Epstein-Barr Virus DNA Load Research Articles

Epstein-Barr DNA in advanced pediatric nasopharyngeal cancer

Background Studies suggest that the most common type of nasopharyngeal carcinoma (NPC) is WHO-3, which is strongly associated with Epstein-Barr virus (EBV).
 Objective To assess NPC patient characteristics in a national general referral hospital in Indonesia, with regards to EBV DNA load and treatment response.
 Methods Twenty-three pediatric patients diagnosed with NPC were included in the study. Data collected were history, physical examination, tissue biopsy, CT scan, staging and EBV DNA load from nasopharyngeal (NP) brushing as well as blood specimens. The NP brushing, blood specimens and CT scan evaluations were done two months post-treatment.
 Results Pediatric patients with symptoms such as blood tinged secretion, lymph node enlargement, and nasal congestion were more likely to have higher EBV DNA loads in their NP brushings (P<0.05) (including T3 and higher). Despite significant reduction of EBV DNA load in NP brushing post-treatment, it was not associated with treatment response, as evaluated by CT scan.
 Conclusion Higher DNA load from NP brushings is associated with a higher tumor stage. Larger sample size and follow-up data are needed to assess the usefulness of EBV DNA load assessment in pediatric patients.

The application of quantitative analysis of EBV DNA loads in plasma and peripheral blood mononuclear cells in Epstein-Barr Virus infection-related diseases in children

Objective: To explore the correlation of EBV DNA load in two different types of plasma and peripheral blood mononuclear cells (PBMCs) in children with Epstein-Barr Virus (EBV) infection diseases. Methods: A retrospective evaluated was performed on EBV DNA quantification in plasma and PBMCs by qPCR between April, 2019 and December, 2020. The samples were collected from children of 456 cases with EBV infection and 2 306 healthy cases. In EBV infection group, boys were 253 and girls were 203, aged from 8 days to months to 16 years. In healthy group, boys were 1 267 and girls were 1 039, aged from 8 days to 16 years. Results: Infectious mononucleosis (IM) was the most common disease associated with EBV infection 73.68%(336/456). The detection rate of plasma and PBMCs in EBV infection group was 91.89% (419/456)and 99.34% (453/456)respectively, and was 100%(456/456) in plasma or PBMCs. The detection rate of plasma and PBMCs in healthy group was 1.13%(26/2 306) and 30.01%(715/2 306), respectively. Levels of EBV DNA in plasma and PBMCs in EBV infection group [IM, acute infections, pneumonia, post-transplantation lymphoproliferative disorder (PTLD), hemophagocytic lymphohistiocytosis, tonsillitis and lymphadenitis] was significantly higher than those in healthy group (In plasma, Z=-47.18,-34.41,-33.40,-31.71,-24.38,-20.86 and -20.59,respectively; In PBMCs, Z=-33.17,-16.45,-11.33,-9.45,-5.57,-5.16 and -5.45, respectively; P<0.05). In IM group, EBV DNA load in plasma and PBMCs in remission stage was significantly lower than those in infection stage (Z=-11.45, -8.53;P<0.05). In PTLD group, there was significant difference in EBV DNA load in plasma between infection and remission stage (Z=-4.13, P<0.05), while there was no significant difference in EBV DNA load in PBMCs (Z=-0.817, P>0.05). Conclusions: EBV infection was mainly caused by IM. Combined detection of plasma and PBMCs in EBV DNA is valuable for improving diagnosis ability of EBV infection-related diseases, and the load of EBV DNA could be used as a marker.

Benefit of [18F]-FDG PET/CT for treatment-na\xefve nasopharyngeal carcinoma

BackgroundTo test the advantages of positron emission tomography and computed tomography (PET/CT) for diagnosing lymph nodes and staging nasopharyngeal carcinoma and to investigate its benefits for survival and treatment decisions.MethodsThe performance of PET/CT and magnetic resonance imaging (MRI) in diagnosis was compared based on 460 biopsied lymph nodes. Using the propensity matching method, survival differences of T3N1M0 patients with (n = 1093) and without (n = 1377) PET/CT were compared in diverse manners. A radiologic score model was developed and tested in a subset of T3N1M0 patients.ResultsPET/CT performed better than MRI with higher sensitivity, accuracy, and area under the receiver operating characteristic curve (96.7% vs. 88.5%, p < 0.001; 88.0% vs. 81.1%, p < 0.001; 0.863 vs. 0.796, p < 0.05) in diagnosing lymph nodes. Accordingly, MRI-staged T3N0-3M0 patients showed nondifferent survival rates, as they were the same T3N1M0 if staged by PET/CT. In addition, patients staged by PET/CT and MRI showed higher survival rates than those staged by MRI alone (p < 0.05), regardless of the Epstein-Barr virus DNA load. Interestingly, SUVmax-N, nodal necrosis, and extranodal extension were highly predictive of survival. The radiologic score model based on these factors performed well in risk stratification with a C-index of 0.72. Finally, induction chemotherapy showed an added benefit (p = 0.006) for the high-risk patients selected by the model but not for those without risk stratification (p = 0.78).ConclusionPET/CT showed advantages in staging nasopharyngeal carcinoma due to a more accurate diagnosis of lymph nodes and this contributed to a survival benefit. PET/CT combined with MRI provided prognostic factors that could identify high-risk patients and guide individualized treatment.

Overall Status of Epstein-Barr virus Infection, IFN-a, and TLR-7/9 in Patients with Systemic Lupus Erythematous.

Systemic lupus erythematous (SLE) is a multisystem autoimmune disorder. While studying the pathogenesis of SLE is prevalent, both infectious and non-infectious elements are regarded to exert an important impact on the disease's development. To explore the overall status of EBV, TLR7, TLR9, and IFN-α gene expression in 32 patients suffering from SLE and 32 healthy controls. Plasma and PBMCs were separated from fresh whole blood. To measure EBV DNA load and mRNA levels of IFN-a, TLR-7 and9 in PBMCs, molecular techniques were employed. The production of IFN-α, ds-DNA IgG antibody, and EBNA-1 IgG levels were also measured in plasma by ELISA. SLE patients showed significantly higher EBV load (p=0.001) and transcriptional levels of TLR7 (p=0.0001), IFN-α (p=0.0001), and TLR9 (p=0.0001) than controls. Moreover, the plasma levels of IFN-α (p=0.0002) and EBNA-1specific IgG antibodies (p=0.01) were significantly higher in SLE patients. The results stressed on the potential role of EBV infection and TLRs in SLE patients although more research is needed to determine the global impact that EBV infection can have on immune signature in patients with SLE.

Risk factors and regularity of transaminase elevation associated with primary Epstein-Barr virus infection in children

Objective: To explore the risk factors and regularity of pediatric primary Epstein-Barr virus (EBV) infection accompanied with elevated transaminase. Methods: Clinical data of 399 children diagnosed as primary EBV infection in the outpatient department, Children's Hospital of Fudan University from September 2016 to October 2017 were analyzed retrospectively. Logistic regression analysis was performed to determine the potential correlations between elevated alanine transaminase (ALT) or aspartate transaminase (AST) and age, gender, course of fever and plasma EBV-DNA load. The cumulative rates of elevated transaminase recovery to nomal at different times were caculated. Results: Among 399 children diagnosed with primary EBV infection, there were 219 males and 180 females. The age was (4.2±2.7) years. Among all cases, 51.9% (207/399) had elevated transaminase. In patients who had elevated ALT, 74.5% (149/200), 21.0% (42/200) and 4.5% (9/200) had mild (40-160 U/L), moderate (160-400 U/L) and severe (>400 U/L) elevation of ALT, respectively. In patients who had elevated AST, 83.8% (155/185), 11.9% (22/185) and 4.3% (8/185) had mild (40-160 U/L), moderate (160-400 U/L) and severe (>400 U/L) elevation of AST, respectively. Only age was correlated with the occurrence of elevated transaminase (OR=1.13, 1.10, both P<0.05). A total of 167 repeated tests were ordered in patients with elevated ALT and/or AST, including 113 cases with elevated ALT and 104 cases with elevated AST. The time of ALT and AST returned to normal were (24±13) days and (25±18) days respectively. The cumulative rates for ALT returned to normal within 1, 1-<4, 4-<8 weeks and more than 8 weeks were 2.7% (3/113), 54.0% (61/113), 79.6% (90/113) and 81.4% (92/113) respectively, and were 1.9% (2/104), 48.1% (50/104), 71.2% (74/104) and 74.0% (77/104) for AST. Conclusions: Age is a risk factor for transaminase elevation associated with primary EBV infection in children. The transaminases returned to normal within 3 weeks in half of the cases, and within 8 weeks in most cases.

Inverse Correlation of Vδ2-T-Cell Recovery with EBV Reactivation after Haploidentical Hematopoietic Stem Cell Transplantation

Background. Epstein-Barr virus (EBV) reactivation remains one of the life-threatening complications in recipients undergone haploidentical hematopoietic stem cell transplantation (haploHSCT). Reconstitution of traditional adaptive T lymphocytes is generally compromised at the early stage following transplantation, suggesting an important role of other immune effector cells in preventing from EBV reactivation. Our previous studies demonstrated that hampered recovery of CD4-CD8- T cells was associated with an increased occurrence of EBV reactivation after haploHSCT. γδ T is a dominant subgroup of CD4-CD8- T cells in the peripheral blood of healthy donors. Correlation of reconstituted γδ T cells with EBV reactivation has not been reported in the context of allogeneic HSCT.Methods. This study prospectively included 132 adult patients with hematopoietic malignancy and receiving haploHSCT. Recovery characteristics of T-cell subpopulations were monitored by flow cytometry at 30, 60, and 90 days after haploHSCT. EBV-DNA loads in the peripheral blood were detected by real-time quantitative PCR before and after transplantation. Association between γδ T recovery and EBV reactivation were statistically analyzed.Results. The median counts of peripheral Vδ2 cells were continuously lower in recipients with EBV reactivation compared with controls at 30, 60, and 90 days post-haploHSCT (P value was 0.006, <0.001, and 0.019, respectively). Whereas no significant association of Vδ1-cell recovery was found in the same setting. Landmark study further indicated that the cumulative incidence of EBV reactivation was significantly increased in recipients with lower day-30 Vδ2 cell counts. Cytotoxic effect of Vδ2 cells on EBV-targeted cells were further confirmed by in-vitro experiments. Consistently, recovered Vδ2-positive in contrast to Vδ2-negative γδ T cells were specifically activated upon EBV reactivation following haploHSCT. The incidences of CMV reactivation, bacteremia, invasive fungal infection (IFI), and II-IV and III-IV aGVHD were not significantly different between the groups with and without EBV reactivation (P > 0.1, respectively).Conclusions. We clarified a specific association of recovered Vδ2 but not Vδ1 T cells with EBV reactivation following haploHSCT. These results may help better understand the intrinsic anti-EBV immunity after allogeneic HSCT and develop effective γδ T-cell-based therapy strategies.AcknowledgmentsThis study is supported by the Foundation for Innovative Research Groups of the National Natural Science Foundation of China (Grant No. 81621001) and National Natural Science Foundation of China (Grant No. 81370666). DisclosuresNo relevant conflicts of interest to declare.

Chidamide ,Oral Subtype-Selective Histone Deacetylase Inhibitor (HDACI) Monotherapy Was Effective on the Patients with Relapsed or Refractory Extranodal Natural Killer (NK)/T-Cell Lymphoma

Background:Chidamide is an orally bioavailable benzamide class of histone deacetylase inhibitor (HDACi), that selectively inhibits activity of HDAC1, 2, 3 and 10. It was approved by the Chinese FDA for relapsed or refractory (R/R) peripheral T-cell lymphoma (PTCL) with a recommended dose of 30 mg twice weekly (BIW). However, the efficacy and safety of Chidamide remain to be determined in relapsed or refractory extranodal natural killer T-cell lymphoma (ENKTCL), and the optimal dosing schedule was unclear. The purpose of the current study were to evaluate the clinical response and toxicity for R/R-ENKTCL at two dosing schedule of Chidamide monotherapy.Methods:This is an open-label, prospective phase II trial in China. Inclusion criteria included histologically confirmed ENKTCL, disease progression or non-response after asparaginase-based chemotherapy. Chidamide 30mg BIW or 10mg daily was taken at least 6 weeks until disease progression or the occurrence of unacceptable toxicity. Utility of PET-CT in the therapeutic evaluation was employed. Serum EBV-DNA was monitored weekly. At the time of this abstract submission, the trial is still ongoing.Results: A total of 15 relapsed or refractory ENKTCL patients failed to asparaginase-based chemotherapy or autologous hematopoietic stem cell transplantation (ASCT) were enrolled in this study (10mg QD group, n=9 and 30mg BIW group, n=6, respectively). Median previous chemotherapy regimens was 3(2-5), 2 patient relapsed following ASCT. 12 patients were evaluable for response and 3 patients still under treatment less than 6 weeks. ORR was 50.0% (6/12) with CR rate 33.3 %(4/12) . Three patients got CR in 30mg group, one patient got CR in 10mg group (table1,figure 1). The median time to response was 5.9 (1.1 to 8.1) weeks. Median follow-up time was 3.7 months (0.8-11.5) so far. All four CR patients were still in disease-free more than 6.9 months (6.9-10.5). Serum EBV-DNA load during the course of treatment was declined or stable in all responders only two cases with elevated EBV-DNA level observed due to disease progression (figure 2). Most adverse events (AEs) were grade 1 or 2, and AEs ≥grade 3 hematologic toxicity were similar in both groups included neutropenia (50.0% vs. 44.4%) and thrombocytopenia (33.3% vs. 33.3%). 30mg group experienced a higher incidence of nausea and vomiting(100% vs. 44.4%). Meanwhile, we also treated six refractory B cell lymphoma patients, including three diffused large B cell lymphoma (DLBCL), two mantle cell lymphoma (MCL) and one follicular lymphoma. One DLBCL patient and one MCL patient obtained PR respectively.Conclusions: Our study suggested that Chidamide was effective as single agent in the treatment of relapsed or refractory malignant lymphoma.. The therapeutic outcome for previous heavily-treated ENKTCL was encouraged without clinical EBV reactivation. It is too early to draw conclusions of response difference between two dosing schedule. Further clinical trial warranted.(ClinicalTrials.gov, NCT 2878278 )Disclosures: No relevant conflicts of interest to declare. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Establishment of a Prognostic Nomogram for Patients With Locoregionally Advanced Nasopharyngeal Carcinoma Incorporating TNM Stage, Post-Induction Chemotherapy Tumor Volume and Epstein-Barr Virus DNA Load

ObjectivesTo establish and validate an effective nomogram to predict clinical outcomes for patients with locoregionally advanced nasopharyngeal carcinoma (LA-NPC).Materials and MethodsThe clinicopathological parameters and follow-up information of 402 locoregionally advanced NPC patients (training cohort, n = 302; validation cohort, n = 100) were retrospectively enrolled. The nomogram was built with the important prognostic variables identified by Cox regression analysis. Overall survival (OS) and progression-free survival (PFS) were the primary and secondary endpoints, respectively. The predictive power and clinical utility of the nomogram were assessed using the Harrell concordance index (C-index), calibration curve, and decision curve analysis. We compared the eighth staging system model with the nomogram to analyze whether the model could improve the accuracy of prognosisResultsEpstein–Barr virus (EBV) DNA load, the gross tumor volume (GTVnx), and cervical lymph node tumor volume (GTVnd) after induction chemotherapy were the independent predictors of OS and PFS. The calibration curves indicated superb agreement between the nomogram-predicted probabilities and observed actual probabilities of survival. The C-index and area under the receiver operator characteristic curve (AUC) of the nomogram integrating these significant factors and N stage, and TNM stage were higher than those of the eighth TNM system alone. In addition, the decision curve analyses demonstrated the clinical value and higher overall net benefit of the nomogram. High-risk groups identified by the nomogram had significantly poorer OS and PFS than the low-risk group (p < 0.05).ConclusionsThe multidimensional nomogram incorporating TNM stage, EBV DNA load, and tumor volume after induction chemotherapy led to a more precise prognostic prediction and could be helpful for stratifying risk and guiding treatment decisions in locoregionally advanced NPC patients who have undergone induction chemotherapy and concurrent chemoradiation.

PLASMA EPSTEIN-BARR VIRUS (EBV) DNA AS A BIOMARKER FOR DIAGNOSIS OF SYRIAN EBV-POSITIVE BURKITT’S LYMPHOMA

Epstein-Barr virus - positive Burkitt’s Lymphoma is defined as the presence of Epstein-Barr virus (EBV) in tumor cells, the standard way to detect (EBV) in Burkitt’s Lymphoma is in-situ hybridization (ISH) of EBV-encoded small RNA (EBERs) in tumor cells. The present study aimed to evaluate plasma Epstein-Barr virus (EBV) DNA as a noninvasive biomarker for diagnosis and prognosis of EBV-positive Burkitt’s lymphoma. The study included 40 newly diagnosed patients with Burkitt’s lymphoma, ranging in age from 4 to 60 years, and 55 sex and age-matched controls. Forty formalin-fixed paraffin-embedded blocks of Burkitt’s lymphoma tissue samples were used to investigate the EBV by in-situ hybridization detection of the EBERs. Plasma EBV DNA was quantified by real-time quantitative polymerase chain reaction (PCR) for all Burkitt lymphoma patients prior to therapy and for control. The results showed that (22/40, 55%) of Burkitt lymphoma were positive for histological EBER, whereas plasma EBV DNA was detectable (range from 1.2×104 to 4.7×106 copies/mL) in all EBV-positive Burkitt lymphoma samples (22/22). EBV DNA was undetectable in all cases of EBV-negative Burkitt lymphomas (18/18) and all healthy control (55/55). It is worth mentioning that our results demonstrated that the EBV DNA load was significantly high in the EBV-positive BL patients suffering poor prognostic state. In conclusion: Plasma EBV-DNA can be used as a noninvasive biomarker for diagnosis and prognosis of EBV-positive Burkitt’s lymphoma.

Plasma Epstein-Barr virus DNA and risk of nasopharyngeal carcinoma in a prospective seropositive population

ObjectivePlasma Epstein-Barr virus (EBV) DNA is considered a biomarker for nasopharyngeal carcinoma (NPC). However, its long-term role in NPC development is unclear.Materials and methodsA total of 1363 participants seropositive for EBV VCA-IgA and EBNA1-IgA in a community-based NPC screening program in southern China were tested for plasma EBV DNA levels by real-time qPCR between 2008 and 2015. New NPC cases were confirmed by active follow-up approach and linkage to local cancer registry through the end of 2016. Cox proportional hazards regression analysis was performed to calculate the hazard ratios (HRs) for NPC risk with plasma EBV DNA.ResultsThirty patients were newly diagnosed during a median 7.5 years follow-up. NPC incidence increased with the plasma EBV DNA load ranging from 281.46 to 10,074.47 per 100,000 person-years in participants with undetectable and ≥ 1000 copies/ml levels; the corresponding cumulative incidence rates were 1.73 and 50%. Furthermore, plasma EBV DNA loads conferred an independent risk for NPC development after adjustment for other risk factors, with HRs of 7.63 for > 3–999 copies/ml and 39.79 for ≥1000 copies/ml. However, the HRs decreased gradually after excluding NPC cases detected in the first 2 to 3 years and became statistically nonsignificant by excluding cases detected during the first 4 years.ConclusionElevated plasma EBV DNA can predict NPC risk over 3 years. Monitoring plasma EBV DNA can be used as a complementary approach to EBV serological antibody-based screening for NPC.

Hydroa vacciniforme‐like lymphoproliferative disorder: A clinicopathological, immunohistochemical, and prognostic study of 24 cases in China

Hydroa vacciniforme‐like lymphoproliferative disorder (HV‐LPD) is a rare cutaneous disease associated with Epstein–Barr virus infection. We retrospectively analyzed the clinical presentation, histopathological characteristics, and prognostic study of HV‐LPD in 24 Chinese patients. All patients presented with recurrent papulovesicular and necrotic eruptions on the face, neck, and extremities, with 11 showing systemic symptoms. Twenty patients were diagnosed with HV‐LPD in childhood (age < 18 years) and four in adulthood (age ≥ 18 years). The median age at diagnosis was 8.5 years old (range, 2–50). Histopathology revealed variably dense lymphocyte infiltration throughout the dermis. All cases were strongly positive for CD3 and Epstein–Barr encoding region based on in situ hybridization. Of 18 cases with a T‐cell phenotype, 15 harbored monoclonal rearrangements in T‐cell receptor (TCR) genes. Four cases with a natural killer cell phenotype carried polyclonal rearrangements in TCR genes. Among 24 patients, eight (33.3%) received chemotherapy, two (8.3%) allogeneic hematopoietic stem cell transplantation, and both are currently alive without disease. The median follow‐up period was 24 months (range, 7–120) and 23 patients were available: 15 (62.5%) were alive, and eight (33.3%) had died. Fourteen cases had a relapse of disease and three developed lymphoma within 24 months of diagnosis. The mean survival time of childhood‐onset patients was longer than that of adult‐onset patients (36.4 vs. 20.8 months). In summary, the wide clinical course and representative presentation of cases in our center reflect the pedigree characteristics of HV‐LPD. Allogeneic hematopoietic stem cell transplantation should be a preferred choice for relapse and refractory patients due to the poor effect of chemotherapy. Adult‐onset and high serum EBV DNA loads may indicate an increased risk of aggressive disease in patients with HV‐LPD.

Innate Immune Modulation Induced by EBV Lytic Infection Promotes Endothelial Cell Inflammation and Vascular Injury in Scleroderma

Microvascular injury is considered an initial event in the pathogenesis of scleroderma and endothelial cells are suspected of being the target of the autoimmune process seen in the disease. EBV has long been proposed as a trigger for autoimmune diseases, including scleroderma. Nevertheless, its contribution to the pathogenic process remains poorly understood. In this study, we report that EBV lytic antigens are detected in scleroderma dermal vessels, suggesting that endothelial cells might represent a target for EBV infection in scleroderma skin. We show that EBV DNA load is remarkably increased in peripheral blood, plasma and circulating monocytes from scleroderma patients compared to healthy EBV carriers, and that monocytes represent the prominent subsets of EBV-infected cells in scleroderma. Given that monocytes have the capacity to adhere to the endothelium, we then investigated whether monocyte-associated EBV could infect primary human endothelial cells. We demonstrated that endothelial cells are infectable by EBV, using human monocytes bound to recombinant EBV as a shuttle, even though cell-free virus failed to infect them. We show that EBV induces activation of TLR9 innate immune response and markers of vascular injury in infected endothelial cells and that up-regulation is associated with the expression of EBV lytic genes in infected cells. EBV innate immune modulation suggests a novel mechanism mediating inflammation, by which EBV triggers endothelial cell and vascular injury in scleroderma. In addition, our data point to up-regulation of EBV DNA loads as potential biomarker in developing vasculopathy in scleroderma. These findings provide the framework for the development of novel therapeutic interventions to shift the scleroderma treatment paradigm towards antiviral therapies.

Antineoplastic and anti-inflammatory effects of bortezomib on systemic chronic active EBV infection

Systemic chronic active Epstein-Barr virus (EBV; sCAEBV) infection, T- and natural killer (NK)-cell type (sCAEBV), is a fatal disorder accompanied by persisting inflammation harboring clonal proliferation of EBV-infected T or NK cells. Today's chemotherapy is insufficient to resolve disease activity and to rid infected cells of sCAEBV. The currently established treatment strategy for eradicating infected cells is allogeneic hematopoietic stem cell transplantation. In this study, we focused on the effects of proteasome inhibitor bortezomib on the disease. Bortezomib suppressed survival and induced apoptosis of EBV+ T- or NK-cell lines and peripheral mononuclear cells containing EBV-infected T or NK cells of sCAEBV patients. Bortezomib enhanced binding immunoglobulin protein/78-kDa glucose-regulated protein (Bip/GRP78) expression induced by endoplasmic reticulum stress and activated apoptosis-promoting molecules JNK and p38 in the cell lines. Bortezomib suppressed the activation of survival-promoting molecule NF-κB, which was constitutively activated in EBV+ T- or NK-cell lines. Furthermore, quantitative reverse transcription–polymerase chain reaction demonstrated that bortezomib suppressed messenger RNA expression of proinflammatory cytokines tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ) in EBV+ T or NK cells from the patients. Finally, we examined the effects of bortezomib using xenograft models of sCAEBV generated by IV injection of patients' cells. The intraperitoneal administration of bortezomib significantly reduced EBV-DNA load in peripheral blood and the infiltration of EBV-infected cells in the models' livers. Moreover, the serum concentration of TNF-α and IFN-γ decreased after bortezomib treatment to the models. Our findings will be translated into the treatment of sCAEBV not only to reduce the number of tumor cells but also to suppress inflammation.

Circulating cell\u2010free epstein\u2013barr virus DNA levels and clinical features in Moroccan patients with nasopharyngeal carcinoma

BackgroundThe identification of effective prognosis biomarkers for nasopharyngeal carcinoma (NPC) is crucial to improve treatment and patient outcomes. In the present study, we have attempted to evaluate the correlation between pre-treatment plasmatic Epstein-Barr virus (EBV) DNA load and the conventional prognostic factors in Moroccan patients with NPC.MethodsThe present study was conducted on 121 histologically confirmed NPC patients, recruited from January 2017 to December 2018. Circulating levels of EBV DNA were measured before therapy initiation using real-time quantitative PCR.ResultsOverall, undifferentiated non-keratinizingcarcinoma type was the most common histological type (90.1 %), and 61.8 % of patients were diagnosed at an advanced disease stage (IV). Results of pre-treatment plasma EBV load showed that 90.9 % of patients had detectable EBV DNA, with a median plasmatic viral load of 7710 IU/ml. The correlation between pre-treatment EBV DNA load and the conventional prognostic factors showed a significant association with patients’ age (p = 0.01), tumor classification (p = 0.01), lymph node status (p = 0.003), metastasis status (p = 0.00) and overall cancer stage (p = 0.01). Unexpectedly, a significant higher level of pre-treatment EBV DNA was also found in plasma of NPC patients with a family history of cancer (p = 0.04). The risk of NPC mortality in patients with high pretreatment EBVDNA levels was significantly higher than that of those with low pre-treatment plasma EBV-DNA levels (p < 0.05). Furthermore, patients with high pre-treatment EBV-DNA levels (≥ 2000, ≥ 4000) had a significant low overall survival (OS) rates (p < 0.05). Interestingly, lymph node involvement, metastasis status and OS were found to be the most important factors influencing the EBV DNA load in NPC patients.ConclusionsThe results of the present study clearly showed a high association between pre-treatment EBV DNA load, the crucial classical prognostic factors (T, N, M and disease stage) of NPC and OS, suggesting that pre-treatment EBV DNA can be a useful prognostic biomarker in clinical decision-making and improving NPC treatment in Morocco.

Prospective evaluation of blood Epstein-Barr virus DNA load and antibody profile in HIV-related non-Hodgkin lymphomas.

The value of Epstein-Barr virus (EBV) biomarkers on the prognosis of HIV-related non-Hodgkin's lymphoma (NHL) has been poorly explored in the combined antiretroviral therapy (cART) era. We evaluated EBV DNA load and EBV antibodies in HIV-NHL patients enrolled in the French ANRS-CO16 Lymphovir Cohort between 2008 and 2015. Whole blood and plasma EBV DNA load and serological profiles were analyzed in 76 HIV-infected patients at diagnosis of NHL and 6 months after the initiation of chemotherapy. Prechemotherapy whole blood (WB) and plasma EBV DNA loads were positive for 80 and 45% of HIV-NHL patients, respectively. Pretreatment WB EBV DNA positivity was associated with a positive plasma HIV-1 RNA load (relative risk (RR), 4.42 [1.33; 14.72]) and plasma EBV DNA positivity with EBV in situ detection (RR 10.62 [2.38; 47.49]). Following chemotherapy, the proportions of patients with positive WB or plasma EBV DNA declined from 81 to 23% (P < 0.0001) and from 43 to 8% (P < 0.0001), respectively. Estimated 2-year progression-free survival did not differ according to prechemotherapy WB positivity (82% versus 67%, P = 0.15) or plasma EBV DNA positivity (76% versus 81%, P = 0.52). The plasma EBV DNA load correlates with in situ EBV detection. The WB EBV DNA load correlates with HIV load. WB and plasma EBV DNA loads at NHL diagnosis do not constitute prognostic markers for HIV-NHL patients in the modern cART era.

Plasma Epstein-Barr Virus (EBV) DNA as a Biomarker for Diagnosis of Syrian EBV-Positive Burkitt’s Lymphoma

Epstein-Barr Virus -positive Burkitt’s Lymphoma is defined as the presence of Epstein-Barr virus (EBV) in tumor cells, the standard way to detect (EBV) in Burkitt’s Lymphoma is in situ hybridization (ISH) of EBV-encoded small RNA (EBERs) in tumor cells. The present study aimed to evaluate plasma Epstein-Barr virus (EBV) DNA as a noninvasive biomarker for diagnosis and prognosis of EBV-positive Burkitt’s lymphoma. The study included 40 newly diagnosed patients with Burkitt’s lymphoma, ranging in age from 4 to 60 years, and 55 sex and age-matched controls. (40) Formalin-fixed paraffin embedded blocks of Burkitt’s lymphoma tissue samples were used to investigate the EBV by in situ hybridization detection of the EBERs. Plasma EBV DNA was quantified by real-time quantitative polymerase chain reaction (PCR) for all Burkitt lymphoma patients prior to therapy and for control. The results showed that (22/40, 55%) of Burkitt lymphoma were positive for histological EBER, whereas plasma EBV DNA was detectable (range from 1.2 × 104 to 4.7 × 106 copies/mL) in all EBV-positive Burkitt lymphoma samples (22/22). EBV DNA was undetectable in all cases of EBV-negative Burkitt lymphomas (18/18) and all healthy control (55/55). It is worth mentioning that our results demonstrated that the EBV DNA load was significantly high in the EBV-positive BL patients suffering poor prognostic state. In conclusion: Plasma EBV-DNA can be used as a noninvasive biomarker for diagnosis and prognosis of EBV-positive Burkitt’s lymphoma.

Prognosis predicting value of semiquantitative parameters of visceral adipose tissue and subcutaneous adipose tissue of 18F-FDG PET/CT in newly diagnosed secondary hemophagocytic lymphohistiocytosis.

The purpose of this study was to investigate the prognosis predicting value of visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) of 18F-FDG PET/CT, and clinical inflammatory cytokines in newly diagnosed secondary hemophagocytic lymphohistiocytosis (SHLH). We retrospectively collected 58 patients with newly diagnosed SHLH from August 2016 to July 2019 in our hospital. All patients were followed up between 6 and 24months. First, a comprehensive comparison of the general data between the death and the survival group was performed. Clinical lab indexes included were recorded and analyzed retrospectively. Second, the correlation between 18F-FDG PET/CT semiquantitative metabolic parameters of VAT, SAT and inflammatory cytokines was performed. 3D slicer software was used to get SUV and volume of VAT and SAT from 18F-FDG PET/CT. Third, overall survival (OS) analysis was performed. Finally, the prognosis predicting model was built based on risk factors to stratify SHLH patients. There was significant difference in WBC, PLT, FBG, IL-10, PCR tests of EBV-DNA loads, SCD25 between the death group and the survival group. There was significant correlation between SAT coefficient variance (CV) and CRP, the mean standardized uptake value (SUVmean) of SAT (SAT SUVmean) and TG, SAT SUVmean and ESR. In univariate analysis with Cox regression analysis, SUVmean of VAT (VAT SUVmean), SAT Volume, SUVmean of SAT, CV of SAT (SAT HU CV), plasma EBV-DNA, WBC, PLT, FBG showed significance with OS. In multivariate Cox regression analysis, SAT Volume, SUVmean of SAT, plasma EBV-DNA, were independent prognostic factors for OS. For newly diagnosed SHLH, SAT Volume, SUVmean of SAT, plasma EBV-DNA had significant relationship with poor prognosis. They were important independent predictors for overall survival for newly diagnosed SHLH.

EBV Serology and Plasma EBV DNA Load as a Combined Tool for Nasopharyngeal Carcinoma (NPC) Diagnosis and Monitoring in Non-Endemic Regions

There is limited information regarding the clinical significance of Epstein-Barr virus (EBV) markers for nasopharyngeal carcinoma (NPC) diagnoses and monitoring in non-endemic areas. Low NPC incidence in non-endemic countries made it difficult to form a representative group of patients for research this issue. Moreover, NPC cases are often characterized into morphologically different tumor types based upon the geographic and ethnic variability. Since viral and serological markers reflect the different biological events accompanying the development of NPC, it is important to compare their clinical value in the context of different disease manifestations. The implementation of such a study in a non-endemic region is of particular interest, allowing the investigation of the potential impact of differences in the genetic and ethnic characteristics of the population, versus those in populations from endemic regions. In present study, we analyzed clinical significance of two EBV markers (serological and molecular) in large group (96 cases) of undifferentiated non-keratinizing carcinoma of nasopharyngeal type (UNPC) Russian patients. It has been shown that IgA/ VCA antibody titers elevated on patient's admission and being valuable markers for primary UNPC diagnosis do not allow to adequately assessing patients’ state after the treatment. In contrast to EBV serology, the plasma EBV DNA load was found to be valuable marker for clinical evaluation of UNPC patient’s state, such as remission and relapse. It was also shown that the concentration of viral DNA correlated with the UNPC patients' overall survival. The proposed study, conducted on UNPC patients from a non-endemic region, for the first time revealed a direct correlation between IgG/IgA antibody titers to EBV virus capsid antigen (VCA) and the levels of plasma EBV DNA load, and the absence of such correlation between plasma EBV DNA burden and serological responses to EBV. It has also been demonstrated that the combined assessment of plasma EBV DNA load and EBV-specific antibody titers provides a reliable approach to UNPC diagnosis, disease monitoring, and therapeutic response assessment.

Clinical warning of hemophagocytic syndrome caused by Epstein-Barr virus

ObjectivesThis study aimed to compare the clinical features and laboratory tests of infectious mononucleosis (IM) and hemophagocytic syndrome (HLH) caused by Epstein-Barr virus (EBV) in 1–3-year-old children and to explore the risk factor of HLH caused by EBV (EBV-HLH).MethodsThe clinical data of 92 children with EBV infection admitted in our hospital from 2011 to 2019 were collected; 61 cases were diagnosed as EBV-IM, and 31 cases were diagnosed as EBV-HLH. The subjects’ clinical manifestations and laboratory tests were analyzed retrospectively.ResultsCompared with EBV-IM patients, EBV-HLH patients had longer durations of fever, both before hospitalization and overall, and a higher probability of hepatomegaly. The levels of ALT, AST, LDH, TG, SF, D-Dimer and the plasma EBV DNA load of EBV-HLH patients were significantly higher than those of EBV-IM patients. The absolute values of CD3+, CD4+, CD8+, NK, and CD3-CD19+ cells and IgA and IgM levels of EBV-HLH patients were significantly lower than those of EBV-IM patients. The plasma EBV DNA load was positively correlated with the PT, TT, α-HBDH, AST, LDH, CK, Scr, BUN, UA, TG, and CRP levels in EBV-HLH patients, and the plasma EBV DNA load was positively correlated with the D-Dimer level in the EBV-IM patients. Among the 10 different potential markers, at the cut-off point of 1721.500 μg/L, the sensitivity and specificity of D-Dimer was 88.90 and 90.20%, respectively.ConclusionThe D-Dimer level may be a good prognostic indicator of EBV-HLH caused by EBV.

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.