Analysis Of Epstein-Barr Virus Research Articles

Establishing workflow for locoregionally advanced nasopharyngeal carcinoma (NPC) patients undergoing induction chemotherapy (IC) prior to radiation therapy (RT)

Purpose: Treatment management for locoregionally advanced nasopharyngeal carcinoma (NPC) patients involves concurrent radiation therapy (RT) and chemotherapy. Phase III clinical trial by Sun Yat-sen University Cancer Center, introduced Gemcitabine (Gem) and Cisplatin (Cis) as induction chemotherapy (IC) regime prior to concurrent chemo-radiotherapy (CCRT). This study reported 3-year recurrence-free survival of 85.3% in IC followed by CCRT, which corresponded to 4.3% higher overall survival at 3 years among IC group compared to standard therapy group. In July 2019, IC GemCis followed by CCRT was adopted as standard of care for locoregionally advanced NPC patients in National Cancer Centre Singapore (NCCS), Department of Radiation Oncology (DRO). To enable prompt referral of patients back to DRO for CCRT, new workflow was established.Methods: Since July 2019, >120 NPC patients were included in the pathway of IC and CCRT. Their journey through IC cycles were tracked as part of the newly established workflow and DRO bookings were made between second and third cycles of IC regime. Besides, their original shells were kept to replicate the original position and head tilt of pre- and post- IC.Results: IC NPC patients experienced a prompt referral back to DRO due to precise tracking of patients’ IC journey. The established workflow was found to be robust and beneficial in preventing prolonged delay to facilitate the timely commencement of CCRT. The original shell that helped to replicate the original position allows optimized delineation of gross tumor volume (GTV) post-IC, reduce workload for re-making shell and saves money plus resources.Conclusion: With the success of this robust workflow, valuable database could be obtained to enable future studies. Moving forward, analysis of Epstein-Barr virus (EBV) load, pre- and post-IC GTV, along with evaluation of dosimetric impact on surrounding organs at risk (OARs) will be investigated. Purpose: Treatment management for locoregionally advanced nasopharyngeal carcinoma (NPC) patients involves concurrent radiation therapy (RT) and chemotherapy. Phase III clinical trial by Sun Yat-sen University Cancer Center, introduced Gemcitabine (Gem) and Cisplatin (Cis) as induction chemotherapy (IC) regime prior to concurrent chemo-radiotherapy (CCRT). This study reported 3-year recurrence-free survival of 85.3% in IC followed by CCRT, which corresponded to 4.3% higher overall survival at 3 years among IC group compared to standard therapy group. In July 2019, IC GemCis followed by CCRT was adopted as standard of care for locoregionally advanced NPC patients in National Cancer Centre Singapore (NCCS), Department of Radiation Oncology (DRO). To enable prompt referral of patients back to DRO for CCRT, new workflow was established. Methods: Since July 2019, >120 NPC patients were included in the pathway of IC and CCRT. Their journey through IC cycles were tracked as part of the newly established workflow and DRO bookings were made between second and third cycles of IC regime. Besides, their original shells were kept to replicate the original position and head tilt of pre- and post- IC. Results: IC NPC patients experienced a prompt referral back to DRO due to precise tracking of patients’ IC journey. The established workflow was found to be robust and beneficial in preventing prolonged delay to facilitate the timely commencement of CCRT. The original shell that helped to replicate the original position allows optimized delineation of gross tumor volume (GTV) post-IC, reduce workload for re-making shell and saves money plus resources. Conclusion: With the success of this robust workflow, valuable database could be obtained to enable future studies. Moving forward, analysis of Epstein-Barr virus (EBV) load, pre- and post-IC GTV, along with evaluation of dosimetric impact on surrounding organs at risk (OARs) will be investigated.

Lessons from Epstein-Barr virus DNA detection in cerebrospinal fluid as a diagnostic tool for EBV-induced central nervous system dysfunction among HIV-positive patients.

Polymerase chain reaction (PCR) analysis of Epstein-Barr virus (EBV) DNA in cerebrospinal fluid (CSF) remains vital for evaluating active EBV infection involving the central nervous system (CNS). CSF EBV DNA was often found in conjunction with other microbial infection affecting the CNS among patients infected with human immunodeficiency virus (HIV). Sometimes CSF EBV DNA is detectable in patients without neurological symptoms. This review focused on the clinical and laboratory features of CNS EBV infection among patients with HIV, and discussed various types of EBV-associated CNS infections, and predominant neoplasms involving CNS such as primary central nervous system lymphoma (PCNSL), CNS-non-Hodgkin's lymphoma, smooth muscle tumors and leiomyosarcomas, EBV encephalitis or myelitis, EBV meningitis and EBV coinfection with other causative agents were also included. Furthermore, the metagenomic next-generation sequencing technique with high sensitivity for the detection of pathogenic coinfection in the CSF were also reviewed. We concluded that CSF EBV-DNA detection with high sensitivity and specificity could be a useful diagnostic tool for CNS lymphoma among HIV patients; however, it is still unknown for other CNS diseases. We further summarized and conclude that positive CSF EBV-DNA detection combined with specific brain focal lesions could be a minimally invasive method to diagnose PCNSL. The occurrence of positive CSF EBV-DNA was influenced by PCR detection limit, PCR methods, immunocompromised status, the possible influence of anti-herpetic therapy and anti-HIV therapy, and the size and location of a tumor mass. Uniform PCR methods as vital diagnostic tools and optimal EBV-DNA load threshold need to be established.

Epstein-Barr virus-coded miR-BART13 promotes nasopharyngeal carcinoma cell growth and metastasis via targeting of the NKIRAS2/NF-κB pathway

Based on analysis of Epstein-Barr virus (EBV) BART microRNA expression profiles, we previously reported that EBV-encoded miR-BART13 is upregulated in nasopharyngeal carcinoma (NPC) plasma specimens. However, the effects and molecular mechanisms of miR-BART13 in NPC remain largely unknown. We found that miR-BART13 was significantly upregulated in NPC tissue specimens. Ectopic expression of miR-BART13 promoted NPC cell proliferation, epithelial mesenchymal transition, and metastasis in vitro, and facilitated xenograft tumor growth and lung metastasis in vivo. Molecularly, NF-κB inhibitor interacting Ras-like 2 (NKIRAS2), a negative regulator of the NF-κB signaling, was identified to be a direct target of miR-BART13 in NPC cells, and NKIRAS2 mRNA and protein expression was inversely correlated with miR-BART13 in NPC tissues, respecitvely. Furthermore, the NF-κB signaling pathway was activated by miR-BART13. By rescued experiments, reconstitution of NKIRAS2 expression abrogated all the phenotypes upregulated by miR-BART13, and attenuated activity of NF-κB signaling pathway activated by miR-BART13 in NPC cells. Our findings indicated the newly identified miR-BART13/NKIRAS2/NF-κB signaling axis may provide further insights into better understanding of NPC initiation and development, and targeting of this pathway could be further studied as a therapeutic strategy for NPC patients.

Regulatory network analysis of Epstein-Barr virus identifies functional modules and hub genes involved in infectious mononucleosis.

Epstein-Barr virus (EBV) is the most common cause of infectious mononucleosis (IM) and establishes lifetime infection associated with a variety of cancers and autoimmune diseases. The aim of this study was to develop an integrative gene regulatory network (GRN) approach and overlying gene expression data to identify the representative subnetworks for IM and EBV latent infection (LI). After identifying differentially expressed genes (DEGs) in both IM and LI gene expression profiles, functional annotations were applied using gene ontology (GO) and BiNGO tools, and construction of GRNs, topological analysis and identification of modules were carried out using several plugins of Cytoscape. In parallel, a human-EBV GRN was generated using the Hu-Vir database for further analyses. Our analysis revealed that the majority of DEGs in both IM and LI were involved in cell-cycle and DNA repair processes. However, these genes showed a significant negative correlation in the IM and LI states. Furthermore, cyclin-dependent kinase 2 (CDK2) - a hub gene with the highest centralityscore - appeared to be the key player in cell cycle regulation in IM disease. The most significant functional modules in the IM and LI states were involved in the regulation of the cell cycle and apoptosis, respectively. Human-EBV network analysis revealed several direct targets of EBV proteins during IM disease. Our study provides an important first report on the response to IM/LI EBV infection in humans. An important aspect of our data was the upregulation of genes associated with cell cycle progression and proliferation.

Analysis of Epstein-Barr virus and cellular gene expression during the early phases of Epstein-Barr virus lytic induction.

In order to develop novel host/pathogen real-time PCR assays for routine diagnostic use, early gene expression patterns from both Epstein-Barr virus (EBV) and Raji cells were examined after inducing the lytic life cycle using 12-O-tetradecanoyl-13-phorbol ester and sodium butyrate. Real-time PCR identified several highly induced (>90-fold) EBV lytic genes over a 48 h time course during the lytic induction phase. Latent genes were induced at low levels during this phase. The cellular response to lytic viral replication is poorly understood. Whole human genome microarray analysis identified 113 cellular genes regulated twofold or more by EBV, including 63 upregulated and 46 downregulated genes, over a 24 h time course post-induction. The most upregulated gene was CHI3L1, a chitinase-3-like 1 protein (18.1-fold; P<0.0084), and the most downregulated gene was TYMS, a thymidylate synthetase (-7.6-fold). Gene Ontology enrichment analysis using MetaCore software revealed cell cycle (core), cell cycle (role of anaphase-promoting complex) in cell cycle regulation) and lymphatic diseases as the most significantly represented biological network processes, canonical pathways and disease biomarkers, respectively. Chemotaxis, DNA damage and inflammation (IL-4 signalling) together with lymphoproliferative disorders and non-Hodgkin's lymphoma were significantly represented biological processes and disease biomarkers.

Epstein-Barr Virus-Positive Diffuse Large B-Cell Lymphoma in the Elderly: A Matched Case-Control Analysis.

BackgroundEpstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) in the elderly has rarely been reported. This study aimed to explore the clinical characteristics and prognosis of this entity.MethodsIn situ hybridization (ISH) analysis of Epstein-Barr virus (EBV) and immunohistochemistry was performed in 230 tumor specimens from consecutive de novo DLBCL patients over 50 years old. A matched-case control analysis (1:3) was utilized to compare EBV-positive and EBV-negative DLBCL in the elderly.ResultsA total of 16 patients (7.0%) were diagnosed with EBV-positive DLBCL. Of these 16 cases, the median age was 62 years, with a male to female ratio of 11:5. Elderly EBV-positive DLBCL patients had a higher incidence of non-germinal center B-cell (non-GCB) subtypes (87.5%) and high Ki67 (75%) and CD30 expression (93.8%). For EBV-positive patients undergoing initial chemotherapy, 7 of 16 (43.8%) had complete remission, 2 (12.5%) had partial remission, 2 (12.5%) had stable disease, and 5 (31.3%) had progressive disease. The median overall survival was 9 months for the EBV-positive patients. A matched-case control analysis suggested that EBV-positive patients had inferior survival outcomes compared with EBV-negative patients (3-year progression-free survival [PFS]: 25% vs. 76.7%, respectively; 3-year overall survival [OS]: 25% vs. 77.4%, respectively; P<0.001).ConclusionEBV-positive DLBCL of the elderly is associated with an inferior clinical course and inferior survival outcomes. The role of EBV in this disease and the optimal management of this subgroup warrants further investigation.

The effect of anti-CD40 ligand antibody on B cells in human systemic lupus erythematosus.

To determine the immunologic effects of anti-CD40 ligand (anti-CD40L) therapy in 5 patients with systemic lupus erythematosus nephritis who participated in an open-label study of a humanized anti-CD40L monoclonal antibody. Serum and peripheral blood mononuclear cells were obtained before, during, and after treatment, and the frequency of Ig and anti-DNA antibody-secreting B cells was analyzed by enzyme-linked immunospot assay and by analysis of Epstein-Barr virus (EBV)-transformed B cell lines. To determine the effect of treatment on somatic mutation of Ig genes, reverse transcriptase-polymerase chain reaction was performed on messenger RNA from 4 patients, using primers specific for the DP-47 heavy chain gene and for IgG. Finally, B cell phenotype was investigated using flow cytometry. Even a brief period of treatment with anti-CD40L markedly reduced the frequency of IgG and IgG anti-DNA antibody-producing B cells, and these changes persisted for several months after cessation of treatment. To confirm these findings, EBV-transformed B cell lines were screened from each of 3 patients, and a 10-fold decrease in anti-DNA antibody-secreting cell lines was found after treatment in all 3 patients. Few differences in mutation patterns were observed before and after treatment; however, the frequency of germline-encoded DP-47 sequences was significantly increased before treatment and normalized following treatment. Flow cytometric analysis of B cells revealed expansion of a CD27-/IgD- B cell subset in some of the patients, which did not change with treatment. These are the first mechanistic studies of the effect of anti-CD40L therapy in human autoimmune disease. The results suggest that further studies of CD40L blockade are warranted.

Analysis of Epstein-Barr virus (EBV) nuclear antigen 1 subtypes in EBV-associated lymphomas from Brazil and the United Kingdom.

EBNA-1 is the only viral protein consistently expressed in all cells latently infected by Epstein-Barr virus (EBV). There is a high frequency of sequence variation within functionally important domains of EBNA-1, with five subtypes identified. Individuals may be infected with multiple EBV strains (classified according to EBNA-1 subtype), but Burkitt's lymphoma (BL) tumours carry a single subtype and exhibit some subtype preference. Subtype variation has also been related to geographical location. In the present study EBNA-1 polymorphisms were examined in a series of haematological malignancies from two distinct geographical regions, Brazil and the United Kingdom. Nucleotide sequence analysis of the carboxy-terminal region of EBNA-1 in 34 cases revealed six distinct sequences, some of which are novel. A new subtype, named V-Ala, was identified. EBNA-1 subtype in tumours differed markedly according to geographical location. In contrast to previous studies, we found evidence of EBNA-1 sequence variation within individual BL tumour samples.

ホジキン病での各種ＥＢウイルス検出法の検討

ホジキン病での各種ＥＢウイルス検出法の検討

Analysis of Epstein-Barr virus in Hodgkin's disease: experience of a single university hospital in Korea.

Hodgkin's disease is known to be associated with Epstein-Barr virus (EBV) infection in Western countries, and viral nucleic acids and proteins have been identified within Reed-Sternberg (RS) cells, which are the histopathologic hallmark of the disease process. Twenty-five cases of Hodgkin's disease from a single university hospital in Korea were studied for evidence of EBV by in situ hybridization for EBV DNA and RNA and immunohistochemistry for an EBV latent protein. EBV nucleic acids were studied by a rapid (60 minutes) in situ hybridization procedure, which utilized biotinylated DNA probes specific for the following nucleic acid sequences: (1) EBV EBER1 RNA (an abundant RNA sequence expressed during latent EBV infection), (2) EBV NotI repeats (a tandemly repeated DNA sequence, which has been established to identify amplified EBV genome in lytic EBV infection), and (3) BAM HI W (a DNA sequence reiterated 11 times within the viral genome). In addition, immunohistochemistry for EBV latent membrane protein, a protein that is capable of inducing cellular transformation in cell culture, was also performed. EBV was identified within the neoplastic RS cells by at least one method in 19/25 cases (76%). The mixed cellularity subtype was the most common subtype associated with EBV infection (11/13-85%). In situ hybridization for EBV EBER1 RNA was the most sensitive method for EBV detection and was present in 17/25 cases. A significant proportion of Korean Hodgkin's disease cases is associated with EBV infection.

Analysis of Epstein-Barr virus in localized nasopharyngeal carcinoma tumors.

An f variant of Epstein-Barr virus (EBV) appears associated with nasopharyngeal carcinoma (NPC) in Southern Chinese. Early diagnosis of the tumor allows the detection of some localized tumors. A polymerase chain reaction (PCR) assay for genotyping EBV was used to evaluate the presence of the virus in NPC biopsies of local tumors of eight Chinese patients. The f variant was detected in the nasopharynx of seven of eight patients. The f variant was present in equal frequency in the "normal" and tumor regions. Examination of localized NPC tumors by the PCR genotyping assay revealed EBV was present on the tumor side of the nasopharynx in greater quantities than the "normal" side in seven of eight patients studied. Concurrent infection with both the prototype F and f variant was observed in two of the eight patients investigated.

Analysis of Epstein-Barr Virus (EBV) of P3HR-1: Isolation of EBV with EBNA induction ability in human cord lymphocytes and without EA induction ability in raji cells

A subline of P3HR-1 cells was isolated through a prolonged (over 1 year) propagation of the cells at a non-EBV-productive condition followed by cell cloning procedures. Cloned cells thus obtained, designated DHR1, produced EBV when brought back to the EBV-productive condition. Restriction enzyme analysis of the viral DNA revealed that DHR1 EBV is composed of an apparently homogeneous EBV population, and it displays a similar but not identical genome organization compared with HR-1 EBV. The characteristic biological properties of DHR1 EBV included the ability to induce EBV-associated nuclear antigen (EBNA) in human cord lymphocytes and the inability to induce EBV-associated early antigen (EA) in Raji cells. These are in striking contrast to the behavior of the parental HR-1 EBV. Thus, P3HR-1 cultures after a period of nonproductivity reinitiated the production of virus with an apparently homogeneous and unique population of EBV distinguishable from the original HR-1 virus.