Genotypes of Epstein-Barr virus (EBV1/EBV2) in individuals with infectious mononucleosis in the metropolitan area of Belém, Brazil, between 2005 and 2016

Abstract Two types of Epstein Barr virus (EBV1/EBV2) have been shown to infect humans. Although their genomes are similar, the regions containing the EBNA genes differ. This study aimed to characterize the EBV genotypes of infectious mononucleosis (IM) cases in the metropolitan region of Belém, Brazil, from 2005 to 2016. A total of 8295 suspected cases with symptoms/signs of IM were investigated by infectious disease physicians at Evandro Chagas Institute, Health Care Service, from January 2005 to December 2016. Out of the total, 1645 (19.8%) samples had positive results for EBV by enzyme immunoassay and 251 (15.3%) were submitted to polymerase chain reaction (PCR) technique, using the EBNA3C region, in order to determine the type of EBV. Biochemical testing involving aspartate aminotransferase, alanine aminotransferase and gamma-glutamyl transferase were also performed. EBV type was identified by PCR in 30.3% (76/251) of individuals; of those, 71.1% (54/76) were classified as EBV1, 17.1% (13/76) as EBV2, and 11.8% (9/76) as EBV1+EBV2. The main symptoms/signs observed with EBV1 infection were cervical lymphadenopathy (64.8%, 35/54), fever (63%, 34/54), headache (20.4%, 11/54), arthralgia (20.4%, 11/54), and exanthema (18.5%, 10/54). EBV2 infection was detected in all but two age groups, with an average age of 24 years. The most common signs/symptoms of EBV2 were fever (76.9%, 10/13), average duration of 18 days, and lymphadenopathy (69.2%, 9/13). In contrast, EBV1+EBV2 coinfections were more frequent in those aged five years or less (20.0%, 2/10). The symptoms of EBV1+EBV2 coinfection included fever (66.7%, 6/9), and cervical lymphadenopathy and headache (33.3%, 3/9) each. The mean values of hepatic enzymes according to type of EBV was significantly different (p<0.05) in those EBV1 infected over 14 years of age. Thus, this pioneering study, using molecular methods, identified the EBV genotypes in 30.3% of the samples, with circulation of EBV1, EBV2, and EBV1+EBV2 co-infection in cases of infectious mononucleosis in the northern region of Brazil.

Role of combining EBNA assay and Bamh1-W assay in detection of EBV DNA loads in NPC / 实用医学杂志

Objective To investigate the role of real-time PCR(qPCR) assay for EBNA fragments in quantitative detection of Epstein-Barr virus(EBV)DNA loads and the diagnose value of combining EBNA assay with common qPCR assay (designed on Bamh1-W fragments) in Nasopharyngeal Carcinoma (NPC) patients Methods EBV DNA loads of 234 blood samples(66 NPC samples included)were detected using two methods and DNA loads inside and outside cells were detected respectively. Positive rate obtained through different methods was compared. Regression analysis and t test were used to validate the methodology. Results Positive rate of EB-NA assay(53.42% in all samples and 51.52% in NPC samples)was lower than that of Bamh1-W assay(69.23%in all samples ,71.21% in NPC samples),however the combination of two methods could enhance the positive rate(70.94% in all samples,72.73% in NPC samples),especially in NPC samples. The correlation R2 of EBNA assay and Bamh1-W assay was 0.577(P < 0.05)and the difference was statistically significant. In NPC samples , R2 was 0.828 (P > 0.05) and it showed good correlation but the difference was not statistically significant. Conclusions The combination of EBNA assay and Bamh1-W assay can improve the positive rate in EBV DNA loads detection and its efficiency is more significant in NPC patients ,which shows significance in EBV DNA loads quantification and in the auxiliary diagnosis of NPC.

Role of combining EBNA assay and Bamh1-W assay in detection of EBV DNA loads in NPC / 实用医学杂志

Objective To investigate the role of real-time PCR(qPCR) assay for EBNA fragments in quantitative detection of Epstein-Barr virus(EBV)DNA loads and the diagnose value of combining EBNA assay with common qPCR assay (designed on Bamh1-W fragments) in Nasopharyngeal Carcinoma (NPC) patients Methods EBV DNA loads of 234 blood samples(66 NPC samples included)were detected using two methods and DNA loads inside and outside cells were detected respectively. Positive rate obtained through different methods was compared. Regression analysis and t test were used to validate the methodology. Results Positive rate of EB-NA assay(53.42% in all samples and 51.52% in NPC samples)was lower than that of Bamh1-W assay(69.23%in all samples ,71.21% in NPC samples),however the combination of two methods could enhance the positive rate(70.94% in all samples,72.73% in NPC samples),especially in NPC samples. The correlation R2 of EBNA assay and Bamh1-W assay was 0.577(P < 0.05)and the difference was statistically significant. In NPC samples , R2 was 0.828 (P > 0.05) and it showed good correlation but the difference was not statistically significant. Conclusions The combination of EBNA assay and Bamh1-W assay can improve the positive rate in EBV DNA loads detection and its efficiency is more significant in NPC patients ,which shows significance in EBV DNA loads quantification and in the auxiliary diagnosis of NPC.

Expression of EBNA-1 in EB virus-induced lymphomas / 实用医学杂志

Objective To compare the different expression of EBNA-1 in the normal human lymphocytes and in the EBV-induced lymphomas so as to provide the basis for studying the mechanism of the EB virus-induced lymphoma. Methods Animal model of EBV-induced lymphoma was constructed in SCID mice. EBNA-1 gene expression level was detected by real-time PCR and Western blot. Results EBV-induced lymphomas in animal model were replicated. EBNA-1 gene was expressed both in normal human lymphoma cells and 3 EBV-induced lymphoma cells. In EBV-induced lymphoma cells , the expression was up-regulated 11 683 times than in normal lymphocytes. Western blot results showed that the expression of EBNA-1 protein in induced lymphoma cells increased significantly compared with that in normal lymphocytes. Conclusions EBNA-1 expression in EBV-induced lymphoma cells is up-regulated than in normal lymphocytes. EBNA-1 may play an important role in the development of EBV-induced lymphoma.

Expression of Epstein-Barr Virus Gene and Clonality of Infiltrated T Lymphocytes in Epstein-Barr Virus-associated Gastric Carcinoma

BACKGROUND: Epstein-Barr virus associated gastric lymphoepithelioma-like carcinoma (LELC) is characterized by the intensive infiltration of lymphoid cells, the presence of EBV, and the better prognosis over typical adenocarcinoma. Thus, it was assumable that viral latent proteins may be responsible for the recruitment of a certain T cell repertoire to EBV-associated gastric carcinoma. METHODS: To examine above possibility, EBV gene expression in gastric carcinoma tissues and usage of TCR among the tumor infiltrating lymphocytes were analyzed. RESULTS: EBV specific DNA and EBERs RNA were detected in 4 out of 30 patients. RT-PCR analysis revealed that all 4 of EBV-positive tumor tissues expressed EBNA1 mRNA and BARTs and LMP2a was detected only one sample out of 4. However, the EBNA2 and LMP-1 transcripts were not detected in these tissues. CD8+ T cells were the predominant population of infiltrating lymphocytes in the EBV-positive gastric carcinoma. According to spectra type analysis of infiltrating T cells, 10 predominant bands were detected by TCR Vbeta CDR3 specific RT-PCR from 4 EBV-positive tumor tissues. Sequence analysis of these bands revealed oligoclonal expansion of T cells. CONCLUSION: These findings suggest that clonally expanded T cells in vivo might be a population of cytotoxic T cells reactive to EBV-associated gastric carcinoma.

Advance on subtypes/variant of Epstein-Barr virus and relationship between subtypes/variants andEpstein-Barr virus related diseases / 国际儿科学杂志

Epstein-Barr virus( EBV )is a ubiquitous human gamma-1 herpesvirus,which is associated with human malignancies,such as nasopharyngeal carcinoma,Burkitt's lymphoma and Hodgkin lymphoma.Based on the polymorphism of amino acids in the polymorphic region of EBV genome,it can be classified into different subtypes/variants.By now,whether the subtypes/variants of EBV preferentially is associated with particular malignancies or represent geographical polymorphism remains controversial.This review summarized the literature on sequence variation in EBV genes,focusing on LMP-1,EBNA-1,and BZLF-1 and their distribution by geography and disease.

Polymerase chain reaction genotyping of Epstein-Barr virus in scraping samples of the tongue lateral border in HIV-1 seropositive patients

The Epstein-Barr virus (EBV) is the etiological agent of oral hairy leukoplakia (OHL), an oral lesion with important diagnostic and prognostic value in acquired immunodeficiency disease syndrome. The two EBV genotypes, EBV-1 and EBV-2, can be distinguished by divergent gene sequences encoding the EBNA-2, 3A, 3B, and 3C proteins. The purpose of this study was to identify the EBV genotype prevalent in 53 samples of scrapings from the lateral border of the tongue of HIV-1 seropositive patients, with and without OHL, and to correlate the genotypes with presence of clinical or subclinical OHL with the clinic data collected. EBV-1 and EBV-2 were identified through PCR and Nested-PCR based on sequence differences of the EBNA-2 gene. EBV-1 was identified in the 31 samples (15 without OHL, 7 with clinical OHL and 9 with subclinical OHL), EBV-2 in 12 samples (10 without OHL, 1 with clinical and 1 subclinical OHL), and a mixed infection in 10 samples (2 without OHL, 3 with clinical and 5 with subclinical OHL). The presence of EBV-1 was higher in women, but a significant statistical result relating one the EBV genotypes to the development of OHL was not found. We conclude that the oral epithelium in HIV-1 seropositive patients can be infected by EBV-1, EBV-2 or by a mixed viral population.

Epstein-Barr virus nuclear antigen-2 detection and typing in immunocompromised children correlated with lymphoproliferative disorder biopsy findings

Epstein-Barr virus (EBV), the causative agent of infectious mononucleosis, plays a significant role as a cofactor in the process of tumorigenesis, and has consistently been associated with a variety of malignancies especially in immunocompromised patients. Forty-four children and adolescents (21 liver transplant patients, 7 heart transplant, 5 AIDS, 3 autoimmune hepatitis, 2 nephritic syndromes, 2 medullar aplasia, 2 primary immunodeficiency disorder patients, 1 thrombocytopenic purpura and 1 systemic lupus erythematosus) presenting with chronic active EBV infection (VCA-IgM persistently positive; VCA-IgG > 20 AU/mL and positive IgG _ EBNA) had peripheral blood samples obtained during clinically characterized EBV reactivation episodes. DNA samples were amplified in order to detect and type EBV on the basis of the EBNA-2 sequence (EBNA2 protein is essential for EBV-driven immortalization of B lymphocytes). Although we have found a predominance of type 1 EBNA-2 virus (33/44; 75 percent), 10 patients (22.73 percent) carried type 2 EBNA-2, and one liver transplant patient (2.27 percent) a mixture of the two types, the higher proportion of type 2 EBV, as well as the finding of one patient bearing the two types is in agreement with other reports held on lymphoproliferative disorder (LPD) patients, which analyzed tumor biopsies. We conclude that EBNA-2 detection and typing can be performed in peripheral blood samples, and the high prevalence of type 2 in our casuistic indicates that this population is actually at risk of developing LPD, and should be monitored.

Expression of LMP-1、EBNA-2 in the Follicular Epithelium Neoplasm of the Thyroid / 医学研究杂志

0.05);LMP-1 and EBNA-2 in papillary thyroid carcinoma had no correlation.Conclusion Some thyroid papillary carcinomas may be related to EB virus infection,the relationship has yet to be further studied.

Clinical Significance of Epstein-Barr Virus Expression in Breast Cancer / 한국유방암학회지

PURPOSE: Epstein-Barr Virus (EBV) is well understood as an oncogenic virus in human tumors. Its association with breast cancers has been reported but is still in controversy. So we have examined the expression of EBV in breast cancers and evaluated the relationship between the well-known prognositc factors of breast cancer and EBV expression. METHODS: A retrospective study was conducted with patients who had been re-evaluated to confirm the diagnosis based on immunohistochemical analysis with EBNA-2 expression, between January 1991 and December 2002. The cases were assigned to the positive lesion that displayed 10% or more of immunoreactive cells. RESULTS: The expressions of EBNA-2(Ebstein Barr virus nuclear antigen - 2) were noted in 26 (21.1%) out of 123 cases of breast cancer patients and 4 (20%) out of 20 cases in a control group of benign tumors. The expression of EBV in breast cancers and that of a control group were not different significantly. But, the correlation between the expression of EBNA-2 and ER status was noted statistically significant (P=0.040). CONCLUSION: Judging from the results of our study, EBV infection detected in breast cancer seems to be latent and the association of EBV to breast cancer is less likely related.

Detection and Subtyping of Epstein-Barr Virus in Gastrointestinal Adenocarcinomas and Malignant Lymphomas

Epstein-Barr virus (EBV) has been linked to a spectrum of neoplastic conditions, including Burkitt's lymphoma, nasopharyngeal carcinoma, Hodgkin's disease, lymphoepithelioma-like carcinomas and malignant lymphomas in immunocompromised state. To determine the prevalence and the subtype of EBV in gatrointestinal malignancies, fifty cases of adenocarcinomas and seventeen cases of malignant lymphomas were analyzed by EBERs in situ hybridization and polymerase chain reaction using primers for EBNA-1, EBNA-2A and EBNA-2B, on the paraffin sections. In addition, immunohistochemical stain for p53 protein was performed to investigate the potential role of EBV infection on tumor suppressor gene, p53, during tumorigenesis. EBER was detected in 6 of 26 gastric adenocarcinomas, 2 of 24 colon adenocarcinomas, and 8 of 17 malignant lymphomas. EBER was more prevalent in malignant lymphoma arising in the intestine (6/6) than in the stomach (2/11), and was detected in both B and T cell phenotypes. EBNA-1 was positive in 11 of 16 EBER positive cases and the subtyping was possible in 8; both type 1 and 2 were detected in gastric cancers, whereas only type 2 was found in intestinal neoplasms. In adenocarcinomas the high rate of p53 protein overexpression was found in both EBER positive (8/8) and negative cases (32/42), whereas the positive rate was higher in EBER positive cases (7/8) than in EBER negative cases (4/9) of malignant lymphomas. From the results, it can be concluded that EBV infection and the p53 tumor suppressor gene are independently associated in a significant portion of the gastrointestinal malignancies, but the mechanism of action remains to be elucidated.