Epstein-Barr Virus Capsid Antigen Research Articles

Can the Epstein-Barr Virus Play a Role in the Development of Prostate Cancer?

Prostate cancer (PCa) is the fourth most frequently diagnosed cancer worldwide, accounting for 7.3% of all cancers. PCa mortality is the fifth most common cause of cancer death. Despite well-known factors influencing the development of PCa, such as age, race/ethnicity and family history, many researchers have raised the possibility of persistent infections with oncogenic viruses. Therefore, we aimed to assess the frequency of Epstein-Barr virus (EBV) DNA in tissue collected from PCa patients. Next, the frequency and the level of Epstein-Barr virus capsid antigen (EBVCA) and Epstein-Barr nuclear antigen 1 (EBNA1) antibodies in both IgA and IgG classes were measured. The antibody titer was also analyzed depending on the risk group, Gleason score (GS) and tumor, node, metastasis (TNM) classification. Serum samples were analyzed using the Microblot-Array EBV IgM, IgA and IgG test kits. The study group consisted of 115 patients diagnosed and histopathologically confirmed with PCa. In 49% of patients included in the study, EBV DNA was detected in the tumor tissue. The studies showed both higher seroprevalence and higher antibody titers in patients with EBV-positive PCa compared to patients with EBV-negative PCa. We also observed a dependence of antibody titer on pathological features, such as GS, risk group and T stage.

Serological profile of IGG antibodies to viral capsid antigen of epstein-barr virus in oral pre-cancer and cancer patients

Cancer of the oral cavity is a major and increasing public health problem. Oral cancer is the sixth most common cancer reported globally with an annual incidence of over 300,000 cases, of which 62% arise in developing countries. In India, because of cultural, ethnic, and geographic factors and the popularity of addictive habits, the frequency of oral cancer is high. Despite the various etiological and predisposing factors associated with oral pre-cancer and cancer, the role of co-carcinogenic effects of viruses like HSV has been documented. With this background and considering the high prevalence of oral cancer in India an attempt has been made to study the association of EBV with oral cancer using serological methods.1. To study the IgG antibodies to Viral Capsid Antigen (VCA) of Epstein-Barr virus in oral pre-cancer and cancer by a laboratory standardized Indirect Immuno Fluorescent (IIF) test. 2. To compare and corelate the results of IIF test with a commercially available ELISA for VCA IgG antibodies to EBV. 3. To assess the severity of disease progression in oral pre-cancer and cancer using the serological profile.In the present study, 84 subjects were assessed for serum IgG antibodies to VCA of EBV using the IIF technique and later the results were compared with an ELISA kit. Group I were controls, Group II to Group V were oral squamous cell carcinoma subjects according to the clinical staging. Group VI comprised of oral pre-cancer subjects. Age range was between 40-70 years, and the mean age was 50.31 +/- 8.23 years. All the 42 control sera were positive for IgG antibodies to VCA of EBV but with a significantly low geometric mean titre(GMT) of 8.561.The GMTs of oral squamous carcinoma and pre-cancer sera was positive but progressively increased when compared to control sera and were highly significant (P<0.001).As there was a significant increase in the GMTs in oral squamous cell carcinoma subjects and pre-cancer subjects, serology of EBV may be interpreted to be useful in assessing the severity of the disease and the techniques of IIF and ELISA may be used as first line screeners when large number of subjects need to be assessed.

Humoral response to Epstein-Barr virus in patients with multiple sclerosis treated with B cell depletion therapy.

B cell depletion therapy is highly effective in relapsing-remitting multiple sclerosis (RRMS). However, the precise underlying mechanisms of action for its biological effects in MS have still not been clarified. Epstein-Barr virus (EBV) is a known risk factor for MS and seems to be a prerequisite for disease development. EBV resides latently in the memory B cells, and may not only increase the risk of developing MS, but also contribute to disease activity and disability progression. Therefore, the effects of B cell depletion in MS could be associated with the depletion of EBV-infected cells and the altered immune response to the virus. In this study, we investigate the impact of B cell depletion on the humoral immune response specific to EBV in patients with MS. Newly diagnosed, treatment-naïve patients with RRMS were followed up to 18 months after initiation of B-cell depletion therapy in the Overlord-MS study, a phase III trial (NCT04578639). We analyzed serum sampled before treatment and after 3, 6, 12 and 18 months for immunoglobulin γ (IgG) against Epstein-Barr nuclear antigen 1 (EBNA1) and Epstein-Barr viral capsid antigen (VCA). We analyzed antibodies to cytomegalovirus (CMV) and total IgG in serum, as controls for viral and overall humoral immunity. The risk allele, HLA-DRB1*15:01, and the protective allele, HLA-A*02:01, were determined in all participants. In addition, polymerase chain reaction (PCR) for circulating EBV-DNA was performed in the first 156 samples drawn. The associations between time on B cell-depletion therapy and serum anti-EBV antibody levels were estimated using linear mixed-effects models. A total of 290 serum samples from 99 patients were available for analysis. After 6, 12 and 18 months, the EBNA1 IgG levels decreased by 12.7% (95% CI -18.8 to -6.60, p<0.001), 12.1% (95% CI -19.8 to -3.7, p=0.006) and 14.6% (95% CI to -25.3 to -2.4, p=0.02) respectively, compared to baseline level. Carriers of the HLA-DRB1*15:01 allele had higher EBNA1 IgG levels at baseline (p=0.02). The VCA IgG levels significantly increased by 13.7% (95% CI 9.4 to 18.1, p<0.001) after 3 months, compared to baseline, and persisted at this level throughout the follow-up. CMV IgG levels decreased, but to a lesser extent than the decrease of EBNA1 IgG, and total IgG levels decreased during therapy. Circulating EBV-DNA was found in only three of 156 samples from 64 patients. EBNA1 IgG levels decreased, while VCA IgG levels increased, during B cell depletion therapy. This supports the hypothesis that the mechanism of action for B cell depletion therapy might be mediated by effects on EBV infection, which, in turn, mitigate immune cross-reactivity and disease perpetuation.

High Epstein-Barr virus capsid antigen IgG level associates with the carriership of CD8+ T cell somatic mutations in the STAT3 SH2 domain

High carrier prevalence of STAT3 SH2 domain somatic mutations was recently discovered in CD8+ T cells. We found these low-allele-fraction clones in 26% of donors, without difference between multiple sclerosis (MS) patients and controls. Here we tested whether anti-viral antibodies associate with the carriership of these mutant clones. We compared antibody responses against common viruses in mutation carriers vs. non-carriers. Plasma samples of 152 donors (92 MS patients, 60 controls) were analyzed for antibodies against cytomegalovirus (CMV), Epstein-Barr virus (EBV), human herpesvirus-6A and parvovirus B19. The mutation carrier status associated with EBV VCA IgG level (p = 0.005) and remained significant after logistic regression (p = 0.036). This association was contributed similarly by MS patients and controls. These results suggest that EBV contributes to the generation or growth of these clones. The pathogenic role of the STAT3 mutant clones in MS is presently unclear, but their detailed characterization warrants further study.

Significance of Dynamic Changes of VCA-IgA Levels in Pre- and Post-treatment Plasma of Patients with Nasopharyngeal Carcinoma: Development of a Clinically-Oriented Model.

Nasopharyngeal carcinoma (NPC) responds well to radiotherapy but recurrence and metastasis are common. Currently, there is no widely used biomarker for accurately predicting the recurrence and metastasis of NPC. In this study, we aimed to evaluate the prognostic ability of Epstein-Barr virus (EBV) capsid antigen (VCA-IgA) kinetics by assessing the dynamic changes of VCA-IgA levels in the pre- and post-treatment plasma of patients with NPC and have proposed a prognostic model for clinical use. The clinical records of patients with NPC diagnosed at Sun Yat-sen University Cancer Center were retrieved and classified into a respondent (n=83) or non-respondent (n=25) cohort based on their response to antitumor therapy. Factors associated with the outcomes of the patients were assessed and incorporated in a nomogram. For internal validation, bootstrapping with 1000 resamples was used. The prediction accuracy and discriminative ability of the nomogram were investigated by calibration and concordance index (C-index) and plotted decision curves to assess the benefits of nomogram-assisted decisions in a clinical context. Plasma VCA-IgA level of the non-respondent cohort at the 6th month after treatment was found significantly higher than the respondent cohort. Post-treatment VCA-IgA level, smoking, and distant metastases were identified as independent risk factors for disease-free survival (DFS), and were used to stratify patients with NPC into three risk groups. The median DFS of the low-, middle- and high-risk groups were 48.5, 35.0, and 15.5months, respectively. The C-index of the nomogram was 0.848 (95% CI 0.769-0.926), demonstrating good clinical accuracy for predicting the DFS of patients with NPC. The decision curve showed that the nomogram in predicting DFS was better than VCA-IgA level, smoking, and distant metastases. The proposed VCA-IgA-based nomogram demonstrated a promising ability to predict the DFS of patients with NPC after antitumor therapy. It could be used as a clinical guidance to improve the therapeutic/surveillance strategies of these patients.

Kidney involvement of chronic active Epstein-Barr virus infection.

A 54-year-old Japanese woman presented with a fever of unknown origin and renal dysfunction. She had experienced intermittent fever for 10 years, but the frequency increased to a constant fever. Her renal function had gradually deteriorated over the past year, and serum creatinine and urinary α1-microglobulin levels were elevated to 2.23 mg/dL and 113 mg/L, respectively. Physical and laboratory examinations were unremarkable, except for mild anemia and elevated serum creatinine levels. Analysis of peripheral blood lymphocyte subsets showed 59% CD3, 35% CD4 and 31% CD8. Epstein-Barr virus (EBV) serology showed elevated levels of EBV capsid antigen IgG and EBV nuclear antigen (1:1280 and 1:40, respectively). Plasma EBV-DNA level was 5.63 log IU/mL, and an oligoclonal band was detected by Southern blot hybridization of EBV terminal repeat analysis. Quantification of EBV-DNA in peripheral lymphocyte subset using magnetic beads showed that EBV predominantly resided in CD4+ T-cells. (CD4, 258,768; CD8, 27,738; and CD56, 23,369; IU/µgDNA, respectively). T-cell receptor α gene rearrangement was undetected. Based on the above, she was diagnosed with chronic active EBV infection (CAEBV). 18F-fluorodeoxyglucose positron emission tomography/computerized tomography showed bilateral kidney enlargement and mildly enhanced uptake (maximum standardized uptake value 4.08 g/mL) (Figure 1 top left). Kidney biopsy specimen stained with hematoxylin and eosin showed diffuse tubulointerstitial infiltration of small to medium-sized lymphocytes in the renal parenchyma (Figure 1 top right). Massive infiltration of CD4+ T-cells was documented by immunohistochemical staining (Figure 1 bottom left). Furthermore, EBV-infected cells were detected by in situ hybridization with EBV-encoded RNA (EBER) (Figure 1 bottom right). Immunostaining of EBV nuclear antigen-2 and latent membrane protein 1 were negative. EBER-positive cells were distributed in the same region as CD4+ T-cells, consistent with the predominant presence of EBV-DNA in CD4+ T-cells. Her renal function had improved after initiating chemotherapy, and she is scheduled for hematopoietic stem cell transplantation. The clinical features of EBV infection vary according to the type of EBV-infected cells. The predominant EBV-infected cells in EBV-induced hemophagocytic lymphohistiocytosis are CD8+ T cells, whereas in CAEBV, CD4+ T cells are predominantly infected. Although rare, renal invasion of EBV should be considered in cases with renal impairment, and its clinical significance needs to be investigated. AF and SS wrote and edited the manuscript, provided patient care and reviewed the literature. KS and MT provided patient care. TS and KM wrote and edited the manuscript. All authors reviewed and approved the final version of the manuscript We thank Dr. Yuka Torii (Department of Pediatrics, Nagoya University, Japan) for quantification of the EBV viral load of lymphocyte subsets using magnetic beads. We also thank Dr. Hiroshi Kimura (Department of Virology, Nagoya University, Japan) for his helpful advice on patient care. We appreciate the staff of the Department of Hematology, Kameda Medical Center for providing patient care. We thank Editage (www.editage.jp) for English language editing. The authors declare they have no conflicts of interest. The authors received no specific funding for this work. This study was conducted according to the Declaration of Helsinki, and written consent was obtained from the patient for this report. The patient provided written informed consent for the publication of this case, with the removal of all identifying information to ensure anonymity and maintain privacy. The data that support the findings of this study are available from the corresponding author upon reasonable request. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Epstein-Barr Virus Infection as Significant Predisposing Factor and Post-Radiotherapy Prognostic Indicator among Libyan Patients with Nasopharyngeal Cancer

Background and Objectives: Libya is one of the North African countries which constitute an area of nasopharyngeal carcinoma endemicty, particularly at the north part of Libya as compared to its south portion. Although from the clinical as well as histopathological presentation, there is no uniquely specific pattern of appearance, but there is high deficiency in the data that confirm the possible predisposing factors which may play a role in the development of this common variety of head and neck cancer at this country, and whether these factors affecting the patients response to treatment. Therefore this study was conducted to elucidate the suggestion of the role of Epstein-Barr Virus (EBV) in the risk of nasopharyngeal cancer at Libyan population; the results were correlated with the radiotherapy response and the improvement in the post-therapy prognostic value. Patients and Methods: Sixteen patients aged 9-80 years presented at ENT department -Althowra central teaching hospital-elbyda-Libya at period from September- 2005 up to January-2014 with variable patterns of clinical presentation suggestive nasopharyngeal carcinoma. All patients were evaluated radio- logically as well as endoscopically, the diagnosis was confirmed by the biopsy and further histo-pathological assessment. For all patients the serological elucidation for IgG and IgM anti-EBV antibodies was done, and the results were correlated with the age of the incidence, sex of the incidence, the pattern of clinical presentation, the pattern of histo-pathological presentation, the response to the radiotherapy, the rate of the recurrence, and five years survival. Results: 88% of the patients showed significant increase of serum IgG against Viral Capsid Antigen (VCA) of EBV. 83% of the cases presented with cervical masses and unilateral otitis media with effusion. 86% of cases showed lympho-epithelioma as histo-pathological pattern. On the other hand among all patients, those patients with lympho-epithelioma showed significant response to the concomitant radio- chemotherapy (100%) with high survival rate exceed five years. Conclusion: The EBV infection can be considered as one of main predisposing factors of nasopharyngeal carcinoma at Libyan population. As it was noted from this presenting study that the induction of the cancer by EBV is mainly due to chronic infection rather than acute infection and this was confirmed by significant elevation of serum IgG rather than IgM. In addition it was postulated that the EBV infection most likely to be associated with lympho-epithelioma variety of histo-pathology rather than well-differentiated type, and as it was elucidated via this study that the association of the cancer with EBV infection increased the sensitivity of the tumor to concomitant radio-chemotherpy, thus this will improve the five years survival after the treatment.

Detection of Epstein-Barr virus in systemic sclerosis patients: A molecular and serological based study.

This study aimed to evaluate an association between Epstein-Barr virus (EBV) and systemic sclerosis (SSc). One hundred and fifty (138 female, 12 male) consecutive adult SSc patients fulfilling the American College of Rheumatology (ACR)/ European League Against Rheumatism (EULAR) criteria were included in this cross-sectional study. Serological analysis by line blot for class immunoglobulin G (IgG) and IgM antibodies against EBV antigen (EBV capsid antigen [VCA] gp125, VCA p19, EBNA-1, p22, EA-D) and quantification of EBV DNA in whole blood by real-time polymerase chain reaction was performed. Class IgM antibodies against VCA gp125 (22.8% vs 0%, P < .0002), VCA p19 (55.7% vs 4.4%, P < .0001), EBNA1 (35.7% vs 0%, P < .0001), p22 (24.2% vs 0%, P < .0001), EA-D (14.2% vs 2.2%, P < .04), and class IgG antibodies against p22 (95.7% vs 82.2%, P < .02) and EA-D (54.2% vs 0%, P < .0001) reactivities were significantly higher in SSc patients than in controls. The past infection was significantly associated with the control group (42.8% vs 91%, P < .0001); and the viral reactivation was significantly associated with the SSc group (55.7% vs 4.4%, P < .0001). Only three (2%) out of 150 SSc patients were positive for EBV DNA, similar to the control group (2%) (P > .9). The study shows a strong serological association of EBV (reactivation stage) with SSc patients in the absence of viral DNA in the circulation, indicating the EBV reservoir or tropism presence elsewhere.

Frequent intrathecal production of antibodies to the viral capsid antigen of Epstein-Barr virus in patients with central nervous system post-transplant lymphoproliferative disorder

Primary central nervous system post-transplant lymphoproliferative disorder (PCNS-PTLD) is strongly associated with Epstein-Barr virus (EBV). We analysed intrathecal production of EBV viral capsid antigen (VCA) immunoglobulin (Ig)G and IgA and Epstein-Barr nuclear antigen-1 (EBNA-1) IgG in nine patients with PNCS-PTLD and 20 patients with non-inflammatory neurological diseases (NINDs). Intrathecally produced VCA-IgG was detected in 7/9 (78%), VCA-IgA in 6/9 (67%) and EBNA-1-IgG in 2/9 (22%) patients with PCNS-PTLD, but not in NINDs. This exploratory study suggests that intrathecal EBV antibody production might be frequent in PCNS-PTLD. Detecting intrathecally produced VCA-IgG and VCA-IgA could thus potentially be helpful for diagnosing PCNS-PTLD.

Antibody response to herpes simplex virus-1 is increased in autoimmune encephalitis.

Antibody response to herpes simplex virus-1 is increased in autoimmune encephalitis.

Application of magnetic immunofluorescence assay based on microfluidic technology to detection of Epstein-Barr virus

EB病毒(Epstein-Barr virus, EBV)的早期诊断能够降低患者发生重大疾病的风险。临床上常用的EBV抗体的检测方法存在耗时长、试剂消耗大和效率低等缺点。相比于传统的检测方法,微流控(microfluidics)技术具有高通量、试剂消耗少,污染少和自动化程度高等优点,磁免疫荧光技术具有检测效率高、信号强等优点,将两者的优势结合,能够弥补传统方法的不足。鉴于此,采用聚甲基丙烯酸甲酯(PMMA)作为芯片原材料,经过激光切割及真空热压加工工艺能够快速获得芯片。将包被抗原的磁珠及包被抗人抗体的荧光微球经过冷冻干燥工艺快速冻干成小球并嵌入芯片内,经过孵育和清洗后,进行检测。通过图像分析快速得到检测结果。通过精密度、特异性、剂量-反应曲线及检出限测试,进行性能验证。通过与化学发光免疫分析法(CLIA)检测的临床样本比对,进行方法学与临床应用评价。结果显示相对标准偏差(RSD)均小于10%。与多种常见的病原体抗体均无交叉反应。EB病毒衣壳抗原(Epstein-Barr viral capsid antigen, EB VCA)IgG项目的检出限为1.92 U/mL,线性范围为1.92~200 U/mL,阳性符合率为95.77%(68/71),阴性符合率为86%(43/50); EB VCA IgA项目的检出限为2.79 U/mL,线性范围为2.79~200 U/mL,阳性符合率为92%(46/50),阴性符合率为92.96%(66/71); EB病毒核心抗原1(Epstein-Barr nuclear antigen 1, EB NA1)IgG项目的检出限为3.13 U/mL,线性范围为3.13~200 U/mL,阳性符合率为92.96%(66/71),阴性符合率为92%(46/50); EB NA1 IgA项目的检出限为1.53 U/mL,线性范围为1.53~200 U/mL,阳性符合率为90%(45/50),阴性符合率为91.55%(65/71)。4个项目能在20 min内快速完成检测,且与临床上使用CLIA方法测试的结果具有良好的相关性,可以为临床提供一种快速、灵敏、简便、自动化程度高和易于基层推广的检测方法。

Cytotoxic CD4+ T-cells specific for EBV capsid antigen BORF1 are maintained in long-term latently infected healthy donors.

Epstein Barr Virus (EBV) infects more than 95% of the population whereupon it establishes a latent infection of B-cells that persists for life under immune control. Primary EBV infection can cause infectious mononucleosis (IM) and long-term viral carriage is associated with several malignancies and certain autoimmune diseases. Current efforts developing EBV prophylactic vaccination have focussed on neutralising antibodies. An alternative strategy, that could enhance the efficacy of such vaccines or be used alone, is to generate T-cell responses capable of recognising and eliminating newly EBV-infected cells before the virus initiates its growth transformation program. T-cell responses against the EBV structural proteins, brought into the newly infected cell by the incoming virion, are prime candidates for such responses. Here we show the structural EBV capsid proteins BcLF1, BDLF1 and BORF1 are frequent targets of T-cell responses in EBV infected people, identify new CD8+ and CD4+ T-cell epitopes and map their HLA restricting alleles. Using T-cell clones we demonstrate that CD4+ but not CD8+ T-cell clones specific for the capsid proteins can recognise newly EBV-infected B-cells and control B-cell outgrowth via cytotoxicity. Using MHC-II tetramers we show a CD4+ T-cell response to an epitope within the BORF1 capsid protein epitope is present during acute EBV infection and in long-term viral carriage. In common with other EBV-specific CD4+ T-cell responses the BORF1-specific CD4+ T-cells in IM patients expressed perforin and granzyme-B. Unexpectedly, perforin and granzyme-B expression was sustained over time even when the donor had entered the long-term infected state. These data further our understanding of EBV structural proteins as targets of T-cell responses and how CD4+ T-cell responses to EBV change from acute disease into convalescence. They also identify new targets for prophylactic EBV vaccine development.

Markers of Epstein–Barr virus in clinical assessment of Russian patients with nasopharyngeal cancer

Introduction. Epstein–Barr virus (EBV) is equally widespread in the endemic and non-endemic world regions for nasopharyngeal cancer (NPC). High incidence of NPC in endemic countries and low in non-endemic countries suggest there are different mechanisms and conditions for tumor occurrence and, possibly, different clinical significance of EBV-associated markers. However, significance of these markers for determining NPC in non-endemic regions is still poorly understood. Objective – to determine clinical significance of titers of IgG/IgA antibodies to EBV capsid antigen and concentrations of the viral DNA in patients’ blood plasma as diagnostic and monitoring markers for NPC in a non-endemic region of Russia. Materials and methods. Titers of EB-specific antibodies were determined by indirect immunofluorescence, and concentration of the viral DNA in plasma was measured using a quantitative polymerase chain reaction in real time. Study group included patients with NPC (n = 96), and control group – blood donors (n = 171) and patients with other head and neck tumors (n = 33).Results. Titers of IgG/IgA antibodies to EBV capsid antigen, being an important diagnostic marker of nasopharyngeal cancer, did not always correlate with patients’ clinical condition. Humoral response to emerging events often delayed due to inertia of the immune system. Concentration of EBV DNA in patients’ blood plasma clearly reflected the dynamics of the pathological process: it decreased to background values in remission and increased while the disease progressed. In contrast to endemic regions, we did not find any correlation between the studied EBV markers and clinical manifestations of the disease, evaluated in accordance with the TNM classification (Tumor, Nodus and Metastasis).Conclusion. In non-endemic countries, such as Russia, serological and molecular markers of EBV can be successfully used for the primary diagnosis of NPC. However, for the disease monitoring, it is preferable to use the value of the concentrations of circulating EBV DNA, which, in contrast to the values of IgG/IgA antibody titers to VCA EBV, more accurately reflect the patient’s clinical condition.

Association Between Traditional Herbal Diet and Nasopharyngeal Carcinoma Risk: A Prospective Cohort Study in Southern China.

IntroductionProspective evidence for herbal diet and nasopharyngeal carcinoma (NPC) development is absent. We therefore evaluated the associations of herbal soup and herbal tea with NPC in a prospective cohort study in southern China.MethodsBased on an NPC screening cohort established in 2008–2015, information on herbal diet consumption, potential confounding factors, and Epstein-Barr virus (EBV) antibody levels were collected from 10,179 individuals aged 30–69 years in Sihui city, southern China. Cox regression models were performed to examine herbal diet with NPC risk, and logistic regression models were used to examine herbal diet with EBV reactivation.ResultsDuring a median of 7.54 years of follow-up, 69 participants developed NPC. Herbal soup consumption was associated with decreased NPC risk, with HRs of 0.31 (95% confidence interval (CI): 0.15–0.62) for the highest intake frequency and 0.29 (95% CI: 0.16–0.51) for a longer duration. However, herbal tea was not significantly associated. Moreover, we identified herbal soup was inversely associated with EBV seropositivity among all the participants at baseline, with the adjusted ORs being 0.78 (95% CI: 0.65–0.93) for immunoglobulin A antibodies against EBV capsid antigens (VCA-IgA) and 0.76 (95% CI: 0.64–0.91) for nuclear antigen 1 (EBNA1-IgA) in those with the highest frequency and 0.70 (95% CI: 0.59–0.84) for VCA-IgA and 0.64 (95% CI: 0.54–0.77) for EBNA1-IgA in those with the longer duration. Inverse associations were also observed in non-NPC individuals.ConclusionsWith inhibition of EBV reactivation by plants, herbal soup could significantly decrease the risk of NPC in endemic areas.

The Association of Single-Nucleotide Polymorphism rs13181 in ERCC2 with Risk and Prognosis of Nasopharyngeal Carcinoma in an Endemic Chinese Population.

ObjectiveWe examined whether the single-nucleotide polymorphism (SNP) rs13181 in the gene encoding excision repair cross complementation group 2 (ERCC2) is associated with the risk and prognosis of nasopharyngeal carcinoma (NPC).MethodsSNPs at rs13181 were genotyped in 439 NPC patients (NPC group) and 431 age- and gender-matched cancer-free controls (control group) from a region of China where NPC is endemic, and frequencies of GG, GT and TT genotypes were compared between the two groups in the case–control study. In a subset of 365 NPC cases, SNPs were examined for potential correlation with tumor-free survival time (TFS) and overall survival (OS).ResultsRelative to NPC risk with a TT genotype, NPC risk was similar with GT + GG genotypes (OR 1.052, 95% CI 0.656–1.688), after adjusting for gender, age, smoking history, and immunoglobin A against Epstein-Barr virus capsid antigen (EBV-VCA-IgA) status. Univariate analysis showed that the GG or GT genotype was associated with significantly worse TFS (p<0.001) and OS (p=0.010) than the TT genotype. Prognosis was significantly worse for men than for women (TFS, p=0.045; OS, p=0.031), for T3–T4 classification than for T1–T2 (TFS, p=0.009; OS, p=0.007), for N3 than for N0+N1+N2 (TFS, p<0.001; OS, p<0.001). Based on multivariate analysis, independent risk factors for poor TFS were GG or GT genotype (HR 2.629, 95% CI 1.625–4.254, p<0.001), T3–T4 classification (HR 2.146, 95% CI 1.244–3.701, p=0.006) and N3 (HR 2.527, 95% CI 1.574–4.059, p<0.001). GG or GT genotype (HR 2.217, 95% CI 1.283–3.832, p=0.004), gender (HR 1.989, 95% CI 1.046–3.785, p=0.036), T3–T4 (HR 2.431, 95% CI 1.306–4.526, p=0.005) and N3 (HR 2.693, 95% CI 1.637–4.432, p<0.001) were independent risk factors for poor OS.ConclusionThe rs13181 SNP in ERCC2 does not appear to be associated with NPC risk, but it may serve as an independent prognostic factor for NPC recurrence and death.

Monozygous twins with Epstein-Barr infection in the nasopharynx

The article describes a clinical case of monozygotic twins with hypertrophy of the organs of the lymphopharyngeal ring. The examination revealed IgG to the Epstein-Barr virus capsid antigen in both patients. One twin had Epstein-Barr virus DNA in the sample from the oropharynx. It was this child who had complaints typical of patients with adenoid vegetation, and he required surgical treatment (endoscopic shaver adenotomy and radio wave bilateral tonsillotomy). At the time of the initial examination and in the absence of complaints the second twin revealed no DNA of Epstein-Barr virus in the oropharynx. This patient was diagnosed with adenoids of 1-2 degree. Due to the complaints of the second twin that appeared during the following year of observation, with previously negative results on active Epstein-Barr virus markers served as the reason for a repeated specific laboratory examination. Epstein-Barr virus DNA was found in a sample from this child’s oropharynx. This case using the twin control method by partner enabled us to demonstrate the significance of Epstein-Barr virus in the pathogenesis and clinic manifestations of chronic diseases of the lympho-pharyngeal ring organs in children.

A combination of two ELISA tests for nasopharyngeal carcinoma screening in endemic areas based on a case-control study.

For populations with a high risk of nasopharyngeal carcinoma (NPC) in Guangdong province in southern China, mass screening is the first choice to prevent death from NPC. To improve the performance of NPC screening, we used a combination based on the IgA antibody against the Epstein-Barr virus (EBV) capsid antigen (VCA-IgA) and the IgA antibody against Epstein-Barr virus nuclear antigen 1 (EBNA1-IgA) to NPC screening by enzyme-linked immunosorbent assay (ELISA). A multiplication model was applied to measure the level of the combination. We evaluated the NPC screening effect of the markers.A case-control study was performed to assess the NPC screening effect of the markers. A total of 10,894 serum specimens were collected, including 554 samples from NPC patients and 10,340 samples from healthy controls. In the training stage, 640 subjects were randomly selected, including 320 NPC cases and 320 healthy controls. In the verification stage, 10,254 subjects were used to verify the NPC screening effect of the combination. Receiver operating characteristic (ROC) analysis was performed. In the verification stage, the combination achieved an sensitivity of 91.45%, a specificity of 93.45%, and an area under the ROC curve (AUC) of 0.978 (95% CI [0.968–0.987]). Compared with VCA-IgA and EBNA1-IgA individually, the combination had an improved screening performance. A probability (PROB) calculated by logistic regression model based on VCA-IgA and EBNA1-IgA was applied to NPC screening by ELISA in China. The AUC of the combination was a little bit larger than the PROB. There was a slight increase (3.13%) in the sensitivity of the combination compared to the sensitivity of the PROB, while the specificity was lower for the combination (92.50%) than for the PROB (95.94%). We successfully applied a combination of two ELISA tests based on VCA-IgA and EBNA1-IgA for NPC screening by using a multiplication model. The results suggested that the combination was effective and can be an option for NPC screening.

A Randomized Controlled Trial on Evaluation of Plasma Epstein-Barr Virus Biomarker for Early Diagnosis in Patients With Nasopharyngeal Carcinoma.

Early diagnosis of nasopharyngeal carcinoma (NPC) remains a major problem in Southern China. Epstein-Barr virus (EBV) biomarkers have been widely used in NPC screening. This study aims to evaluate the early diagnostic performances of individual EBV biomarkers in NPC. The levels of EBV biomarkers-IgA antibodies against EBV nuclear antigen 1 (EBNA1-IgA), EBV capsid antigen (VCA-IgA), EBV early antigen (EA-IgA), EBV BZLF1 transcription activator protein (Zta-IgA) and IgG antibodies against EBV BRLF1 transcription activator protein (Rta-IgG)-from 106 NPC patients (stage I and II) and 150 normal subjects were measured. VCA-IgA and EA-IgA were detected by immunofluorescence assay (IFA), EBNA1-IgA, Rta-IgG and Zta-IgA were measure by enzyme-linked immunosorbent assay (ELISA), and EBV DNA was detected by qPCR. Statistical analyses of a single index were conducted to evaluate the significance of NPC early diagnosis and TNM classification. The level of EBNA1-IgA, EBV DNA, VCA-IgA, EA-IgA, Rta-IgG and Zta-IgA in early-stage NPC was significantly higher than in healthy controls (all P < 0.001). EBNA1-IgA yielded the biggest area under the curve (AUC) of 0.962 in distinguishing early-stage NPC patients from the normal subjects, with a sensitivity of 91.5% and a specificity of 98.7%. However, EBV biomarker levels were not associated with tumor size (all P > 0.050), whereas four biomarker levels (EBNA1-IgA, EBV DNA, VCA-IgA, EA-IgA) were related to lymph node metastasis (N0 and N1-2), among which EBNA1-IgA and EBV DNA showed good performance. Finally, high correlation was found between VCA-IgA and EA-IgA (r > 0.800). A single EBNA1-IgA exhibits excellent discrimination performance in early diagnosis of NPC and could become a promising marker for NPC screening.

Evaluation of Epstein-Barr Virus Indirect Immunofluorescence Assay Results

Aim: Epstein-Barr Virus (EBV) causes Infectious Mononucleosis (IM) and chronic active EBV infections, malignant diseases such as Burkitt’s lymphoma, nasopharyngeal carcinoma, and posttransplant lymphoproliferative disease. More than 90% of children are infected with EBV until the age of six. The aim of this study was to diagnose EBV infection using Indirect Immunofluorescence Assay (IIFA) method and to evaluate these patients clinically. Methods: The tests were studied by using EBV Indirect Immunofluorescence Assay (IIFA) method. A total of 247 patients, 186 adults, and 61 children, were included in the study. Results: Five (2.7%) of the adults were EBV-Capsid Antigen (EBV-CA) IgM positive, 175 (94.1%) were IgG positive and 6 (3.2%) were seronegative. 10 (16.4%) of the child patients were IgM positive, which is considered as an acute IM infection marker, whereas the child patients had a significantly higher IgM rate than adults (p &lt;0.001). 39 (63.9%) of the child patients were IgG positive and 12 (19.7%) of them were seronegative. The rate of IgG positivity in children was significantly lower than in adults (p &lt;0.001). There was no significant difference between the genders in terms of IgM and IgG positivity rates in both adults and children (p &gt;0.05 for each). Conclusion: These results suggest that the presence of an acute EBV infection should be considered when the patient has viral diseases with similar clinical picture especially for children.

Clinical Validity of Serum Antibodies to SARS-CoV-2 : A Case-Control Study.

The clinical utility of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies remains undefined. To determine the clinical validity and utility of SARS-CoV-2 antibodies. Case-control study. First month of testing for coronavirus disease 2019 (COVID-19) by using a nucleic acid amplification test (NAAT) on nasopharyngeal swabs at the Johns Hopkins Hospital, Baltimore, Maryland (11066 persons). Of the 11066 tested persons, 115 (1%) were hospitalized adults investigated for COVID-19. Clinical record review was performed to classify them into a COVID-19 case group (n= 60) or a non-COVID-19 control group (n= 55). The laboratory control groups comprised 513 persons not tested by NAAT: 160 healthy laboratory employees, 101 persons positive for IgG antibodies against Epstein-Barr virus capsid antigen, 215 positive for thyroperoxidase antibody, and 37 positive for rheumatoid factor. Serum IgG and IgA antibodies against SARS-CoV-2 spike protein were detected by using enzyme-linked immunosorbent assay. Sensitivity and specificity of the SARS-CoV-2 IgG assay were 0.976 (95% CI, 0.928 to 0.995) and 0.988 (CI, 0.974 to 0.995), respectively, when performed 14 days or later after symptom onset, but sensitivity decreased at earlier time points. Immunoglobulin G developed rapidly and was sustained at high levels throughout follow-up (up to 58 days). Antibodies to SARS-CoV-2 predicted the odds of developing acute respiratory distress syndrome, which increased by 62% (CI, 48% to 81%; P< 0.001) for every 2-fold increase in IgG. Of 11066 NAAT-tested patients, 457 were repeatedly NAAT-negative, and serum samples were obtained for 18 such patients (6 COVID-19 case patients and 12 non-COVID-19 control patients). Antibodies were present in 5 of 6 case patients and none of the 12 control patients (P= 0.001). The study was retrospective and performed at a single center; the sample was small; follow-up was limited; and selection bias may have occurred. Antibodies to SARS-CoV-2 demonstrate infection when measured at least 14 days after symptom onset, are associated with clinical severity, and provide valuable diagnostic support in patients who test negative by NAAT but remain clinically suspicious for COVID-19. Clinical Immunology Laboratory, Department of Pathology, Johns Hopkins Hospital.