Plasma EBV DNA Research Articles

Clinical Significance of Plasma Epstein-Barr Virus DNA in Peripheral T-Cell Lymphomas

Introduction: The clinical implications of plasma Epstein-Barr virus (EBV) DNA in patients with peripheral T-cell lymphoma (PTCL) remain unclear. Objective: This study aimed to explore the clinical correlations of pre- and post-treatment plasma EBV-DNA concentrations with treatment outcomes and prognosis in patients with PTCL. Methods: We retrospectively reviewed 128 patients diagnosed with PTCL with available data on pre-treatment plasma EBV-DNA, including 63 patients for whom post-treatment plasma EBV-DNA data were also available. Patients with extra-nodal NK/T-<X00_Del_TrennDivis></X00_Del_TrennDivis>cell lymphoma were excluded from this study. Results: Pre-treatment plasma EBV-DNA was elevated (e.g., ≥500 copies/mL) in 35.2% of PTCL patients, with significant differences in positive rates between different subtypes of PTCL (p < 0.001). High pre-treatment EBV-DNA concentrations were associated with advanced age (>60 years), elevated lactate dehydrogenase levels, high International Prognostic Index (IPI), and positive EBV-encoded RNAs-ISH in tumor specimens. In multivariate analyses, pre-treatment EBV-DNA ≥500 copies/mL was an independent prognostic factor after adjusting for IPI and pathological subtypes (hazard ratio = 2.14, p = 0.032). For patients with elevated pre-treatment EBV-DNA, normalization of EBV-DNA concentrations after first-line chemotherapy was significantly associated with better overall response rate (81.3% vs. 22.2%, p = 0.014), progression-free survival (12.0 months vs. 3.7 months, p = 0.011), and overall survival (37.9 months vs. 7.8 months, p = 0.012). For patients achieving remission to first-line therapy, rebound of EBV-DNA levels during follow-up was associated with disease relapse or progression. Conclusions: These results suggest that pre-treatment plasma EBV-DNA concentration is a strong prognostic factor for PTCL. For patients with elevated pre-treatment EBV-DNA, dynamic monitoring of EBV-DNA changes after initiation of therapy is useful for predicting treatment outcome and disease relapse.

Immunodeficiency Genes Are Associated with EBV Positive Malignancies and Hemophagocytic Lymphohistiocytosis That Can be Cured By Allogeneic Hematopoietic Stem Cell Transplantation

Background: Epstein-Barr virus (EBV) positive hematological malignancies and EBV-associated hemophagocytic lymphohistiocytosis (HLH) are serious illness that is related with chronic active EBV infection (CAEBV). Recently, more and more immune-related gene mutations have been identified to associate with the EBV positive hematological diseases. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curative therapy for above diseases.Aim: In present clinical study, the germline gene mutations associated with EBV positive hematological malignancies and EBV positive HLH were analyzed and the outcomes of allo-HSCT in those diseases were investigated.Methods: Between January 2018 and June 2021, 7 patients with EBV positive hematological diseases (lymphoma 3, aggressive NK-cell leukemia (ANKL) 1, secondary HLH 3) who received allo-HSCT in our hospital were included. Blood samples from patients, parents and potential related donors were collected before transplantation and whole exome sequencing of the blood DNA samples was performed to detect and analyze their relevance with the clinical features before and after allo-HSCT. The median age was 23 (5-31) years old. The median disease course before transplant was 6 (3-38) months. Before allo-HSCT, 3 patients (HLH 1, lymphoma 1, ANKL 1) were in non-remission (NR), 2 patients with lymphoma were in partial remission (PR) and the other two patients with HLH were in complete remission (CR). Five patients received allo-HSCT from haploidentical donors and 2 patients from unrelated donors (HLA 10/10, 9/10 matched). Myeloablative conditioning regimen with busulfan (0.8mg/kg iv q6h, 3 days)/etoposide (200mg/m 2, 3 days)/fludarabine (30mg/m 2, 5 days) /rabbit anti-T-cell globulin was used. Cyclosporine, mycophenolate mofetil and short-term methotrexate were employed for graft-versus-host disease (GVHD) prophylaxis.Results: All patients achieved durable engraftment. Plasma EBV-DNA was positive in three patients (1200, 880, 84000 copies/ml, respectively) before transplant and has been negative in all patients after transplant. One patient had gastrointestinal bleeding caused by EBV enteritis. Two patients had CMV viremia. Three patients developed grade III acute GVHD and two had extensive chronic GVHD. All GVHD was resolved with immune-suppressants. With the median follow-up time of 21 (7-36) months, all patients are alive and free of their diseases. Seven disease-related gene mutations were detected including FANCA p.S1088F, TGFBR2 p.T315M, IRF7 p.K446R, PRF1 p.R4C, MPEG1 p.P316S, STX11 p.R49Q, and UNC13D p.G863D. The first 5 gene mutations were detected in the patients with lymphoma and ANKL, and the last 2 gene mutations were detected in the patients with HLH. FANCA is associated with bone marrow failure and the other genes are associated with primary immunodeficiency, all of these gene mutations have been reported and affect in part the function of the genes. In particular, STX11 p.R49Q and UNC13D p.G863D have also been reported in HLH patients and the NK cell activity of these patients is lower than normal levels. Apart from the defined gene mutations in primary HLH, recently, many new immunodeficiency gene mutations have been reported to associate with EBV positive malignancies and HLH which suggests immunodeficiency might play a role in the etiology of the diseases. The gene mutations we reported here might be the main gene mutations associated with disease susceptibility and infection risk.Conclusion: Our preliminary results have shown the gene mutation profile related to EBV positive hematological malignancies and HLH. With our transplant protocol, durable reconstitution of hematopoiesis and immune from healthy donors has been achieved successfully which has resulted in 100% disease-free survival and no transplant-related death. Allo-HSCT is a safe and effective curative modality for EBV positive hematological malignancies and HLH. DisclosuresNo relevant conflicts of interest to declare.

Association of B Symptoms With Plasma EBV-DNA Copy Number and Cytokine Profiles in Patients With Nasal Type Extranodal Natural Killer/T-Cell Lymphoma (ENKTL): A Mechanism and Prognostic Study

<h3>Purpose/Objective(s)</h3> Extranodal natural killer/T-cell lymphoma, nasal type (ENKTL) is characterized with Epstein-Barr virus (EBV) infection and often presence of B symptoms (including fever, night sweat, and weight loss), but the mechanisms of B symptoms remain unclear. We sought to identify the association of B symptoms with plasma EBV-DNA copy number and cytokine profiles in patients with ENKTL to provide some explanation as well as prognostic analyses. <h3>Materials/Methods</h3> We retrospectively analyzed data from 681 newly diagnosed patients with ENKTL at our institution from April 2007 to December 2019. Plasma EBV-DNA copy levels at presentation were divided into 4 groups: group A (EBV-DNA = 0 copies/ml), group B (0 < EBV-DNA < 10³ copies/ml), group C (10³ ≤ EBV-DNA < 10⁶ copies/ml), and group D (EBV-DNA ≥ 10⁶ copies/ml). In the subgroup of 167 patients with detailed data of cytokines (including IFN, IL-10 and IL-6), patients were divided into three tertiles based on individual cytokines. We explored the association between EBV-DNA levels, cytokines tertiles, and B symptoms via Cochran-Armitage trend test and Goodman-Kruskal Gamma test. Univariate and multivariate analysis for PFS and OS were performed. <h3>Results</h3> The median age of all the 681 patients was 44 years (range 9-83 years). There were 467 males, and 321 patients had B symptoms. The majority of patients were those aged ≤60 years (84.9%), presented with early-stage disease (Ann-Arbor I or II) (79.3%), and with tumor originating from the upper aerodigestive tract (UADT, 84.7%). There were 158 (23.2%), 143 (20.1%), 284 (41.7%), and 96 (14.1%) patients in EBV-DNA group A, B, C, and D, respectively. The Cochran-Armitage trend test demonstrated an upward trend in the incidence of B symptoms from 25.9%, 36.4%, 53.2%, to 80.2%, in EBV-DNA groups with increasing copy number (<i>P</i> < 0.0001). Among 321 patients with B symptoms, 82.6% had a fever with significance. Similarly, the incidence of fever also increased over the four EBV-DNA load groups, which were 15.8%, 31.5%, 45.4%, and 68.8%, respectively (<i>P</i><0.0001). Further analysis of 167 patients with cytokines data demonstrated IFN, IL-10, IL-6 were correlated with EBV-DNA load (<i>P</i> < 0.0001 for all). In addition, the tertiles level of IFN, IL-10 and IL-6 were significantly associated with the presence of B symptoms (<i>P</i> < 0.0001, <i>P</i> = 0.0001, <i>P</i> = 0.0001, respectively) and fever (<i>P</i> = 0.0002, <i>P</i> = 0.0003, <i>P</i> = 0.0005, respectively). In univariate analysis, EBV-DNA load and B symptoms were significant predictor of PFS (<i>P</i> < 0.0001 for both) and OS (<i>P</i> < 0.0001 for both). Multivariate Cox regression analysis showed B symptoms (<i>P</i> = 0.026), primary tumor location (<i>P</i> = 0.019), and Ann-Arbor stage (<i>P</i> = 0.032) as predictive factors of OS. <h3>Conclusion</h3> This exploratory study provided more evidence for a potential association and mechanism, that the massive replication of EBV in vivo could trigger cytokinemia, which may lead to B symptoms and poor prognosis in patients with ENKTL.

999P NPC patients with post-RT EBV clearance show a higher frequency of peripheral CD8 T-cells expressing chemo-attractant receptors at baseline and during radiation therapy

Endemic form of nasopharyngeal carcinoma (NPC) is an EBV-associated head and neck cancer that is highly inflammatory in nature. Most patients present with locoregionally advanced disease which is treated with radiotherapy (RT) with or without chemotherapy. Studies show that plasma EBV DNA level is a prognostic factor that may predict the risk for recurrence. In this study, we have elucidated pre-treatment as well as serial changes of immune markers related to CD8+ T-cell subsets in the blood of NPC patients undergoing RT and investigated whether these changes are associated with plasma clearance of EBV DNA during RT.

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.

Prevalence of Epstein-Barr Viral DNA among children at a single hospital in Suzhou, China

ObjectiveThis study aimed to describe the prevalence of Epstein-Barr virus (EBV)-DNA among children in Suzhou, and to explore the association between plasma EBV load and disease diagnosis. MethodsAll children admitted to the Children's Hospital of Soochow University between January 2018 and September 2020 and subjected to the plasma EBV-DNA assay were included. The authors retrospectively collected demographic and discharge diagnostic information of the participants, and ascribed the disease distribution characteristics of children with positive plasma EBV-DNA by age and viral load. ResultsA total of 38,175 patients underwent plasma EBV-DNA PCR assay, of which 2786 (7.3%) had EBV-DNA in their plasma. Children aged 3–4 years had a high prevalence of EBV infection. Plasma EBV positivity was common with infectious mononucleosis (IM, 40.0%), respiratory infection (20.1%), atypical EBV infection (14.2%), acute leukemia (6.4%), hemophagocytic lymphohistiocytosis (HLH, 4.8%), and idiopathic thrombocytopenic purpura (ITP, 2.9%). With increasing age, plasma EBV positivity was more common in children with IM and atypical EBV infection. However, an inverse correlation was observed in children with respiratory infections and ITP. High levels of EBV loads were more likely to occur in HLH, IM, and atypical EBV infection, especially in HLH. However, lower viral loads were found in respiratory infection and acute leukemia. ConclusionsThis is a large sample study that revealed the prevalence of plasma EBV-DNA levels in children of various ages and presenting illnesses.

Prognostic significance of a combined and controlled nutritional status score and EBV-DNA in patients with advanced nasopharyngeal carcinoma: a long-term follow-up study.

Objective:Several studies have reported that the controlling nutritional status (CONUT) score is a prognostic predictor for survival among patients with different types of cancer. We assessed the prognostic value of changes in the CONUT score during treatment and the ΔCONUT-EBV DNA score in patients with advanced nasopharyngeal carcinoma (NPC).Methods:We retrospectively analyzed 433 patients with advanced NPC having no evidence of metastasis from January 2007 to June 2011; the patients underwent radical concurrent chemoradiotherapy (CCRT) at Sun Yat-sen University Cancer Center and were grouped based on their ΔCONUT and ΔCONUT-EBV DNA scores. Kaplan-Meier curves were used to compare the patient outcomes according to the cut-off ΔCONUT score and the ΔCONUT-EBV DNA scoring system.Results:Among all patients, overall survival (OS) was independently predicted by a high ΔCONUT score (P = 0.031) and high EBV DNA (P < 0.001). The ΔCONUT-EBV DNA score [OS area under the curve (AUC) = 0.621; progression free survival (PFS)-AUC = 0.612; distant metastasis-free survival (DMFS)-AUC = 0.622] was more predictive of OS, PFS, and DMFS in patients with advanced NPC than the ΔCONUT score (OS-AUC = 0.547; PFS-AUC = 0.533; DMFS-AUC = 0.522) and pretreatment plasma EBV DNA levels alone (OS-AUC = 0.600; PFS-AUC = 0.591, DMFS-AUC = 0.610). The ΔCONUT-EBV DNA score was significantly correlated with OS, PFS, and DMFS in patients with advanced NPC treated with CCRT.Conclusions:The ΔCONUT-EBV DNA score may be useful in clinical practice as a convenient biomarker for predicting the outcomes in patients with advanced NPC treated with CCRT.

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.

Adjuvant capecitabine in locoregionally advanced nasopharyngeal carcinoma: A multicenter randomized controlled phase III trial.

6005 Background: We conducted a multicenter, randomized controlled phase III clinical trial (NCT02143388) to investigate the efficacy and toxicity of adjuvant capecitabine (AC) in addition to concurrent cisplatin and radiotherapy (CCRT) compared to CCRT alone in high-risk locoregionally advanced nasopharyngeal carcinoma (LANPC) patients. Methods: Eligibility criteria included AJCC/UICC 7th ed TNM stage III-IVb and one of the following features: T3-4N2 or T1-4N3 or pre-treatment plasma EBV DNA concentration of &gt;20,000 copy/ml or gross primary tumor volume (GTVnx) of &gt;30 cm3 or a maximum standard uptake value (SUVmax) of &gt;10.0 by 18FDG PET-CT within the primary tumor or multiple neck node metastases, with any larger than 4 cm. All patients were randomly assigned in a 1:1 ratio to receive CCRT (3-weekly cisplatin at 100 mg/m2 for 2-3 cycles) followed by AC (1000 mg/m2 bi-daily for 14 days every 21-day cycle for 8 cycles), or CCRT alone. The prescribed radiation doses were 68-72 Gy/30-32 fractions to the PTVnx, 60-68 Gy/30-32 fractions to PTVnd, 60-64Gy/30-32 fractions to PTVhigh-risk, 54-58Gy/30-32 fractions to PTVlow-risk. Primary end point was failure-free survival (FFS). Results: Between Mar 2014 to Jul 2018, 180 patients were recruited (90 patients in CCRT+AC arm and 90 in CCRT alone arm). All patients completed RT and ≥2 cycles of concurrent cisplatin in both treatment arms (cumulative dose intensities for cisplatin were 200 mg/m2 in both arms). 85 (94.4%) patients went on to receive AC, with 71 (78.9%) patients completing 8 cycles; 19 (22.4%) patients had dose reduction of AC. With a median follow-up of 44.8 mo, the 3-y FFS was significantly superior in the CCRT+AC arm than the CCRT arm for the intention-to-treat cohort (87.7% vs 73.3%; HR: 0.52 [95% CI: 0.29-0.77], P = 0.037). 3-year overall, distant metastasis-free and locoregional relapse-free survival were 92.6% vs 88.9% (HR [95% CI]: 0.66 [0.28-1.59]), 88.8% vs. 81.1% (HR: 0.67 [0.33-1.33]) and 91.5% vs 80.0% (HR: 0.50 [0.25-1.00]), respectively. Incidences of G3-4 acute toxicities were 57.8% (52 of 90) in CCRT+AC arm and 51.1% (46 of 90) in CCRT alone arm, with a higher incidence of hand foot syndrome (3.5% vs 0%), xerostomia (11.1% vs 3.3%), mucositis (23.3% vs 16.7%), and anemia (5.6% vs 2.2%) in the CCRT+AC arm. G3-4 late toxicities occurred in 13.3% (12 of 90) and 9.0% (8 of 89), respectively. Conclusions: The addition of capecitabine to CCRT conferred a superior disease control than CCRT alone in high-risk LANPC. Survivals in ITT and PP set. Clinical trial information: NCT02143388. [Table: see text]

Establishment and validation of a prognostic nomogram to predict early metastasis in nasopharyngeal carcinoma patients within six months after radiotherapy and to guide intensive treatment

PurposeThis study aimed to establish an effective prognostic nomogram to predict the risk of early metastasis (EM) in nasopharyngeal carcinoma (NPC) patients, as a guide for intensive treatment. Materials and methodsA total of 9021 patients with biopsy-confirmed NPC at our institute were enrolled in this study between December 2006 to December 2016. We randomized these patients using a proportion of 2/3 and 1/3 and selected 6044 and 2977 patients as the training and validation cohorts, respectively. All patients received radiotherapy with or without chemotherapy. Univariate and multivariate logistical regressions were used to identify independent risk factors. The nomogram’s predictive value was evaluated by concordance indexes (C-indexes), calibration curves, probability density functions (PDFs), and clinical utility curves (CUCs). ROC analysis using Delong test was used to compare efficiency between the nomogram and other risk factors. ResultsIn total, 174 (2.9%) and 81 (2.7%) patients in training and validation cohorts, respectively, had EM.Pretreatment plasma EBV DNA, N stage, LDH, ALP, BMI, and sex were independent predictive factors of EM. The C-indexes of nomogram were 0.756 (95% CI = 0.719–0.793) and 0.766 (95% CI = 0.720–0.813), in the training and validation cohorts, respectively. The C-index of the nomogram was significantly superior to any one of independent factors. According to the PDFs and CUCs and considering the balance of the true positive EM patients and true positive non-EM patients, we chose 5.0% as a threshold probability for clinical decision-making, which could distinguish about 85% and 48% of non-EM and EM patients, respectively. ConclusionOur nomogram had good accuracy in predicting EM incidence, and a 5.0% threshold was appropriate for clinical decision-making.

Characteristics of primary Epstein-Barr virus infection disease spectrum and its reactivation in children, in Suzhou, China.

Epstein-Barr virus (EBV) is a globally prevalent herpesvirus associated with multiple diseases. This study aimed to determine the characteristics of primary EBV infection disease spectrum and reactivation in children, in Suzhou, China. All children admitted to the Children's Hospital of Soochow University between May 2018 and September 2020 with suspected EBV-associated disease and subjected to the indirect immunofluorescence assay for EBV-specific antibodies and plasma EBV-DNA assays were included. Of the 3567 children, 2782 (78.0%) tested positive for EBV. The positive rates of viral capsid antigen (VCA)-IgM, VCA-IgG, early antigen (EA)-IgG, nuclear antigen (EBNA)-IgG, and plasma EBV-DNA were 12.1%, 74.6%, 37.9%, 35.6%, and 31.1%, respectively. The lowest VCA-IgG and EBNA-IgG seropositivity rates occurred at ages between 8 and 36 months, then increased gradually in the older age groups. The EBV-IgM seropositivity rate was the highest in those aged 36 to <72 months. Primary EBV infection was more common in children aged 36 to <72 months. In past infections, reactivation mainly occurred in 8 to <36 months. The most common disease caused by primary EBV infection was infectious mononucleosis (56.1%), followed by a respiratory infection (17.0%). Respiratory infection (30.0%), EBV infection (29.2%), and hemophagocytic lymphohistiocytosis (HLH) (15.8%) were the commonest diseases caused by EBV reactivation. EBV reactivation was a risk factor for HLH (adjusted odds ratio, 16.4; 95% confidence interval, 7.9-34.0). Among reactivated patients, the viral load of HLH was higher than that of EBV infection and respiratory infection (p < .01). This is a retrospective large sample study that explored the characteristics of primary EBV infection disease spectrum and reactivation in children.

The exact Deauville score, NABS score and high SUVmax predicts outcome in extranodal natural killer/T-cell lymphoma.

Natural killer T-cell lymphoma (NKTCL) is an aggressive type of non-Hodgkin's lymphoma. While FDG-PET/CT imaging has been increasingly utilized for disease assessment, its prognostic value and potential utility in NKTCL patient stratification remain controversial. We aim to investigate the prognostic utility of FDG-PET/CT and its role in complementing clinical indices. We conducted a retrospective review of 72 patients from a tertiary National Cancer Centre with biopsy-proven NKTCL and available FDG-PET/CT data (either baseline, end of treatment or both). Survival analysis was performed using the Kaplan-Meier method and multivariable Cox proportional regression. High initial SUVmax was significantly associated with advanced Ann-Arbor stage (p = 0.0352), elevated LDH levels (p = 0.0059) and plasma EBV DNA detection (p = 0.0278). SUVmax correlated with worse progression-free survival (PFS) (HR 3.68, 95% CI 1.56-8.69, p = 0.0030) and a trend toward worse overall survival (OS) (HR 2.06, 95% CI 0.95-4.45, p = 0.0676). End of treatment Deauville scores of 4-5, as compared to scores of 1-3, was associated with worse PFS (HR 2.72, 95% CI 1.04-7.12, p = 0.0419). Notably, while all patients with scores of 5 developed progressive disease, only 2 of 5 patients with scores of 4 eventually relapsed. Clinical indices (NABS score) were still able to stratify survival outcomes regardless of end-of-treatment Deauville scores. A Deauville score of 5 is more diagnostic of true disease progression than a score of 4, and NABS score may be used in patients who achieve Deauville scores of 1-3 for further risk stratification. A higher SUVmax at baseline portends a worse prognosis in NKTCL.

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.

Analysis on Clinical Characteristics and Prognosis of 47 Patients with Extranodal NK/T Cell Lymphoma

To investigate the clinical features and prognostic factors of patients with extranodal NK/T cell lymphoma (ENKTL). The clinical data of patients with ENKTL from November 2009 to November 2019 was collected and retrospectively analyzed to clarify the clinical features of ENKTL, and evaluate the factors that affected survival and prognosis. Forty-seven patients with ENKTL were collected, median age was 40 (12-82) years old, and more common in males than females, at the ratio of 1.47∶ 1. The median follow-up was 28 (1-112) months, and 5-year overall survival (OS) rate was 49.3%. The 5-year OS rates of the subjects with ECOG performance stage 0-1 and ≥2 were 51.6% and 0 (P=0.001), respectively. The 5-year OS rates of International Prognostic Index (IPI) score 0-1 and ≥2 were 60.0% and 40.6% (P=0.027), respectively. The 5-year OS rates of Ann Arbor staging Ⅰ/Ⅱ and stage Ⅲ/Ⅳ were 61.3% and 31.7% (P=0.005), respectively. The 5-year OS rates of the patients with presentation of B symptoms and without presentation of B symptoms were 79.0% and 30.1% (P=0.013), respectively. The 5-year OS rates of plasma EBV-DNA level < 5×102/ml and ≥ 5×102/ml were 60.4% and 33.3% (P=0.003), respectively. The 5-year OS rates of the patients receiving chemotherapy alone, combined chemotherapy and radiotherapy, and chemotherapy followed by autologous hematopoietic stem cell transplantation (auto-HSCT) were 12.9%, 86.5%, and 62.5% (P=0.001), respectively. Univariate analysis showed that ECOG score, IPI score, Ann Arbor staging, B symptoms, the copy number of EBV-DNA, and treatment regimens were statistically significant for OS. Multivariate analysis showed that ECOG score, B symptoms, the copy number of EBV-DNA, and treatment regimens were independent factors of ENKTL OS. ECOG score, B symptoms, the copy number of EBV-DNA, and treatment regimens are independent prognostic factors for OS of patients with ENKTL.

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.

Nomogram for the prediction of primary distant metastasis of nasopharyngeal carcinoma to guide individualized application of FDG PET/CT.

This study aimed to establish an effective nomogram to predict primary distant metastasis (DM) in patients with nasopharyngeal carcinoma (NPC) to guide the application of PET/CT. In total, 3591 patients with pathologically confirmed NPC were consecutively enrolled. The nomogram was constructed based on 1922 patients treated between 2007 and 2014. Multivariate logistical regression was applied to identify the independent risk factors of DM. The predictive value of the nomogram was evaluated using the concordance index (C-index), calibration curve, probability density functions (PDFs), and clinical utility curve (CUC). The results were validated in 1669 patients enrolled from 2015 to 2016. Net reclassification improvement (NRI) was applied to compare performances of the nomogram with other clinical factors. The best cut-off value of the nomogram chosen for clinical application was analyzed. A total of 355 patients showed primary DM among 3591 patients, yielding an incidence rate of 9.9%. Sex, N stage, EBV DNA level, lactate dehydrogenase level, and hemoglobin level were independent predictive factors for primary DM. C-indices in the training and validation cohort were 0.796 (95% CI, 0.76-0.83) and 0.779 (95% CI, 0.74-0.81), respectively. The NRI indices demonstrated that this model had better predictive performance than plasma EBV DNA level and N stage. We advocate for a threshold probability of 3.5% for guiding the application of PET/CT depending on the clinical utility analyses. This nomogram is a useful tool to predict primary DM of NPC and guide the clinical application of PET/CT individually at the initial staging.

Role of Viral Infections in Multiple Sclerosis Pathogenesis among Indian Population.

The role of viral infections in multiple sclerosis (MS) pathogenesis is unclear. Certain neurotropic viruses previously linked with MS among white population were studied including Epstein-Barr virus, human herpesvirus-6 (HHV-6) and MS-associated retrovirus (MSRV). Sixty-two MS patients (37 had a recent clinical relapse) and 65 controls with other neurological disorders were included. Blood and cerebrospinal fluid (CSF) samples were obtained and processed with the primary objective of determining whether there was intrathecal multiplication of viruses under study (EBV, HHV6 A and B and human endogenous retrovirus) or a breach in blood-brain barrier associated with viral presence in both peripheral blood and CSF. Evidence of breach in blood-brain barrier was seen in 86.5% of patients as evidenced by abnormal CSF/serum albumin index and or MRI. EBV nuclear antigen (EBNA1 IgG) was seen in 89% of MS patients and 58% controls (P = <0.001). However, HHV6 IgG was similar in both groups (85% versus 81%; P = 0.45). In affinity immunoblotting reaction intrathecal IgG synthesis against EBNA1 antigen was demonstrable in 26% (16/62) of patients and none against HHV6. A subset of patients showed significant elevation in mean copy number of plasma EBV DNA during relapse and there was a trend for the same among patients harboring HHV-6B. No evidence of isolated intrathecal viral presence or multiplication was seen. The results of our study suggest that viruses studied namely EBV and HHV6 have a role in triggering relapses through a peripheral mechanism, rather than a direct role through intrathecal multiplication.

Superiority of Epstein-Barr Virus DNA in the Plasma Over Whole Blood for Prognostication of Extranodal NK/T Cell Lymphoma.

BackgroundExtranodal natural killer T cell lymphoma (ENKTL) is a rare subtype of non-Hodgkin lymphoma with invariable infection of lymphoma cells with Epstein-Barr virus (EBV), and the presence of EBV-DNA in the blood is a well-known prognosticator. However, there is no consensus on which blood compartment is more optimal for predicting survival outcomes.MethodsWe analyzed 60 patients who were newly diagnosed with ENKTL from a prospectively collected database. EBV-DNA was measured in the whole-blood (WB) and plasma at the time of diagnosis and after treatment completion.ResultsEBV-DNA was detected in pre-treatment WB and plasma in 37 (61.7%) and 23 (38.3%) patients, respectively. The presence of pre-treatment plasma EBV-DNA was significantly associated with advanced stage while presence of WB EBV-DNA did not. Positivity of pre-treatment plasma-EBV, but not WB EBV-DNA, was independently associated with poor PFS (HR, 4.22;95% CI, 1.79–9.97; P=0.001) and OS (HR, 8.38; 95% CI, 3.03–23.19; P<0.001) in the multivariate analysis. After treatment completion, positivity of plasma-EBV was independently associated with poor PFS (HR, 9.41; 95% CI, 2.27–39.02; P=0.002) and OS (HR, 32.38; 95% CI, 3.25–322.56; P=0.003), whereas no significant association was observed between WB-EBV status and survival outcomes.ConclusionsOur results suggest that EBV-DNA in the plasma has better prognostic values than WB in patients with ENKTL.

Diagnostic value of serological and molecular biological tests for infectious mononucleosis by EBV in different age stages and course of the disease.

Epstein-Barr virus (EBV)-based serologic antibody and viral nucleic acid assays have been found to be feasible means to diagnose infectious mononucleosis (IM) caused by EBV in children. In this study, we will further explore theirdiagnostic value for IM by EBV in different age stages and over the course of the disease. A collection of 616 children from clinically suspected IM cases was studied. Indirect immunofluorescence (IIF) for EBV-specific antibody (Euroimmun) combined with plasma EB viral nucleic acid assay (real-time fluorescence quantitative polymerase chain reaction reverse-transcription polymerase chain reaction) were used as reference methods. The diagnostic efficiency of the peripheral blood routine test, serologic antibody test, and plasma EB viral nucleic acid assay for the diagnosis of IM was evaluated, respectively. The sensitivity, specificity, Youden' index and the area under curve (AUC) were 93.08%, 87.77%, 0.81 and 0.904 (95% confidence interval [CI]: 0.878-0.931) for the peripheral lymphocyte test (lymphocytosis > 5 × 109 /L), 98.27%, 91.13%, 0.89 and 0.947 (95% CI: 0.927-0.967) for the plasma EBV-DNA test, and 84.08%, 96.33%, 0.80 and 0.902 (95% CI: 0.874-0.930) for the EBV viral capsid antigen (VCA)-IgG avidity test. The plasma EBV-DNA test has ahigher diagnostic value than the VCA-IgG avidity test in children aged <6 years, especially aged <3 years; the peripheral lymphocyte test and plasma EBV-DNA test are suitable for the early stage of the disease, while the VCA-IgG avidity test for after 7 days of the disease. EBV antibody detection (IIF) should be combined with EBV nucleic acid detection in children age <6 years and the early stage of the disease.