIDCR NIH HHS
• RC2 CA148250 NCI NIH HHS
• AI42927 NIAID NIH HHS
• R21 DE018337 NIDCR NIH HHS
• CA148250 NCI NIH HHS
• DE19085 NIDCR NIH HHS
• T32 AI49823 NIAID NIH HHS
• R01 AI042927 NIAID NIH HHS