Epstein-Barr Virus DNA Load Research Articles

Epstein-Barr virus DNA loads in the peripheral blood cells predict the survival of locoregionally-advanced nasopharyngeal carcinoma patients.

Objective:Circulating cell-free Epstein-Barr virus (EBV) DNA has been shown to be a valuable biomarker for population screening and prognostic surveillance for nasopharyngeal carcinoma (NPC). Despite important insights into the biology of persistence, few studies have addressed the clinical significance of cell-based EBV-DNA loads in peripheral blood cells (PBCs).Methods:A prospective observational cohort study was conducted involving 1,063 newly diagnosed, locoregionally-advanced NPC patients at Sun Yat-sen University Cancer Center from 2005 to 2007. Cox regression analysis was conducted to identify the association of PBC EBV DNA loads to overall survival (OS) and other prognostic outcomes. Prognostic nomograms were developed based on PBC EBV DNA loads to predict survival outcomes for NPC patients.Results:After a median follow-up of 108 months, patients with higher PBC EBV-DNA loads had significantly worse OS [hazard ratio (HR) of medium, medium-high, and high vs. low were 1.50, 1.52, and 1.85 respectively; Ptrend < 0.001]. Similar results were found for progression-free survival and distant metastasis-free survival. The concordance index of the prognostic nomogram for predicting OS in the training set and validation set were 0.70 and 0.66, respectively. Our data showed that the PBC EBV DNA load was an independent and robust survival biomarker, which remained significant even after adjusting for plasma EBV DNA loads in a subset of 205 patients of the cohort (HR: 1.88; P = 0.025). Importantly, a combination of PBC EBV DNA load and plasma EBV DNA load improved the predicted OS.Conclusions:The EBV-DNA load in PBCs may be an independent prognosis marker for NPC patients.

Rapid nasopharyngeal brush-smear cytology with Epstein-Barr virus DNA load and viral marker analysis for detection of nasopharyngeal carcinoma

Background: Nasopharyngeal carcinoma (NPC) is a prevalent cancer in Indonesia, usually identified at late stage. Epstein-Barr virus (EBV) gene products are present in all tumor cells. Early-stage NPC diagnosis is clinically important but difficult due to non- specific symptoms. In patients suspected of having NPC, nasopharyngeal (NP) brush-cytology may provide a simple non-invasive procedure for rapid on-site detection of NPC tumour cells that can be combined with EBV markers. Methods: Patients (N=83) suspected of having NPC underwent nasoendoscopy-guided NP brushing before a biopsy from clinically suspicious areas. The loaded brush was first smeared onto glass slides that were directly immersed in ethanol, followed by Papanicolaou (PAP) staining and on-site interpretation of tumour cell presence. Thereafter, slides were used for EBER RNA in situ hybridization (EBER-ISH) and LMP-1 immunocytochemistry (ICC). The brush left-over was used for EBV-DNA load measurement using real-time PCR. Biopsy specimens were taken from the same NP site and processed for routine pathology and EBER-ISH on tissue sections. Result: Morphological detection of NPC tumour cells on PAP-stained brush-smears gave a sensitivity and specificity of 87% and 100%, respectively. EBER-RISH and LMP1 ICC combined showed 100% positivity for NPC cases. Expression of LMP-1 was detected in 77.8% of cases. Median EBV-DNA load in left-over brush material from NPC cases was 5.06 × 106 copies/brush (range 777 to 242,400,000 copies/brush), whereas median value in non-NPC cases was 4.4 x 103 copies/brush (range 484 - 297,200; P < 0.001). Conclusion: Minimal invasive NP-brush cytology with conventional PAP staining allows rapid, simple and cheap “on site” detection of NPC tumour cells in suspected NP lesions. The method can be modified for direct demonstration of EBV involvement using EBER-ISH and LMP1-ICC or EBV-DNA load measurement. This NP brush procedure may be useful for primary screening in patients considered at risk for NPC.

Long‐term monitoring of dynamic changes in plasma EBV DNA for improved prognosis prediction of nasopharyngeal carcinoma

BackgroundThis study was performed to investigate whether long‐term monitoring of dynamic changes in plasma Epstein‐Barr virus (EBV) DNA could improve prognosis prediction of nasopharyngeal carcinoma (NPC).Materials and methodsAbout 1077 nonmetastatic NPC patients were recruited to retrospectively analyze the prognostic value of plasma EBV DNA load pretreatment and 3, 12, 24, and 36 months posttreatment. We also examined the prognostic value of dynamic changes in plasma EBV DNA at various time points.ResultsPatients with plasma EBV DNA load above optimal pre‐ and posttreatment cut‐offs had significantly worse five‐year progression‐free survival, distant metastasis‐free survival, locoregional relapse‐free survival, and overall survival (OS) at all‐time points, excluding only OS at 36 months posttreatment due to limited mortalities. Patients with persistently undetectable plasma EBV DNA at the first four time points had the best prognosis, followed by those with positive detection pretreatment and consistently negative detection posttreatment, those with negative detection pretreatment and positive detection at one time point posttreatment, and those with positive detection pretreatment and at one time point posttreatment, whereas patients with positive detection at ≥2 time points posttreatment had the worst prognosis. Cox proportional hazard models identified the dynamic change pattern as an independent prognostic factor, and receiver operating characteristic curve analysis demonstrated that the dynamic change at four time point was more valuable than any single time point for predicting disease progression, distant metastasis, locoregional relapse, and mortality.ConclusionsDynamic changes in plasma EBV DNA pre‐ and posttreatment could predict the long‐term survival outcome and provide accurate risk stratification in NPC.

Effect of PD-L1 and tumor regression rate on prognosis of nasopharyngeal carcinoma.

To explore the expression level of programmed death ligand-1 (PD-L1) and the tumor regression rate after chemoradiotherapy, and to investigate their influence on the prognosis of patients with nasopharyngeal carcinoma. Eighty-six patients, who were initially diagnosed with non-metastatic nasopharyngeal carcinoma in the Second Xiangya Hospital of Central South University from January 2014 to June 2014, were recruited. Positive expression of PD-L1 in prepared sections of nasopharyngeal carcinoma tissues was detected by immunohistochemical staining. The clinical data of recruited patients and the tumor regression rate were calculated before chemoradiotherapy and 1 month after chemoradiotherapy. In addition, the potential influence of PD-L1 level and the tumor regression rate after chemoradiotherapy on therapeutic efficacy and the prognosis of nasopharyngeal carcinoma were assessed. Among the 86 patients with nasopharyngeal carcinoma, there were 30 patients with high expression of PD-L1 in nasopharyngeal carcinoma sections (34.88%), while there were 56 patients with low expression (65.12%). Expression levels of PD-L1 in nasopharyngeal carcinoma tissues were not related with sex, age, smoking history, concurrent chemoradiotherapy, pathological type, N staging, tumor staging, and pre-treatment EBV-DNA load in peripheral blood in the affected patients (all P>0.05). But the expression level of PD-L1 was related to the tumor regression rate and T staging in the patients with nasopharyngeal carcinoma (both P<0.05). The expression levels of PD-L1 in nasopharyngeal carcinoma tissues was a risk factor for tumor regression rate (OR=0.226, P<0.05). Compared with the patients with low expression levels of PD-L1, there was shorter disease-free survival in the patients with high expression levels of PD-L1 (P<0.05). Moreover, compared with the patients with low tumor regression rate, the nasopharyngeal carcinoma patients with high tumor regression rate showed a favorable disease-free survival (P<0.05). A high expression level of PD-L1 and a low tumor regression rate after chemoradiotherapy are unfavorable to the prognosis of nasopharyngeal carcinoma.

Clinical application of Epstein-Barr virus DNA loads in Epstein-Barr virus-associated diseases: A cohort study

ObjectivesThis study evaluated the diagnostic value of Epstein-Barr virus (EBV) DNA load in blood samples of patients with EBV-associated diseases, and proposed a strategy for the interpretation of positive EBV DNA results. MethodsDerivation and validation cohorts were established to evaluate the clinical significance of EBV DNA loads in the peripheral blood mononuclear cells (PBMCs) and plasma from EBV-infected patients. EBV DNA loads were compared and receiver operating characteristic curves were employed to assess the optimal cutoff values of EBV DNA for identification of EBV-associated diseases. ResultsThe derivation and validation cohorts comprised 135 and 71 subjects, respectively. EBV DNA loads in the PBMCs of the EBV-associated diseases group was significantly higher than that of the EBV non-associated diseases group (5.8 × 104 vs 7.8 × 103 copies/106 cells, P<0.0001). The diagnostic cut-off value of viral load in PBMCs for EBV-associated diseases was determined to be 1.6 × 104 copies/106 cells. The combined EBV DNA load cutoff in PBMCs and positive EBV DNA qualitative detection in plasma (>500 copies/mL) allowed for the differentiation of EBV-associated and non-associated diseases; the sensitivity and specificity were 80.6 and 96.8%, respectively. ConclusionsThe strategy of combining EBV DNA loads in PBMCs and plasma will potentially help identify EBV-associated diseases.

The association of Epstein-Barr virus infection with CXCR3+ B-cell development in multiple sclerosis: impact of immunotherapies.

Epstein–Barr virus (EBV) infection of B cells is associated with increased multiple sclerosis (MS) susceptibility. Recently, we found that CXCR3‐expressing B cells preferentially infiltrate the CNS of MS patients. In chronic virus‐infected mice, these types of B cells are sustained and show increased antiviral responsiveness. How EBV persistence in B cells influences their development remains unclear. First, we analyzed ex vivo B‐cell subsets from MS patients who received autologous bone marrow transplantation (n = 9), which is often accompanied by EBV reactivation. The frequencies of nonclass‐switched and class‐switched memory B cells were reduced at 3–7 months, while only class‐switched B cells returned back to baseline at 24–36 months posttransplantation. At these time points, EBV DNA load positively correlated to the frequency of CXCR3+, and not CXCR4+ or CXCR5+, class‐switched B cells. Second, for CXCR3+ memory B cells trapped within the blood of MS patients treated with natalizumab (anti‐VLA‐4 antibody n = 15), latent EBV infection corresponded to enhanced in vitro formation of anti‐EBNA1 IgG‐secreting plasma cells under GC‐like conditions. These findings imply that EBV persistence in B cells potentiates brain‐homing and antibody‐producing CXCR3+ subsets in MS.

Chronic Active Epstein-Barr Virus Infection: Is It Immunodeficiency, Malignancy, or Both?

Simple SummaryChronic active Epstein–Barr virus (EBV) infection (CAEBV) is a rare syndrome of unknown etiology characterized by prolonged infectious mononucleosis-like symptoms and proliferation of EBV-infected T and/or natural killer cells. CAEBV has been primarily reported in East Asia and Latin America, suggesting a genetic predisposition in its pathogenesis. The clinical course of CAEBV is heterogeneous ranging from an indolent and occasionally self-limiting disease to an aggressive and fatal condition, but its prognosis is generally poor. This heterogeneous clinical picture does not suggest a simple etiology for the syndrome. Clinicopathological investigations of CAEBV suggest that it has aspects of both malignant neoplasm and immunodeficiency. This article summarizes the latest findings on CAEBV and discusses critical unsolved questions regarding its pathogenesis and disease concept.Chronic active Epstein–Barr virus (EBV) infection (CAEBV) is a rare syndrome characterized by prolonged infectious mononucleosis-like symptoms and elevated peripheral blood EBV DNA load in apparently immunocompetent persons. CAEBV has been primarily reported in East Asia and Latin America, suggesting a genetic predisposition in its pathogenesis. In most cases of CAEBV, EBV induces proliferation of its unusual host cells, T or natural killer (NK) cells. The clinical course of CAEBV is heterogeneous; some patients show an indolent course, remaining in a stable condition for years, whereas others show an aggressive course with a fatal outcome due to hemophagocytic lymphohistiocytosis, multiple organ failure, or progression to leukemia/lymphoma. The pathogenesis of CAEBV is unclear and clinicopathological investigations suggest that it has aspects of both malignant neoplasm and immunodeficiency. Recent genetic analyses of both viral and host genomes in CAEBV patients have led to discoveries that are improving our understanding of the nature of this syndrome. This article summarizes the latest findings on CAEBV and discusses critical unsolved questions regarding its pathogenesis and disease concept.

The Impact of Epstein-Barr Virus Infection on Juvenile Idiopathic Arthritis Activity and Patient's Response to Treatment.

This study aimed to investigate the relationship between Epstein-Barr virus (EBV) infection and the onset of juvenile idiopathic arthritis (JIA), disease activity, and response to treatment. The study included 44 children with JIA, 23 children with different types of arthritis, and 44 controls. We measured EBV infection markers, including the EBV DNA load and the concentration of antibodies to viral antigens, at disease onset, before treatment. Six months after JIA diagnosis and the initiation of treatment patients with anti-viral capsid antigen IgG had a higher disease activity and worse response to treatment than patients without previous infection. After six months of treatment, the probability of disease inactivity in children without a history of EBV infection was almost 6.5 times greater than in a child with a history of infection. Furthermore, the probability of a better response after six months of treatment in a child with a history of EBV infection was more than five times smaller than in a child without infection. A past EBV infection can have a negative effect on achieving disease remission and may be associated with a worse response to treatment. Our results do not indicate the need for routine assessment of EBV infection markers in patients with JIA.

Clinical Characteristics of Hemophagocytic Lymphohistiocytosis Associated with Non-Hodgkin B-Cell Lymphoma: A Multicenter Retrospective Study

BackgroundHemophagocytic lymphohistiocytosis (HLH) associated with B-cell lymphoma is a highly aggressive disease with unclear clinical features and has no standard treatment. Patients and MethodsWe analyzed the clinical characteristics of 31 patients from two individual centers. ResultsThe median overall survival was only 1.5 months. Both univariate and multivariate analyses, based on lymphoma or HLH-related characteristics, revealed that patients with high Epstein-Barr virus (EBV) DNA load and ≥ 2 extranodal lesions, or hypofibrinogenemia, respectively, showed significantly poorer overall survival. Interestingly, some patients with high EBV DNA load had EBV-positive natural killer (NK) and/or T cells, which may be related to the coexistence of immunodeficiency and/or chronic active EBV infection. Molecular genetics examination confirmed that 47.4% (9/19) of patients had complex karyotypes, 37.5% (3/8) of patients had TP53 deletions, and 21.34% (3/14) of patients had TP53 mutation or alteration of malignancy-related pathways, including BCR/NF-κB, JAK-STAT, and epigenetic regulatory pathways, which may provide clues to choose targets for therapy. Treatment regimens containing etoposide, anti-CD20 monoclonal antibodies, or anthracyclines improved patient prognosis (P = .0183, .025, and .0436, respectively). Patients with infections had significantly shorter survival than those without infections (P = .00019). ConclusionThe patients’ performance status, number of extranodal lesions, high EBV DNA load, and hypofibrinogenemia are poor prognostic factors for HLH associated with B-cell lymphoma. Molecular genetic high-risk factors are of particular importance because these factors can provide information for prognosis prediction, treatment decisions, and disease surveillance.

Expression of pSTAT3 and PD-L1 in extranodal NK/T cell lymphoma and its clinical significance

Objective: To study the expression of phosphates signal transducer and activator of transcription 3 (pSTAT3) and programmed death ligand-1 (PD-L1) in extranodal NK/T cell lymphomas (ENKTCL) and the relationships of pSTAT3 and PD-L1 expression with the clinicopathological characteristics and prognosis of ENKTCL. Methods: Fifty-one cases of ENKTCL diagnosed at Guangdong Provincial People's Hospital from June 2015 to February 2019 were included in the study. The expression of pSTAT3 and PD-L1 was examined using immunohistochemistry. Results: There were 35 males and 16 females, ranging from 18 to 85 years old with a median age of 47 years. The positive rates of pSTAT3 and PD-L1 expression were 68.6% (35/51) and 76.5% (39/51), respectively. pSTAT3 expression was correlated with PD-L1 expression (P=0.033,R=0.322), while there were no associations of pSTAT3 and PD-L1 expression with the clinicopathological characteristics of ENKTCL, including age, sex, clinical site, B symptom, Ann Arbor stage, LDH value, EBV DNA load of peripheral blood and international proliferation index score. Kaplan-Meier survival analysis showed the prognoses of the pSTAT3 and PD-L1 positive groups were slightly better than the respective negative groups, but the differences were not significantly (P>0.05). Conclusions: pSTAT3 is highly expressed in extranodal NK/T cell lymphoma and related to the expression of PD-L1, which provides a potential target and rationale for combinations of targeted therapies and immune checkpoint blockade inhibitors in the treatment of ENKTCL.

Abstract CT203: A phase II trial of ipilimumab in combination with nivolumab in EBV-associated advanced nasopharyngeal carcinoma (NCT03097939)

Abstract Background: Single agent PD-1/PD-L1 studies in previously treated EBV-associated nasopharyngeal carcinoma (NPC) demonstrate clinical outcomes inferior to salvage combination chemotherapy. Dual blockade of PD-1/CTLA4 is a viable treatment strategy in other solid tumors. We hypothesized that this strategy would also be feasible and efficacious in NPC. Materials and Methods: A single arm phase 2 study using a Simon 2-stage design was used. Nivolumab was dosed at 3 mg/kg q2 weeks, and Ipilimumab was dosed at 1 mg/kg q6 weeks. Eligible pts had EBER-ISH positive NPC, measurable plasma EBV DNA, no more than 1 prior line of chemotherapy, ECOG 0-1, and adequate organ function. All pts who met the eligibility criteria and received at least one dose of the combination were included in the safety and efficacy analysis. The primary efficacy endpoint was best overall response (BOR) by RECIST 1.1. Toxicity was assessed using CTCAE criteria. Pretreatment EBV DNA load was used to discriminate EBVhi from EBVlo at a threshold of 30,000 copies/ml, and sub-group analyses were carried out for BOR, time to progression, progression-free survival and overall survival based on this cutoff. Paired tumor and blood sampling were done at baseline and on-treatment and results are presented separately. Results: A total of 28 patients were enrolled and 26 were evaluable. Two patients were excluded from analysis for eligibility reason and consent withdrawal. Median age of pts was 56 years (range 23-73). Most patients (85%) were of Chinese ethnicity and 19 patients (73%) were male. The median number of cycles received was 4. Three patients remain on treatment. Twenty-one patients (81%) experienced any grade treatment-related adverse events (trAE). Common trAEs were maculopapular rash (n=8; 31%) and hypothyroidism (n=8; 31%). Three pts (11%) required treatment discontinuation due to grade 3/4 AE, including pneumonitis and myasthenia gravis. In stage one, of 15 pts recruited, 7 reported BOR of PR (47%) and another 11 patients were recruited into stage two. In total, 8 out of 26 patients achieved PR (BOR 31%; 95% CI 14.3% to 51.8%). Median duration of response (DOR) was 5.9 mths (95% CI 3.9 to 9.0). With a median follow up of 10.6 mths, median PFS was 5.3 mths (95% CI 2.8 to 6.4). Of EBVlo pts, 8 experienced a PR (53%; 95% CI 26.6% to 78.7%). No responses were observed in EBVhi pts. EBVlo pts had a median PFS of 6.8 mths (95% CI 2.8 to 10.4) compared to EBVhi 2.7 mths (95% CI 1.7 to 5.2). Conclusions: Dual PD-1/CTLA4 blockade is safe and feasible in NPC, achieving durable responses in pts with lower plasma EBV DNA. Efficacy was comparable to that seen in other solid tumors using this combination. The trial has been expanded to further study efficacy of this combination in NPC. Citation Format: Darren Wan-Teck Lim, Quan Sing Ng, Ruey-Long Hong, Daniel S. Tan, Eng-Huat Tan, Boon-Cher Goh, Wan Ling Tan, Stella Li-Li Chan, Sze-Huey Tan, Hsiang-Fong Kao, Gopalakishna N. Iyer, Mei-Kim Ang. A phase II trial of ipilimumab in combination with nivolumab in EBV-associated advanced nasopharyngeal carcinoma (NCT03097939) [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr CT203.

TLR4 896A/G and TLR9 1174G/A polymorphisms are associated with the risk of infectious mononucleosis

Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns and activate innate and adaptive immune responses. Single nucleotide polymorphisms (SNPs) within the TLR genes may influence host–pathogen interactions and can have an impact on the progression of infectious diseases. The present study aimed to investigate the genotype distribution of TLR2 (2029C/T, rs121917864; 2258G/A, rs5743708), TLR4 (896A/G, rs4986790), and TLR9 (− 1237T/C, rs5743836; − 1486T/C, rs187084; 1174G/A, rs352139; and 2848C/T, rs352140) polymorphisms in 149 children and adolescents with infectious mononucleosis (IM) and 140 healthy individuals. The potential association of TLR SNPs with the clinical manifestations of EBV infection was also studied. The presence of TLR2, TLR4, and TLR9 SNPs was identified by polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP). EBV DNA loads were detected by quantitative real-time PCR assay. The TLR4 896 GG and the TLR9 1174 GA genotypes were associated with an increased risk of EBV-related IM in examined patients (p = 0.014 and p = 0.001, respectively). The heterozygous genotype of the TLR4 896A/G SNP was associated with an increased risk of elevated liver enzyme levels and leukocytosis (p < 0.05). Our preliminary study revealed that the TLR4 896A/G and the TLR9 1174G/A polymorphisms seem to be related to the course of acute EBV infection in children and adolescents.

Nasopharyngeal Epstein-Barr virus DNA loads in high-risk nasopharyngeal carcinoma families: Familial aggregation and host heritability.

Nasopharyngeal carcinoma (NPC), the most common head and neck cancer, is characterized by distinct geographic distribution and familial aggregation. Multiple risk factors, including host genetics, environmental factor, and EBV infection, have been linked to the development of NPC, particularly in the familial clustering cases. However, the cause of NPC endemicity remains enigmatic due possibly to the complicated interplay between these risk factors. Recently, positive Epstein‐Barr virus (EBV) DNA loads at nasopharyngeal (NP) cavity has been found to reflect NPC development and applied in NPC screening. To examine whether the increased NP EBV loads could aggregate in the families and be affected by host genetics and environmental factor, EBV loads were obtained by 510 NP brushing samples from eligible unaffected individuals, who have two or more relatives affected with NPC, in 116 high‐risk NPC families. The correlation of relative pairs was estimated using S.A.G.E. (version 6.4, 2016), and host heritability of NP EBV loads was calculated with variance component models using SOLAR (version 8.4.2, 2019). In result, significant correlations of EBV loads were observed between parent‐offspring pairs and sibling‐sibling pairs (P < .001), but not in distant kin relationship pairs. Interestingly, after excluding the shared environmental factor within families, host genetics contributes significantly to NP EBV loads with a heritability of 56.41% (P = 1.00 × 10−7), and its effect was slightly elevated (68.86%, P = 3.40 × 10−6) in families with more NPC cases (≥3). These findings indicate that additional host‐genetic variants involved in the EBV local NP mucosal behavior may be especially important for the development of NPC.

Chronic Active Epstein-Barr Virus-Associated Enteritis: CT Findings and Clinical Manifestation.

Aim To improve the identification and computed tomography (CT) diagnostic accuracy of chronic active Epstein-Barr virus (EBV)-associated enteritis (CAEAE) by evaluating its CT findings and clinical manifestation. Methods The data of three patients with pathologically and clinically confirmed CAEAE who underwent CT enterography (CTE) were retrospectively reviewed from January 2018 to October 2019. The following data were evaluated: imaging characteristics (length of involvement, pattern of mural thickening, pattern of attenuation, perienteric abnormalities), clinical symptoms, endoscopic records, laboratory examinations, and pathologic findings. Results Based on CT findings, two patients demonstrated segmental bowel wall thickening (involvement length >6 cm), asymmetric thickening, layered attenuation, fat stranding, and adenopathy, whereas the remaining one had no positive finding. The endoscopic results of all patients showed numerous irregular ulcers in the colon, and one patient had a focal esophageal ulcer. The major clinical symptoms were abdominal pain (n = 3), retrosternal pain (n = 1), fever (n = 3), diarrhea (n = 2), hematochezia (n = 1), and adenopathy (n = 3). The main laboratory examination indicators were increased serum EBV DNA load (n = 1) and increased inflammatory markers (n = 3). With regard to the main pathologic findings, all patients showed positive EBV-encoded RNA (EBER) situ hybridization in the colonic biopsy specimen, with one patient being positive in the esophagus. Conclusion CAEAE is rare and is usually misdiagnosed as inflammatory bowel disease (IBD). The imaging features of CAEAE overlap with those of Crohn's disease and ulcerative colitis. The presence of segmental and asymmetric bowel wall thickening, layered attenuation, and fat stranding in the CTE image may be helpful in differentiating CAEAE from IBD.

Risk factors for Epstein Barr virus-associated cancers: a systematic review, critical appraisal, and mapping of the epidemiological evidence.

BackgroundEpstein Barr Virus (EBV) infects 90%-95% of all adults globally and causes ~ 1% of all cancers. Differing proportions of Burkitt’s lymphoma (BL), gastric carcinoma (GC), Hodgkin’s lymphoma (HL) and nasopharyngeal carcinoma (NPC) are associated with EBV. We sought to systematically review the global epidemiological evidence for risk factors that (in addition to EBV) contribute to the development of the EBV-associated forms of these cancers, assess the quality of the evidence, and compare and contrast the cancers.MethodsMEDLINE, Embase and Web of Science were searched for studies of risk factors for EBV-associated BL, GC, HL and NPC without language or temporal restrictions. Studies were excluded if there was no cancer-free comparator group or where analyses of risk factors were inadequately documented. After screening and reference list searching, data were extracted into standardised spreadsheets and quality assessed. Due to heterogeneity, a narrative synthesis was undertaken.Results9916 hits were retrieved. 271 papers were retained: two BL, 24 HL, one GC and 244 NPC. The majority of studies were from China, North America and Western Europe. Risk factors were categorised as dietary, environmental/non-dietary, human genetic, and infection and clinical. Anti-EBV antibody load was associated with EBV-associated GC and BL. Although the evidence could be inconsistent, HLA-A alleles, smoking, infectious mononucleosis and potentially other infections were risk factors for EBV-associated HL. Rancid dairy products; anti-EBV antibody and EBV DNA load; history of chronic ear, nose and/or throat conditions; herbal medicine use; family history; and human genetics were risk factors for NPC. Fresh fruit and vegetable and tea consumption may be protective against NPC.ConclusionsMany epidemiological studies of risk factors in addition to EBV for the EBV-associated forms of BL, GC, HL and NPC have been undertaken, but there is a dearth of evidence for GC and BL. Available evidence is of variable quality. The aetiology of EBV-associated cancers likely results from a complex intersection of genetic, clinical, environmental and dietary factors, which is difficult to assess with observational studies. Large, carefully designed, studies need to be strategically undertaken to harmonise and clarify the evidence.RegistrationPROSPERO CRD42017059806.

T-cell posttransplant lymphoproliferative disorders after allogeneic hematopoietic cell transplantation.

Posttransplant lymphoproliferative disorder (PTLD) after allogeneic hematopoietic cell transplantation (HCT) is usually donor derived, associated with Epstein-Barr virus (EBV), and of B-cell origin. T-cell PTLD (T-PTLD) after allogeneic HCT is extremely rare. Four of 1015 (0.39%) allogeneic HCT patients were diagnosed with T-PTLD; peripheral T-cell lymphoma-not otherwise specified, anaplastic large cell lymphoma, monomorphic T-cell PTLD and polymorphic PTLD with chronic active EBV infection-like symptoms. Three of the four patients developed T-PTLD within 6months after HCT from HLA-mismatched unrelated donor. Three (75%) and 4 (100%) cases were positive for EBV-encoded small RNA in situ hybridization and EBV-DNA load in peripheral blood, respectively. Chimerism analysis showed that 75% of T-PTLD tissues (3/4) were recipient derived. T-PTLD was refractory to salvage chemotherapy and fatal in all four patients. Including the 10 patients in the literature, the median interval from HCT to diagnosis of T-PTLD was 5months (range 1-72months), 55% were negative for EBV, and 56% were recipient-derived. T-PTLD, which often occurred early after allogeneic HCT, was more likely to be EBV negative and recipient derived than B-cell PTLD after allogeneic HCT. Like T-PTLD after solid organ transplant, T-PTLD after allogeneic HCT demonstrated morphological heterogeneity and poor prognosis.

The aim of the measurement of Epstein-Barr virus DNA in hydroa vacciniforme and hypersensitivity to mosquito bites.

Epstein-Barr virus (EBV) DNA load in the blood increases in posttransplant lymphoproliferative disorders and chronic active EBV infection. In this report, we analyzed the EBV DNA load in the peripheral blood mononuclear cells (PBMCs) and plasma of patients with hydroa vacciniforme (HV) and/or hypersensitivity to mosquito bites (HMB) to understand the clinical significance of EBV DNA load. All 30 patients showed high DNA loads in the PBMCs over the cut-off level. Of 16 plasma samples, extremely high in two samples obtained from patients with hemophagocytic lymphohistiocytosis (HLH). The amount of cell-free DNA in plasma was correlated to the serum levels of lactate dehydrogenase and inversely correlated to platelet counts. These results indicate that the EBV DNA load in PBMCs can provide one of the diagnostic indicators for HV and HMB and marked elevation of cell-free EBV DNA in plasma might be related to cytolysis such as that observed in HLH.

Clinical Implications of Plasma Epstein-Barr Virus DNA in Children and Adolescent Nasopharyngeal Carcinoma Patients Receiving Intensity-Modulated Radiotherapy.

Background: Plasma Epstein–Barr virus (EBV) DNA has been determined as a prognostic factor in adult nasopharyngeal carcinoma (NPC) patients. This study was designed to evaluate the prognostic value of plasma pretreatment EBV DNA in children and adolescent NPC patients receiving intensity-modulated radiotherapy (IMRT).Methods: Pretreatment EBV DNA was retrospectively assessed in 147 children with newly diagnosed, non-metastatic NPC. All patients were treated using IMRT. Receiver operating characteristic (ROC) curve was used to identify the optimal EBV DNA cutoff point. Prognostic value was examined using a multivariate Cox proportional hazards model.Results: The median follow-up for the entire cohort was 58 months (range, 10–119 months), and the 5-year survival rates for all patients were as follows: overall survival (OS), 88.7%; locoregional relapse-free survival, 95.2%; distant metastasis-free survival (DMFS), 84.8%; and disease-free survival (DFS), 81.5%. For ROC curve analysis, the optimal cutoff value of pretreatment EBV DNA load for DFS was 40,000 copies/mL. High plasma EBV DNA was significantly associated with poorer 5-year DMFS (70.6 vs. 89.1%, P = 0.003) and DFS (63.9 vs. 86.9%, P < 0.001). In multivariate analysis, high plasma EBV DNA was an independent predictor for DMFS and DFS.Conclusions: Pretreatment EBV DNA level was a powerful prognostic discriminator for DMFS and DFS in children and adolescent NPC patients treated with IMRT.

Detection of methylation status of Epstein-Barr virus DNA C promoter in the diagnosis of nasopharyngeal carcinoma.

The detection of Epstein‐Barr virus (EBV) DNA load in nasopharyngeal (NP) brushing samples for diagnosis of nasopharyngeal carcinoma (NPC) has attracted great attention. Further improvements that eliminate the need for clinical settings will greatly extend its application. A total of 250 participants were recruited to obtain NP brushing samples. Brush sampling both with and without the guide of endoscopy was conducted in 38 NPC patients. EBV DNA load, EBV RNA transcript and EBV DNA C promoter methylation status were, respectively, evaluated. Typical latency II transcripts were observed in brushing samples from NPC patients but not controls. Unlike in tissues, multiple lytic gene transcripts were observed not only in NPC patients but also in controls. Apart from EBV RNA transcript, samples from NPC patients also showed higher levels of EBV DNA load and C promoter methylation degree than their controls. Qualitative analysis further showed that EBV DNA C promoter was methylated in all NPC patients but in only 18.4% of the control group. Combined analysis of EBV DNA methylated degree and EBV DNA load increased the sensitivity to 100% in the detection of NPC. Using qualitative methylated type as the criteria, up to 89.5% of samples collected via blind brushing showed consistent results with samples collected via endoscopy‐guided brushing from NPC patients. Detection of the methylation status of EBV DNA C promoter in NP brushing samples shows great potential in diagnosing NPC and may provide an appealing alternative for the non–invasive detection and screening of NPC without the need for clinical settings.

High viral loads of circulating Epstein-Barr virus DNA copy number in peripheral blood is associated with inferior prognosis in patients with mantle cell lymphoma.

Mantle cell lymphoma (MCL) is a distinct subtype of B cell non-Hodgkin lymphoma. No research has yet documented to investigate the prognostic implications of Epstein-Barr virus (EBV) infection in MCL. The objective of this study was to determine whether EBV DNA load may influence the heterogeneity in the course of the disease in MCL patients. Eighty-eight MCL patients were retrospectively enrolled in the study. EBV DNA load was detected by real-time quantitative PCR for quantification. The univariate and multivariate Cox proportional hazards models were established for the estimation of prognostic factors. Twenty-seven patients were detected positive for EBV DNA and the median virus titer was 1.72×104 copies/mL (range, 8.20×102 to 4.14×105 copies/mL). With a median follow-up of 39 months (range, 9 to 120 months), patients in EBV DNA-positive group displayed unfavorable progression-free survival (PFS) (P=0.012) and overall survival (OS) (P=0.004) than patients in EBV DNA-negative group. Multivariate Cox regression analysis revealed that EBV DNA-positivity was an independent risk factor for both PFS (HR, 2.04; 95% CI, 1.07 to 3.92; P=0.031) and OS (HR, 2.68; 95% CI, 1.20 to 6.00; P=0.016). Reduction in EBV copies was significantly associated with therapy-response.Circulating EBV DNA load in whole blood proved to be a significant predictor of prognosis in patients with MCL, which needs further validation in large-scale clinical studies.