Epstein-Barr Virus Serological Markers Research Articles

Differences in serological profiles of Epstein-Barr virus infection in patients with head and neck tumors and lymphomas.

e18055 Background: Epstein-Barr virus (EBV) is associated with the development of various cancers, including nasopharyngeal carcinoma (NPC), gastric cancer, and lymphomas. The use of EBV serological markers in screening and monitoring of NPC has been shown to be valuable. The significance of serological markers in the diagnosis of other head and neck cancers is poorly described. The aim of this study was to analyze the serological profiles of EBV infection in patients with head and neck tumors. Methods: The main group included 24 patients with laryngeal cancer (n = 12) and oral mucosa cancer (n = 12). Keratinizing squamous cell carcinoma was registered in 22 (91.7%) patients, and adenoid cystic cancer in 2 (8.3%) patients. The control group included 44 lymphoma patients. Antibodies to EBV proteins were determined in the blood serum by ELISA and Western blot (WB) tests. Results: ELISA detected antibodies of the A, M and/or G classes against various EBV proteins in 100% of patients in the main group and 97.7% of controls. IgA VCA in patients with head and neck cancer were determined 2.7 times more often (50% vs 18.2%, p = 0.034) than in patients with lymphomas, IgG EA - 2.1 times more often (58.3% vs 27.3%, p = 0.049), and the complex of acute infection markers (IgA VCA, IgM VCA, IgG EA in various combinations) was determined 2.0 times more often (66.7% vs 34.1%, p = 0.045). Atypical profile (only VCA IgG+) was determined only in patients with lymphomas (4.5%). The profile characteristic of immunosuppression (IgG VCA+, IgG EA+, NA IgG±) was determined 1.8 times more often in patients with head and neck cancer than in patients with lymphomas (25% vs 13.6%, p > 0.05). According to WB tests, patients of the main group more often demonstrated IgM EBNA-1p79 (16.7%), VCA p65 (16.7) and p22 (16.7%), while controls – VCA p22 (27.8%), EA-R p93 (16.7%) and VCA p33 (16.7%). IgG in the main group was more often determined to EBNA-1 p79 (91.7%), VCA p22 (91.7%) and VCA p33 (66.7%), in the control group to VCA p40 (87.5%), EBNA-1 p79 (75%) and VCA p22 (62.5%). Statistically significant differences were found only for VCA p42 (0% in the experimental group and 37.5% in controls, p = 0.049), VCA p40 (25% and 87.5%, p = 0.009) and p27 (0% and 37.5%, p = 0.049). EA-D p45 was determined only in patients with lymphoma. Conclusions: The vast majority of patients in both groups were previously infected with EBV. Serological profiles of EBV infection in patients with head and neck cancer and lymphomas were significantly different. Markers of acute infection were more often determined in patients with head and neck cancer; the range of individual proteins in the main group was narrower than in the control group. The biological meaning of the differences in the detection rates of antibodies to individual EBV proteins in patients of the two groups remains to be elucidated.

Association Between Serum Cotinine Level and Serological Markers of Epstein-Barr Virus in Healthy Subjects in South China Where Nasopharyngeal Carcinoma Is Endemic.

Introduction: Self-reported smoking has been associated with higher seropositivity for the IgA response to Epstein–Barr virus (EBV) viral capsid antigen (VCA-IgA) and transcription activator protein (Zta) in healthy men in southern China where nasopharyngeal carcinoma (NPC) is endemic. Results on the association of biochemically verified smoking status with EBV reactivation are scarce. We aimed to investigate the relations of serum cotinine level with serological markers of EBV in healthy women, in addition to men.Methods: We collected information on demographic, lifestyle, environmental factors, and EBV serological markers in a cross-sectional study on 2,275 healthy subjects who were recruited from physical examination centers in Guangdong Province, China. In the present analysis, 901 subjects' serum cotinine levels have been measured by enzyme-linked immunosorbent assay (ELISA). Odds ratios (seropositivity of four EBV serological markers vs. seronegativity) for categorical serum cotinine levels were calculated by unconditional logistic regression with a group-specific confidence interval (CI).Results: In women, compared with lower serum cotinine level (0–0.71 ng/ml), higher cotinine level (>0.71–1.20 ng/ml; >1.20–228.40 ng/ml) was associated but non-significantly with higher seropositivity for EBV VCA-IgA (age- and education-adjusted OR = 1.18, 95% CIs = 0.84–1.64, 1.06, 0.75–1.50). These associations remained but still non-significant after adjusting for 5-year age group, education, family history of cancer, consumption of tea, Chinese herbal tea, salted fish at childhood, and exposure to occupational dust, chemical, fume, and radiation (multivariable adjusted OR = 1.21, 95% CIs = 0.85–1.71, 1.09, 0.76–1.55). In men, compared with lower serum cotinine level (0–2.15 ng/ml), higher cotinine level (>2.15–103.6 ng/ml; >103.6–419.4 ng/ml) was significantly associated with higher seropositivity for EBV VCA-IgA and Zta-IgA (age- and education-adjusted OR = 2.16, 95% CIs = 1.37–3.41, 1.79, 1.11–2.90; 1.98, 1.17–3.34, 1.95, 1.14–3.34). The association remained significant after adjusting for potential confounders for Zta-IgA (OR = 2.32, 95% CI = 1.37–3.93 for >2.15–103.6, and 2.50, 1.43–4.38 for >103.6–419.4 ng/ml), but not for VCA-IgA (2.06, 1.29–3.27, and 1.61, 0.96–2.71).Conclusions: Higher serum cotinine level is associated with higher seropositivity for EBV serological markers in healthy men in southern China. Such positive association was also observed in women but became non-significant. If confirmed to be causal, this finding has important implications for tobacco control and prevention of EBV-related disease, particularly for NPC.

EPSTEIN-BARR VIRUS AND NASOPHARYNGEAL CARCINOMA: VIRAL MARKERS FOR DIAGNOSTICS AND ASSESSMENT OF CLINICAL STATUS OF PATIENTS.

The etiological role of the Epstein-Barr virus (EBV) in the development of an undifferentiated histological variant of nasopharyngeal carcinoma (uNPC) found for the first time in regions with a high incidence of this pathology, the Southern provinces of China and the countries of Southeast Asia, and later in the rest of the world, has served as a basis for the widespread use of EBV serological markers for the diagnosis of this form of tumor. In recent years, the use of a test based on the quantitative determination of the EBV DNA concentration in the blood plasma of uNPC patients for early detection and monitoring of the disease has become widespread in endemic regions. In non-endemic regions, such studies virtually have not been carried out, and moreover, the comparative evaluation of the significance of two viral markers, serological and EBV DNA load in the bloodstream of uNPC patients, for diagnostics and evaluation of the therapeutic effect was not investigated. The aim of this study was to compare the clinical value of two serological markers and plasma EBV DNA load in uNPC patients from non-endemic region (Russia). The obtained results indicate that IgA antibodies to the viral capsid antigen (IgA/VCA) and plasma EBV DNA concentration can be successfully used for the diagnosis of uNPC, while IgG/VCA antibodies have no practical significance as an uNPC marker. In addition, it was found that plasma EBV DNA load is more sensitive marker of uNPC than IgA/VCA titers because DNA copy numbers reflect more accurately the effect of the therapy and the clinical state of patients at the stages of remission or relapse. It was shown for the first time that in the non-endemic region the simultaneous evaluation of IgA/VCA antibody levels and the plasma EBV DNA loads are the most effective markers for the diagnostics of uNPC. However, we believe, that it is more practical to use IgA/VCA antibody levels for uNPC screening, and plasma EBV DNA copies - for monitoring of the disease.

Comparison of two automated instruments for Epstein-Barr virus serology in a large adult hospital and implementation of an Epstein-Barr virus nuclear antigen-based testing algorithm.

Serology remains the mainstay for diagnosis of Epstein-Barr virus (EBV) infection. This study compared two automated platforms (BioPlex 2200 and Architect i2000SR) to test three EBV serological markers: viral capsid antigen (VCA) immunoglobulins of class M (IgM), VCA immunoglobulins of class G (IgG) and EBV nuclear antigen-1 (EBNA-1) IgG. Using sera from 65 patients at various stages of EBV disease, BioPlex demonstrated near-perfect agreement for all EBV markers compared to a consensus reference. The agreement for Architect was near-perfect for VCA IgG and EBNA-1 IgG, and substantial for VCA IgM despite five equivocal results. Since the majority of testing in our hospital was from adults with EBNA-1 IgG positive results, post-implementation analysis of an EBNA-based algorithm showed advantages over parallel testing of the three serologic markers. This small verification demonstrated that both automated systems for EBV serology had good performance for all EBV markers, and an EBNA-based testing algorithm is ideal for an adult hospital.

Epstein-Barr virus biomarkers for nasopharyngeal carcinoma in non-endemic regions.

The Epstein-Barr virus (EBV) plays a key role in the development of undifferentiated nasopharyngeal carcinoma (uNPC). In uNPC endemic regions EBV-specific antibodies and plasma EBV DNA load are used as markers for the early detection of uNPC and monitoring of the disease. In non-endemic regions, such studies were practically not conducted. The aim of this study was to compare the clinical significance of EBV serological markers and plasma EBV DNA levels for uNPC patients in a non-endemic region, Russia. The results obtained indicate that both viral capsid antigen/immunoglobulin A (VCA/IgA) antibodies and plasma EBV DNA copies can effectively be used for nasopharyngeal carcinoma (NPC) diagnosis. Besides, plasma EBV DNA load was found to be a more sensitive marker of uNPC than VCA/IgA antibody titres, as it reflected the effect of the therapy in stages of remission and relapse of the disease more precisely. Our study, for the first time, demonstrates that the simultaneous use of plasma EBV DNA loads and VCA/IgA antibody levels are indispensable markers for uNPC in non-endemic regions: a serological marker can be more effectively used for NPC screening, but EBV DNA copies are better for monitoring the disease. However, both markers turned out to be practically unsuitable for assessing the clinical status of patients. Serological markers did not correlate with any signs of the tumour process estimated by tumour, node and metastasis (TNM) classification and the plasma EBV DNA loads correlated only with the size of the pathologically altered lymph nodes (N). Additional study is required to confirm these findings.

Diagnostic value of the Epstein-Barr virus serological markers in patients with nasopharyngeal carcinoma in cases of undetectable primary tumor location

The goal of this work was to describe a method for diagnosis of the non-keratinizing nasopharyngeal carcinoma (nNPC) in cases of the undetectable primary tumor location. The method is based on evaluation of IgG and IgA antibody levels to the capsid (VCA) and early antigens (EA) of the Epstein-Barr virus (EBV). The diagnosis of nNPC is established by a so-called decision rule. The latter was created by mathematical processing of the method of multifactor analysis of the results of anti-EBV antibody testing of 72 patients with clinically and morphologically confirmed nNPC and 72 patients with other head and neck benign tumors (OHNT) not associated with EBV, which were tested as a control group. The diagnostic value of the decision rule which was tested in the group of 77 patients with confirmed nNPC and 231 patients of a control group was high. The numbers of false negative and false positive cases were equal to 5.2% (4/77) and 6.5% (17/231), respectively. Among 32 patients with undetectable primary tumors the decision rule was able to identify 11 cases of nNPC. This diagnosis later was confirmed by morphological and instrumental methods of study. Only in two cases, false negative result was obtained (2/32; 6.3%) indicating that the serological diagnostics of nNPC with the decision rule is highly specific but not exact. Thus, the data obtained allowed us to conclude that the serological testing of EBV specific antibody evaluated by the decision rule can be recommended as an important test supplementing the standard methods of pdNPC diagnostics including cases with undetected primary tumor location.

Investigation of Epstein-Barr Virus and Parvovirus B19 DNA in Allogeneic Stem Cell Transplant Patients

Objective: We aimed to investigate posttransplant Epstein-Barr virus (EBV) and parvovirus B19 DNA in allogeneic stem cell transplant patients between 2009 and 2010. Materials and Methods: Forty-five adult patients in whom allogeneic stem cell transplantation was performed between April 2009 and November 2010 in the Erciyes University Faculty of Medicine, Department of Internal Medicine, Division of Hematology and Oncology, were included in the study. EBV and parvovirus B19 DNA positivity was investigated by using real-time polymerase chain reaction technique in 135 plasma samples obtained after transplantation at between 1 and 6 months. Pretransplant serological markers of EBV and parvovirus B19 were provided from patient files. Results: In 32 (71.1%) of the patients, EBV antibodies in the pretransplantation period were as follows: anti-EBNA-1 IgG (+), VCA IgM (-), and VCA IgG (+). In 2 patients (4.45%), these antibodies were as follows: anti-EBNA-1 IgG (+), VCA IgM (-), and VCA IgG (-). In 1 patient (2.2%), they were as follows: anti-EBNA-1 IgG (-), VCA IgM (-), and VCA IgG (+). EBV serological markers were negative in 2 (2.2%) out of 45 patients before transplantation. There was low DNA positivity (<600 copies/mL) in 4 patients (8.9%), and VCA IgM was negative and VCA IgG was positive in these same 4 patients. In spite of low viral load, there were no symptoms related to EBV, and posttransplant lymphoproliferative disorder (PTLD) did not occur. While in 44 (99.7%) of 45 patients parvovirus B19 IgM was negative and IgG was positive, parvovirus B19 IgM was positive and IgG was negative in 1 (2.3%) patient. Parvovirus B19 DNA was not identified in any of the samples obtained from these 45 patients. Conclusion: In this study, EBV and parvovirus B19 DNA were investigated in allogeneic stem cell transplant patients. None of the patients developed PTLD and parvovirus B19 DNA positivity was not detected. However, this issue needs to be further evaluated in prospective, multicenter studies with larger series of patients.

Correlates of anti-EBV EBNA1 IgA positivity among unaffected relatives from nasopharyngeal carcinoma multiplex families

Background:To determine whether non-viral nasopharyngeal carcinoma (NPC) risk factors might be associated with (and mediated through) Epstein–Barr virus (EBV) serological responses linked to NPC risk, we evaluated predictors of risk of anti-EBNA1 IgA seropositivity and other markers among unaffected relatives from a large NPC family study in Taiwan.Methods:Multivariate logistic regression conditioned on family was used to examine the associations between sociodemographic, dietary, lifestyle, and occupational variables and risk of anti-EBV EBNA1 IgA positivity, anti-VCA IgA, and anti-DNase positivity.Results:Among 2393 unaffected relatives from 319 multiplex families, 1180 (49.3%) were anti-EBV EBNA1 IgA seropositive. None of the associations with anti-EBNA1 IgA were statistically significant, except for being 31–50 years of age (vs <30, adjusted ORs 0.51–0.57). For one or more EBV serological markers, there were suggestive associations for older age, GuangDong firm salted fish, betel use, current alcohol use, and male gender.Conclusion:Overall, we found little evidence to suggest that non-viral NPC risk factors significantly alter EBV serological patterns, suggesting that non-viral NPC risk factors act through pathways independent of EBV serological responses.

Antibodies against Epstein–Barr virus gp78 antigen: a novel marker for serological diagnosis of nasopharyngeal carcinoma detected by xMAP technology

Immunoglobulin (Ig) A and/or IgG reactivities to several Epstein-Barr virus (EBV) antigens have been used to facilitate diagnosis of nasopharyngeal carcinoma (NPC). However, antibodies against gp78, an EBV membrane glycoprotein, have not been examined to this day. In this study, we utilized Luminex multi-analyte profiling (xMAP) technology to analyse antibody responses to a synthetic peptide of gp78 in sera samples from 95 NPC patients and 91 healthy controls. Our results showed the sensitivity and specificity of IgA-gp78 for NPC diagnosis were 79 and 71 %, respectively, while those of IgG-gp78 were 74 and 73 %, respectively. The IgA and IgG responses to different EBV antigens were not identical within an individual and IgA-gp78 and IgG-gp78 could be complementary to antibodies against viral capsid antigen (VCA), the diffused early antigen (EA-D) and the nuclear antigen EBNA1 for NPC diagnosis. When the six EBV parameters for NPC prediction, i.e. IgA-gp78, IgG-gp78, IgA-VCA, IgA-EBNA1, IgA-EA-D and IgG-EA-D, are combined, the combined predictors were able to reach overall sensitivity and specificity of 91 and 95 %, respectively. Thus, simultaneous detection of these EBV serological markers could improve the predictive values of NPC using xMAP technology.

The Epstein-Barr Virus and Chronic Fatigue Syndrome

To the Editor.— Based on eight years of clinical experience and having performed many thousands of Epstein-Barr virus (EBV) serological assays with emphasis on IgG anti-early antigen I cannot agree with the conclusion of Hellinger et al1that the antibody to EBV early antigen is not helpful in the clinical evaluation of patients with chronic fatigue syndrome. The initial suspicion of a direct etiologic link between EBV and chronic fatigue syndrome has yielded to the probability that EBV reactivation is a secondary phenomenon related to a primary, transient immune dysfunction, the cause of which is currently unknown. In contrast to all other EBV serological markers, which reflect only previous infection, anti-early antigen does rise and fall with clinical activity of the signs and symptoms used to define the chronic fatigue syndrome.2Although not diagnostic, it is a useful therapeutic marker when initially found in significant elevation (ie, ≥1:80).