Epstein-Barr Virus Type Research Articles

The role of herpesviruses in endometrial pathology: frequency of herpesvirus DNA detection in the endometrium, cervix and vagina in patients with chronic endometritis and endometrial hyperplasia

Introduction. Chronic endometritis (CE) is reported in 12% of patients with primary infertility. Herpesviruses are considered as one of the causes of CE.The aim of the study. To evaluate the frequency of herpesviruses DNA detection in endometrium, cervix and vagina in patients with chronic endometritis and endometrial hyperplasia.Materials and methods. 167 women of reproductive age were included in the study. Samples from the posterolateral wall of the vagina, cervix and endometrium were taken from each patient. The obtained samples were analyzed by real-time PCR for the presence of Herpes simplex viruses type 1 and 2 (HPV 1,2), Epstein-Barr virus (EBV), Cytomegalovirus (CMV) and Human herpesvirus type 6 (HHV6). Endometrial tissue was subjected to morphologic examination.Results. Depending on the morphology of endometrial biopsy, all patients were divided into three groups: with CE (N=53), with endometrial hyperplasia (EH, N=63), with normal endometrium (N=51). There were no statistically significant differences in the frequency of herpesvirus detection in endometrial, cervical, and vaginal samples. In endometrial samples, EBV was detected in 1 (1.9%) patient with CE (1.9%) and 1 (1.6%) patient with EH, HPV1,2 — in 1 (2%) woman with normal endometrial histology; DNA of other herpesviruses was not detected.Conclusions. The relationship between herpesvirus type, positive locus and the presence of endometrial pathology has not been established. The frequency of herpesvirus DNA detection in endometrial, cervical, and vaginal samples was not significantly different between the groups of women with CE, EH, and normal endometrial histology. Only 2 (1.7%) of endometrial samples with pathologic morphology were positive for DNA of herpesviruses, in both cases it was EBV.

Hitting Epstein Barr virus where it hurts: computational methods exploration for siRNA therapy in alleviating Epstein Barr virus-induced multiple sclerosis.

Multiple sclerosis (MS), an intricate neurological disorder, continues to challenge our understanding of the pivotal interplay between the immune system and the central nervous system (CNS). This condition arises from the immune system's misdirected attack on nerve fiber protection, known as myelin sheath, alongside nerve fibers themselves. This enigmatic condition, characterized by demyelination and varied clinical manifestations, prompts exploration into its multifaceted etiology and potential therapeutic avenues. Research has revealed a potential connection between Epstein Barr virus (EBV), specifically Epstein Barr Nuclear Antigen 1 (EBNA-1), and MS. The immune response to EBNA-1 antigen triggers the production of anti-EBNA-1 molecules, including IgG that identify a similar amino acid sequence to EBNA-1 in myelin, inadvertently targeting myelin sheath and contributing to MS progression. Currently, no treatment exists for EBNA-1-induced MS apart from symptom management. Addressing this, a novel potential therapeutic avenue utilizing small interference RNAs (siRNA) has been designed. By targeting the conserved EBNA-1 gene sequences in EBV types 1 and 2, five potential siRNAs were identified in our analysis. Thorough evaluations encompassing off-target binding, thermodynamics and secondary structure elucidation, efficacy prediction, siRNA-mRNA sequence binding affinity exploration, melting temperature, and docking of siRNAs with human argonaute protein 2 (AGO2) were conducted to elucidate the siRNAs efficiency. These designed siRNA molecules harnessed promising silencing activity in the EBNA-1 gene encoding the EBNA-1 antigen protein and thus have the potential to mitigate the severity of this dangerous virus.

Investigating the role of Epstein-Barr virus and human papillomavirus types 16 and 18 co-infections in cervical disease of Iranian women.

High-risk human papillomaviruses (HR-HPVs) are known to contribute to cervical cancer (CC), but the role of Epstein-Barr virus (EBV) in this process remains unclear, despite EBV's widespread detection in premalignant and malignant cervical tissues. In this cross-sectional study of 258 cervical samples, including both formalin-fixed paraffin-embedded (FFPE) and fresh cervical tissues, the presence and viral load of HR-HPVs (HPV-16 and HPV-18) and EBV were evaluated in Iranian women with cervical intraepithelial neoplasia (CIN), squamous cell carcinoma (SCC), and a cervicitis control group using real-time PCR. The study revealed a significant correlation between disease severity and both increased HPV-16 positivity and HPV-16 and HPV-18 co-infection (p<0.001). Interestingly, the control group had a higher frequency of EBV-positive cases than SCC/CIN groups (p<0.001). HPV-16 DNA load increased with disease severity (P<0.001), while HPV-18 showed no significant difference (P=0.058). The control group had a higher EBV DNA load compared to SCC/CIN groups (P=0.033). HPV-16 increased the risk of CIN II, CIN III, and SCC, while HPV-18 increased the risk of CIN II and CIN III. Notably, EBV was associated with a lower risk of CIN groups and SCC. No significant difference in EBV co-infection with HPV-16/18 was found, failing to support the hypothesis that EBV is a cofactor in CC. However, high EBV viral load in the control group suggests a potential "hit and run hypothesis" role in CC progression. This hypothesis suggests that EBV may contribute briefly to the initiation of CC with an initial impact but then becomes less actively involved in its ongoing progression.

Identification of Epstein-Barr virus after topical treatment for oral hairy leukoplakia: A preliminary study.

This study evaluated the presence of Epstein-Barr virus type 1 (EBV-1) DNA in patients living with HIV, before and after three different topical therapy protocols for oral hairy leukoplakia (OHL). The sample consisted of five patients treated with topical solution of 25% podophyllin resin; six with 25% podophyllin resin plus 5% acyclovir cream; and four with 25% podophyllin resin plus 1% penciclovir cream. DNA was extracted from OHL scrapings and amplified by the PCR using specific primers for EBV-1 (EBNA-1). Clinical healing of OHL lesions was observed across all treatment groups over time. At baseline, EBNA-1 was detected in all OHL lesions. After treatment, OHL samples from three patients treated with 25% podophyllin resin plus 5% acyclovir cream and from one patient treated with 25% podophyllin resin plus 1% penciclovir cream exhibited negative EBNA-1 viral gene encoding. Despite the clinical resolution of OHL, 11 patients (73.3%) showed EBNA-1 positivity immediately after the lesion disappeared. Three patients (20%) treated with podophyllin resin displayed both EBNA-1 positivity and a recurrence of OHL, in contrast to no recurrence in the other two groups. These findings suggest potential associations between treatment formulations, EBNA-1 persistence, and the recurrence of OHL lesions.

Abstract LB412: BRAFp.G469E hotspot mutation promotes nasopharyngeal carcinoma tumorigenesis with potential druggability

Abstract Nasopharyngeal Carcinoma (NPC) is a distinct type of Epstein-Barr Virus (EBV)-associated head &amp; neck cancer. It is most prevalent in Southeast Asia with a high incidence rate of ~15/100,000. While mutations of the MAPK pathways are found in nearly one-fifth of head &amp; neck squamous cell carcinoma (HNSCC) in Western countries (TCGA-HNSCC Firehose cohort, N=512), and some of which are druggable in HNSCC setting (e.g. MAPK1 and HRAS mutations), we sought to determine the cumulative mutational rate of this potentially druggable pathway in 164 micro-dissected NPC cases using recently published next-generation sequencing data (98 whole-exome and 63 whole-genome-sequenced cases). An overall rate of 9.32% cases of NPC harbored MAPK pathway mutations (16 mutations in 15 tumors). Four genes are recurrently mutated: 4 NRAS (all Q61R/L hotspot mutations), 3 HRAS (G12S, Q61R, S106L), 2 BRAF (R726H, Y647*), and 2 SHC4 (Q519R, A552T) mutations. Among which, both patients bearing BRAF mutations died of NPC, suggestive of the biological importance of BRAF mutations in NPC progression. Prompted by an independent clinical report on an advanced NPC patient with somatic co-mutations of BRAFp.G469E and KRASp.G12D being sensitive to the B-raf/C-raf inhibitor sorafenib, we first examined the biological functions and potential druggability of this particular BRAFp.G469E mutation in NPC, as it represents a hotspot mutation site across 20 cancer types (BRAFp.G469E/A/R/V/* found in 137/70655 pan-cancer samples, cBioPortal). Based on the NPC clinical report, we hypothesized that this BRAFp.G469E hotspot mutation could drive NPC tumorigenesis and might be potentially sensitive to the B-raf inhibitor sorafenib. By ectopic expression of BRAFp.G469E mutation, BRAF-wildtype (wt) and EGFP-control gene into a recently developed Epstein-Barr virus-positive nasopharyngeal carcinoma [EBV(+)NPC] cell line, NPC43, we determined the biological functions of this mutation in NPC. We showed that NPC43-BRAFp.G469E-expressing cells significantly drove a 4-fold increase in 3D tumoroid growth vs. BRAF-wt and EGFP-control cells on day 7 (flat-bottom low attachment plates). Further, wound healing assay demonstrated a significantly higher migration rate of NPC43-BRAFp.G469E cells achieving ~80% wound healing (at 96h) vs. 30% and 10% healing in BRAF-wt and EGFP-control cells. Lastly, our preliminary drug study showed that NPC43-BRAFp.G469E cells displayed higher sensitivity to low dose sorafenib (33.3nM and 100nM) than BRAF-wt and EGFP-control cells, achieving ~20% and 30% growth inhibition at 72h, vs. no growth inhibition in control cells. Thus, BRAFp.G469E hotspot mutation can drive NPC tumorigenesis, and may confer sensitivity to raf inhibitors such as sorafenib in NPC. Potential druggability of other BRAF mutations in NPC should be investigated. Acknowledgements: The study was supported by Research Impact Fund (R4015-19F), Research Grants Council, University Grants Committee, Hong Kong SAR during 6/29/20-9/7/21. Citation Format: Chun Ho Law, Zoey Wing Yee Liu, Franky Leung Chan, Tin Yu Samuel Law, Vivian W.Y. Lui. BRAFp.G469E hotspot mutation promotes nasopharyngeal carcinoma tumorigenesis with potential druggability [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB412.

Epstein-Barr Virus DNA Is Associated With Conjunctival Squamous Cell Carcinomas: A Case-Control Study From Zimbabwe.

Incidence of conjunctival squamous cell carcinoma (cSCC) in Zimbabwe is >30-fold higher than the global average. cSCC risk is notably higher among people with human immunodeficiency virus, implicating impaired immune response and a yet unknown infectious etiology. Formalin-fixed, paraffin-embedded blocks from Zimbabwe, comprising conjunctival precancer (n = 78), invasive cSCC cases (n = 148) and nonmalignant eye lesions (n = 119), were tested for multiple DNA viruses using Luminex bead-based technology. Epstein-Barr virus (EBV) type 1 positivity was strongly associated with cSCC diagnosis (adjusted odds ratio [aOR], 5.6 [95% confidence interval {CI}, 3.0-10.4) and marginally associated with precancer (aOR, 2.1 [95% CI, 1.0-4.5]). On analyzing EBV transcriptional activity with any of LMP1, EBNA1, and BZLF1, RNA transcripts were detected in 5 of 112 controls, 3 of 67 precancers, and 10 of 139 cases and none were associated with conjunctival case status. Our EBV DNA data suggest that EBV may play a role in cSCC. However, the low detection rate of EBV RNA supports further investigation to infer causality.

Shared and distinct interactions of type 1 and type 2 Epstein-Barr Nuclear Antigen 2 with the human genome

BackgroundThere are two major genetic types of Epstein-Barr Virus (EBV): type 1 (EBV-1) and type 2 (EBV-2). EBV functions by manipulating gene expression in host B cells, using virus-encoded gene regulatory proteins including Epstein-Barr Nuclear Antigen 2 (EBNA2). While type 1 EBNA2 is known to interact with human transcription factors (hTFs) such as RBPJ, EBF1, and SPI1 (PU.1), type 2 EBNA2 shares only ~ 50% amino acid identity with type 1 and thus may have distinct binding partners, human genome binding locations, and functions.ResultsIn this study, we examined genome-wide EBNA2 binding in EBV-1 and EBV-2 transformed human B cells to identify shared and unique EBNA2 interactions with the human genome, revealing thousands of type-specific EBNA2 ChIP-seq peaks. Computational predictions based on hTF motifs and subsequent ChIP-seq experiments revealed that both type 1 and 2 EBNA2 co-occupy the genome with SPI1 and AP-1 (BATF and JUNB) hTFs. However, type 1 EBNA2 showed preferential co-occupancy with EBF1, and type 2 EBNA2 preferred RBPJ. These differences in hTF co-occupancy revealed possible mechanisms underlying type-specific gene expression of known EBNA2 human target genes: MYC (shared), CXCR7 (type 1 specific), and CD21 (type 2 specific). Both type 1 and 2 EBNA2 binding events were enriched at systemic lupus erythematosus (SLE) and multiple sclerosis (MS) risk loci, while primary biliary cholangitis (PBC) risk loci were specifically enriched for type 2 peaks.ConclusionsThis study reveals extensive type-specific EBNA2 interactions with the human genome, possible differences in EBNA2 interaction partners, and a possible new role for type 2 EBNA2 in autoimmune disorders. Our results highlight the importance of considering EBV type in the control of human gene expression and disease-related investigations.

Frequency and association of Epstein-Barr Virus genotype in rheumatoid arthritis patients of Khyber Pakhtunkhwa, Pakistan.

Rheumatoid arthritis (RA) is an immune-mediated, polyarthritis linked with various genetic and environmental causative agents. Among environmental triggers, Epstein-Barr Virus (EBV) is considered the most potent etiological agent. This study aimed to investigate the prevalence of EBV and its genotypes in RA patients and to investigate their association with clinical and laboratory parameters of RA. This study included blood samples of RA and control healthy individuals (100 each). Blood samples along with clinical and laboratory parameters were collected from patients after consent in the Department of Rheumatology, at Lady Reading Hospital, in Peshawar Pakistan. Blood samples were processed for DNA extraction followed by PCR amplification for EBV detection and genotype discrimination. RA patients were 85 females and 15 males with a mean age of 40.13±14.05 years. EBV Type-1 was detected in 45% of RA and 9% of control cases. The mean disease duration of RA patients was 6.61±6.23 years. Out of 100 diseased patients, 43% were seropositive rheumatoid arthritis (SPRA) and showed a significant correlation with a family history of RA in EBV-positive individuals (P = 0.017). The demographic, clinical, and laboratory parameters of RA patients showed a non-significant association with EBV. Moreover, only a family history and Serum creatinine of RA patients showed a significant association with EBV (P = 0.0001 and P = 0.022 respectively). It is concluded that EBV-1 is prevalent and associated with RA. Further investigation is required for detailed genetic analysis of EBV to determine its possible role in modulating the immune system in RA.

Molecular profile of Epstein–Barr virus in representatives of four ethnic groups of Russia: virus types, &lt;i&gt;LMP1&lt;/i&gt; gene variability and malignancies

Rationale. The discovery of two Epstein–Barr virus (EBV) types, EBV-1 and EBV-2, which differ in their transforming abilities stimulated their study in various populations in order to elucidate the relationship with malignant neoplasms.Aim. To study EBV-1 and EBV-2 in representatives of four ethnic groups of Russia, sequence analysis of the LMP1 oncogene in virus isolates, and search for a correlation between the types of EBV and the incidence of certain types of malignant tumors in four ethnic groups.Materials and methods. From total DNA obtained from oral washings of Adygeans, Kalmyks, Tatars and Slavs, representatives of 3 Russian republics and the Moscow region, EBV DNA was amplified; its presence and concentration were analyzed by nested PCR, and each product was examined for the virus type and LMP1 variant. Types of EBV found in representatives of In four ethnic groups were compared with the incidence rates of malignant tumors, among which EBV-associated cases may occur, in population of three republics and the Moscow region.Results. It was observed that higher gastric cancer and lymphomas morbidity among Tatarstan and Moscow region populations (compared to populations of Adygea and Kalmykia) correlated with predominance in these ethnic groups – Tatars and Slavs – EBV-1 type having transforming potential in vitro. In Adygeis, non-transforming EBV-2 type was prevalent, and Kalmyks were infected with both types in approximately equal proportion. however, differences between tumor morbidities between the compared groups were statistically insignificant (p &gt;0.05). The identified spectrum of LMP1 variants, as well as the results of their sequencing, show genetic relations between EBV strains circulating in the studied ethnic groups.Conclusion. Study of EBV isolates for the first time performed in 4 ethnic groups of this country showed differences in distribution of the types of this virus but there was no correlation with tumor morbidity.

Aggressive periodontitis in southwestern American Indian adolescents.

This study determined the prevalence of aggressive (molar-incisor pattern) (Ag/MI) periodontitis and assessed the associated subgingival bacterial-herpesvirus microbiota in Pueblo Indian adolescents in the southwestern United States. The study included 240 Pueblo Indian adolescents, aged 13-20years old, residing in three Rio Grande River villages in New Mexico and the Hopi Pueblo reservation in Arizona. Adolescents with Ag/MI periodontitis or periodontal health provided subgingival samples for culture of bacterial pathogens and for polymerase chain reaction detection of periodontal herpesviruses. Ag/MI periodontitis was detected in 22 (9.2%) Pueblo Indian adolescents, with 21 exhibiting a localized molar-incisor breakdown pattern. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and other red/orange complex bacterial pathogens predominated in Ag/MI periodontitis, whereas periodontal health yielded mainly viridans streptococci and Actinomyces species. Periodontal herpesviruses demonstrated a 3.5 odds ratio relationship with Ag/MI periodontitis. The only adolescent with generalized Ag/MI periodontitis harbored viral co-infection by cytomegalovirus plus Epstein-Barr virus Type 1, in addition to A. actinomycetemcomitans, P. gingivalis, and several other periodontopathic bacteria. Pueblo Indian adolescents showed an unusually high prevalence of early-age Ag/MI periodontitis predominated by periodontopathic bacteria and herpesviruses suspected to be major etiologic agents of the disease.

The infectious status of patients with optic neuritis of inflammatory and demyelinating etiologies

Purpose: to study the prevalence of various chronic infections, the frequency of their reactivation and characteristic associations of microorganisms in patients with optic neuritis of various etiologies.Material and methods. 13 patients with optic neuritis (ON) and 12 patients with ON and multiple sclerosis were tested for a broad range of infectious agents in their blood serum.Results. All patients were shown to have mixed infections; 64 % of them had an association of 4 or more infectious agents. The patients of both groups were infected with Epstein-Barr virus (100 %), Cytomegalovirus (92 %) and Herpex simplex viruses (92 %) types 1 and/or 2 with serological signs of their reactivation. The combination of Herpes virus reactivation, Toxoplasma and urogenital infection was more often detected in patients with demyelinating ON. In patients with multiple sclerosis, a mutual correlation was detected: the higher the degree of infection (combination of 5 or more pathogens), the lower the visual acuity at the onset of the disease (p &lt; 0.05) and the less favourable the vision prognosis.Conclusion. Possibly, mixed infection may play an important role in the pathogenesis of ON, including that of demyelinating etiology, as a trigger or an aggravating factor.

Prevalence of Epstein-barr Virus (EBV) among Patients with Oral Squamous Cell Carcinoma from Ahvaz, Iran: A Case-Control Study.

Epstein-Barr virus (EBV), one of the most significant causes of lymphoid and epithelial cancers, has been linked to oral carcinogenesis; however, this etiological association remains controversial. To investigate this association, the present study aimed to determine the prevalence of EBV in cancerous and non-cancerous oral tissues from Ahvaz, Iran. In total, 164 blocks of formalin-fixed paraffin-embedded tissues from oral squamous cell carcinoma (OSCC), including 76 tongue squamous cell carcinomas and 88 non-cancerous tongue tissues, were collected from Ahvaz Imam Khomeini Hospital, Ahvaz, Iran, from December 2014 to March 2019, for this case-control study. The tissues were cut into 15-μm-thick sections, and DNA was extracted using a solution of Phenol, Chloroform, and Isoamyl Alcohol. The EBV detection and typing were performed using nested polymerase chain reaction. The EBV was detected in 9 (5.48%) out of the 164 samples studied, including 4 (5.26%) of the 76 SCC cases and 5 (5.68%) of the 88 samples in the control group (P>0.05). The EBV was positive in 2.40% of the 83 male and 8.6% of the 81 female samples (P>0.05). In terms of the histological grades of the case group, 3 (3/57) and 1 (1/13) of the EBV-positive samples were well and moderately differentiated, respectively (P>0.05). For EBV typing, the 9 EBV-positive samples were tested, and it was found that 2 and 7 of the cases were EBV type I and II, respectively. Results of the current study demonstrated the low frequency of EBV in Iranian patients with OSCC, with EBV type II predominating. Further studies are required to clarify the association between EBV and OSCC.

Epstein-Barr virus (Herpesviridae: Lymphocryptovirus) types 1 and 2 and other viral markers in patients with nasopharyngeal carcinoma in two geographically and ethnically distinct regions of Russia

The discovery of two types of Epstein-Barr virus (EBV) (EBV-1 and EBV-2) that have different biological properties stimulated the search for neoplasms associated with each type of the virus. The aim of the work is to study the nature of the association of nasopharyngeal cancer (NPC) with EBV-1 and EBV-2, serological activity for each viral type and the concentration of EBV DNA in the blood plasma of two gender, age and ethnic groups of NPC patients that represent geographically and climatically different regions of Russia,. In the blood plasma of patients with NPC and other non- EBV associated tumors of oral cavity (OTOCEBV-) from the North Caucasian (NCFD) and Central (CFD) Federal Districts of Russia, the types of EBV and the concentration of viral DNA were determined using respectively «nested» and real time PCR; titers of IgG and IgA antibodies to viral capsid antigen (VCA) were measured in indirect immunofluorescence assay. The blood plasma samples testing showed that NPC and OTOCEBV- patients were infected with both types of EBV in approximately equal proportions. In two groups of NPC patients infected with one of the virus types only, EBV-1 or EBV-2, respectively, no statistically significant differences were found between the geometric mean values of IgG and IgA anti-EBV antibody titers and viral DNA concentrations in blood plasma. The distribution of virus types was not affected by either patient gender or ethnogeographic origin. The difference was found only between age groups: EBV-2 dominated in NPC patients up to 60 years, and EBV-1 was prevalent in patients over 60 years. The lack of the predominance of one of EBV types in NPC patients that are the representatives of different ethnic groups from geographically and climatically different regions, suggests that none of these factors play an important role in the NPC carcinogenesis. Evidently, each type of EBV, EBV-1 or EBV-2, if the necessary conditions arise, are able to exhibit its oncogenic potential to initiate tumor development.

Genotypes Distribution of Epstein-Barr Virus among Lymphoma Patients in Ethiopia.

Epstein-Barr virus (EBV) is an oncogenic herpes virus associated with several human malignancies. Two main EBV genotypes (type 1 and type 2) distinguished by the differences in EBV nuclear antigens are known. Geographic variability in these genetic differences has been observed in the incidence of some EBV-related tumors. Here, we investigated the genetic variation of EBV in lymphoma specimens collected in Ethiopia. A total of 207 DNA samples were used for EBV detection and typing, and EBNA1 and EBNA3C genes were used to detect and subtype the EBV genome, respectively. EBV genotype 1 was detected in 52.2% of lymphoma patients. EBV genotype 2 was detected in 38.2% of the lymphoma patients, and 9.7% were coinfected by both EBV genotypes. Overall, 52.8% of the Hodgkin's lymphoma (HL) patients and 51.8% of non-Hodgkin's lymphoma (NHL) patients showed the presence of genotype 1. Meanwhile, 42.8% and 2.3% of HL patients and 35.8% and 12.4% of NHL patients showed EBV genotype 2 and both genotypes, respectively. Significant associations between the age groups and EBV genotypes were observed (p = 0.027). However, no significant association was seen between EBV genotypes and other sociodemographic and clinical characteristics. This study showed that the distribution of EBV genotype 1 was higher in Ethiopian lymphoma patients.

Nasopharyngeal Carcinoma (NPC) in Russia and Epstein - Barr Virus Types: Analysis of the Influence of Gender, Ethnic and Age Factors on the Virus Types Spread in NPC Patients

Introduction: The discovery of two molecular types of Epstein-Barr virus, EBV-1 and EBV-2, with different biological properties, stimulated the search for new neoplasms associated with each type of the virus. Objective: The aim of this study was to analyze the prevalence of EBV-1 and EBV-2 in NPC patients from different climatic and geographical regions of Russia, representatives of different gender, ethnic and age groups of patients. Materials and Methods: Using nested PCR EBV types were determined in blood plasma of NPC and other non-EBV-associated tumors of the oral cavity (OTOC/EBV-) patients from the North Caucasus (NCFD) and the Central (CFD) Federal Districts of Russia. IgG and IgA antibody titers to viral capsid antigen (VCA) were determined by indirect immunofluorescence. Results: The studies carried out have shown that NPC and OTOC/EBV- patients were infected with both EBV types in roughly the same ratio. There were also no statistically significant differences in serological activities of NPC patients infected with EBV-1 or EBV-2. The prevalence of EBV-2 types 1 and 2 was not influenced by gender or ethno-geographic affiliation of NPC patients. The only difference was EBV-2 predominance in NPC patients under 60, and EBV-1 predominance in those over 60 years old. Conclusion: The absence of a predominance of one of the EBV types in NPC patients indicates that neither EBV-1 nor EBV-2 is strictly NPC-associated in Russia, and that different ethnic and climatic-geographical factors in the spread of virus types in NPC patients are not playing an essential role.

Distribution of Epstein-Barr Virus LMP1 Variants in Patients with Infectious Mononucleosis and Association with Selected Biochemical and Hematological Parameters.

The molecular diversity of Epstein-Barr virus (EBV) is exceptionally complex and based on the characterization of sequences coding for several viral genes. The aim of this study was to analyze the distribution of EBV types 1 and 2 and to characterize LMP1 variants in a cohort of 73 patients with infectious mononucleosis (IM), as well as to investigate a possible association between viral diversity and relevant clinical parameters. Population-based sequencing of EBNA-2 gene showed the presence of EBV type 1 in all IM patients. Analysis of LMP1 gene found a restricted repertoire of LMP1 variants with the predominance of wild-type B95-8, China1, Mediterranean and North Carolina variants with the presence of more than one LMP1 variant in 16.4% of patients. Co-infections with different LMP1 variants were associated with significantly higher levels of C-reactive protein and lower levels of maximal neutrophil counts and minimal platelet count. The results of this study have shown a narrow repertoire of LMP1 variants and an exclusive presence of EBV type 1 in a cohort of IM from Croatia, suggesting a characteristic local molecular pattern of this virus. The clinical importance of distinct immunobiological features of IM patients with LMP1 variant co-infections needs to be investigated further.

Epstein-Barr virus variation in people living with human immunodeficiency virus in southeastern China

BackgroundPatients infected with HIV are at high risk of developing Epstein-Barr Virus (EBV)-related diseases. The genotype and viral biological behavior of EBV infection in patients with human immunodeficiency virus-1 (HIV) in China remain unclear. This study analyzed the characteristics of EBV in patients infected with HIV in southeastern China.MethodsA total of 162 HIV-infected patients and 52 patients without HIV were enrolled in this study. EBV viral load in blood was determined by fluorescence quantitative PCR. EBV typing was performed using saliva according to polymorphisms in the EBNA3C region. EBV LMP-1 carboxy terminus (C-ter) was sequenced, and compared with the epidemic strains in the world.ResultsAmong HIV infected patients, the EBV strain variant was mainly EBV-1, while EBV-2 had a higher viral load than EBV-1 (P = 0.001) and EBV-1/2 (P = 0.002). HIV infected patients had higher active virus replication. The EBV LMP-1 variants were mainly the China1 variant. HIV-infected patients had different nucleic acid positions of 30-bp deletion (del30) and had a higher incidence of high 33-bp tandem repeats (rep33) copies than non-HIV-infected patients. There was a difference in the mutations of EBV LMP-1 C-ter del30 and ins15 between HIV infected patients and the control group (P < 0.001).ConclusionIn southeastern China, EBV in HIV-infected patients had higher active virus replication; EBV infection was mainly EBV-1, and EBV-2 infection has higher EBV virus load; hotspot mutations of LMP-1 C-ter were different between HIV-infected patients and non-HIV-infected patients.Trial registration: This study was approved by the ethics committee of the First Affiliated Hospital of Zhejiang University School of Medicine (Approval No. 2018764), and registered in Chinese Clinical Trial Registry on 3 June 2019 (ChiCTR, ChiCTR1900023600, http://www.chictr.org.cn/usercenter.aspx).

Prevalence of Epstein-barr Virus (EBV) among Patients with Oral Squamous Cell Carcinoma from Ahvaz, Iran: A Case-Control Study

Epstein-Barr virus (EBV), one of the most significant causes of lymphoid and epithelial cancers, has been linked to oral carcinogenesis; however, this etiological association remains controversial. To investigate this association, the present study aimed to determine the prevalence of EBV in cancerous and non-cancerous oral tissues from Ahvaz, Iran. In total, 164 blocks of formalin-fixed paraffin-embedded tissues from oral squamous cell carcinoma (OSCC), including 76 tongue squamous cell carcinomas and 88 non-cancerous tongue tissues, were collected from Ahvaz Imam Khomeini Hospital, Ahvaz, Iran, from December 2014 to March 2019, for this case-control study. The tissues were cut into 15-μm-thick sections, and DNA was extracted using a solution of Phenol, Chloroform, and Isoamyl Alcohol. The EBV detection and typing were performed using nested polymerase chain reaction. The EBV was detected in 9 (5.48%) out of the 164 samples studied, including 4 (5.26%) of the 76 SCC cases and 5 (5.68%) of the 88 samples in the control group (P&gt;0.05). The EBV was positive in 2.40% of the 83 male and 8.6% of the 81 female samples (P&gt;0.05). In terms of the histological grades of the case group, 3 (3/57) and 1 (1/13) of the EBV-positive samples were well and moderately differentiated, respectively (P&gt;0.05). For EBV typing, the 9 EBV-positive samples were tested, and it was found that 2 and 7 of the cases were EBV type I and II, respectively. Results of the current study demonstrated the low frequency of EBV in Iranian patients with OSCC, with EBV type II predominating. Further studies are required to clarify the association between EBV and OSCC.

The clinical and laboratory characteristics of infectious mononucleosis caused by the Epstein-Barr virus type 1 in hospitalized children

Aim. The aim of the study was to provide clinical and laboratory characteristics of infectious mononucleosis caused by Epstein-Barr virus (EBV) genotype 1 in hospitalized children. Materials and methods. The material of the study was blood leukocytes and saliva of children aged 1-17 years, hospitalized with a diagnosis of infectious mononucleosis caused by EBV (n=67). For differential detection of EBV-1/ EBV-2, we used an optimized one-round PCR variant with electrophoretic detection of amplification products in agarose gel. Clinical symptoms and laboratory data of patients were analyzed separately and combined into groups of signs. Statistical data processing was carried out using the R programming language and the RStudio environment. Results. In all children, only one type of virus (EBV-1) was detected in blood leukocytes and saliva. The first data on clinical and laboratory signs of EBV-1 infectious mononucleosis, characterized by a typical symptom complex, have been obtained. The age criterion for dividing patients into groups of younger (1–5 years) and older (6–17 years) age, which were characterized by the most pronounced differences in the manifestations of overt EBV-1 infection, was determined. In young children, the leading was the syndrome of intoxication, in older children, signs of cytolytic syndrome were more pronounced. According to laboratory data, in the age group of 1–5 years, monocytopenia and a decrease in hemoglobin were more often observed, and in children of 6–17 years, lymphocytosis, monocytosis, elevated levels of hepatic transaminases and hemoglobin. Conclusion. For the first time, EBV typing was performed in children with infectious mononucleosis. The dominant genotype of the virus in infectious mononucleosis in children living in the territory of a large city in the European part of Russia is EBV-1. For the first time, the characteristics of EBV-1 infection are given. The data obtained on the different severity of clinical and laboratory signs of the disease are prerequisites for continuing research aimed at finding the relationship between the characteristics of the clinical course of the infection and the genetic heterogeneity of the EBV population.

Infection with human cytomegalovirus, Epstein-Barr virus, and high-risk types 16 and 18 of human papillomavirus in EGFR-mutated lung adenocarcinoma

To assess the frequency of human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), and high-risk types of human papillomavirus (HPV16 and HPV18) infections in lung adenocarcinoma samples.Lung adenocarcinoma cytological smears and their DNA isolates were obtained from patients hospitalized at the Department for Lung Diseases Jordanovac, Zagreb, in 2016 and 2017. Overall, 67 lung adenocarcinoma samples were examined: 34 with epidermal growth factor receptor gene (EGFR) mutations and 33 without EGFR mutations. The EGFR mutation status and virus presence were assessed with a polymerase chain reaction, and random samples were additionally tested for EBV with Sanger sequencing. HCMV, EBV, HPV16, and HPV18 infections were evaluated in relation to EGFR mutation, smoking status, and sex. A meta-analysis of available data about HPV infection in non-small cell lung cancer was performed.More frequent HCMV, EBV, HPV16, and HPV18 infections were observed in lung adenocarcinoma samples with EGFR mutations than in samples without these mutations. Coinfection of the investigated viruses was observed only in lung adenocarcinoma samples with mutated EGFR. In the group with EGFR mutations, smoking was significantly associated with HPV16 infection. The meta-analysis showed that non-small cell lung cancer patients with EGFR mutations had a higher odds of HPV infection.HCMV, EBV, and high-risk HPV infections are more frequent in EGFR-mutated lung adenocarcinomas, which indicates a possible viral impact on the etiology of this lung cancer subtype.