Epstein-Barr Virus Expression Research Articles

SOX11 expression is restricted to EBV-negative Burkitt lymphoma and is associated with molecular genetic features

AbstractSRY-related HMG-box gene 11 (SOX11) is a transcription factor overexpressed in mantle cell lymphoma (MCL), a subset of Burkitt lymphomas (BL) and precursor lymphoid cell neoplasms but is absent in normal B cells and other B-cell lymphomas. SOX11 has an oncogenic role in MCL but its contribution to BL pathogenesis remains uncertain. Here, we observed that the presence of Epstein-Barr virus (EBV) and SOX11 expression were mutually exclusive in BL. SOX11 expression in EBV-negative (EVB-) BL was associated with an IG∷MYC translocation generated by aberrant class switch recombination, whereas in EBV-negative (EBV−)/SOX11-negative (SOX11−) tumors the IG∷MYC translocation was mediated by mistaken somatic hypermutations. Interestingly, EBV− SOX11-expressing BL showed higher frequency of SMARCA4 and ID3 mutations than EBV−/SOX11− cases. By RNA sequencing, we identified a SOX11–associated gene expression profile, with functional annotations showing partial overlap with the SOX11 transcriptional program of MCL. Contrary to MCL, no differences on cell migration or B-cell receptor signaling were found between SOX11− and SOX11-positive (SOX11+) BL cells. However, SOX11+ BL showed higher adhesion to vascular cell adhesion molecule 1 (VCAM-1) than SOX11− BL cell lines. Here, we demonstrate that EBV− BL comprises 2 subsets of cases based on SOX11 expression. The mutual exclusion of SOX11 and EBV, and the association of SOX11 with a specific genetic landscape suggest a role of SOX11 in the early pathogenesis of BL.

Epstein-Barr Virus-Associated Lymphomatoid Papules: A Sign of Immunosuppression Resembling Lymphomatoid Papulosis.

Epstein-Barr virus (EBV)-positive lymphoproliferative disorders associated with immunodeficiency constitute a spectrum of lymphoid and plasma cell proliferations that vary in cytomorphology, immunophenotype, and clinical behavior. CD30-positive cutaneous lymphocytic infiltrates with EBV expression and lymphomatoid papulosis-like presentations have been rarely reported. This retrospective study assessed the clinical and histopathological characteristics of EBV-positive cases with papulonodular morphologies and CD30 positivity seen by Northwestern Medicine Dermatopathology. Twelve patients (7M:5F, mean age 69 years) were presented with papular cutaneous lesions without antecedent patch/plaque disease. Nine cases were associated with known immunosuppression in the setting of transplant-related therapies (n = 4), hematopoietic malignancy (n = 2), post-transplant hematopoietic malignancy (n = 1), and autoimmune disease treatment (n = 2). Two patients had age-related immunosenescence. Four patients demonstrated EBV viremia; for 2 patients, this finding comprised the first sign of immunosuppression. Workup was negative for systemic lymphoma in all patients. Various treatment strategies were used, including observation (n = 3), discontinuation/reduction of immunosuppression (n = 3), rituximab (n = 4), and steroids (n = 4). At mean 30-month follow-up, 4 patients (33.3%) were alive, 3 with and 1 without disease. Eight patients (67.6%) had died, 3 after lesional resolution and 5 with recurrent disease. Biopsies revealed mixed lymphoid infiltrates composed of atypical CD30-positive T cells (n = 5) or B cells (n = 7) with variable EBV-encoded small RNA expression. These cases suggest clinicopathologic presentations resembling lymphomatoid papulosis with atypical, large CD30-positive, EBV-positive cells could comprise first sign of potentially serious immunodeficiency and should prompt evaluation for EBV viremia. These cases also broaden the current picture of immunodeficiency-associated lymphoproliferative disorders to include lymphomatoid papulosis-like clinical presentations.

Viral-induced inflammation can lead to adverse pregnancy outcomes.

Parvoviruses are DNA viruses of small size. There have been a number of reports indicating the possible effects of B19 infections during pregnancy. These effects include spontaneous abortions, stillbirth, fetal damage, and quite often, fetal anemia with hydrops fetalis.

Epstein-Barr virus infectious particles initiate B cell transformation and modulate cytokine response.

The Epstein-Barr virus efficiently infects and transforms B lymphocytes. During this process, infectious viral particles transport the viral genome to the nucleus of target cells. We show here that these complex viral structures serve additional crucial roles by activating transcription of the transforming genes encoded by the virus. We show that components of the infectious particle sequentially activate proinflammatory B lymphocyte signaling pathways that, in turn, activate viral gene expression but also cause cytokine release. However, virus infection activates expression of ZFP36L1, an RNA-binding stress protein that limits the length and the intensity of the cytokine response. Thus, the infectious particles can activate viral gene expression and initiate cellular transformation at the price of a limited immune response.

Epstein-Barr virus predominance and immunological abnormalities in oral squamous cell carcinoma

BACKGROUND: Both benign and harmful head and neck disorders have been associated with the Epstein-Barr virus (EBV). Many studies have connected EBV to oral squamous cell carcinoma (OSCC). Oral squamous cell carcinoma is the most prevalent type of cancer. Fresh tissue samples from patients with OSCC were tested for the presence of Epstein-Barr virus. OBJECTIVE: To determine the frequency of EBV and IL10 expression in OSCC patients. MATERIALS AND METHODS: Fifty individuals with OSCC and 25 with clinically healthy oral mucosa were studied. In situ hybridization was used for the detection of EBV. Serum IL-10 levels were also evaluated in patients and controls using a human IL-10 ELISA Kit. RESULTS: EBV was detected in 4 healthy patients, 6 with moderately differentiated OSCC, 10 with poorly differentiated OSCC, and 19 with undifferentiated OSCC. These differences were statistically significant (p<0.05). IL-10 expression was more common in OSCC diagnostic groups than healthy controls, and the difference in blood IL-10 levels between patients and controls was statistically significant (p<0.01). CONCLUSION: The prevalence of EBV in OSCC suggests its possible role in oral cavity malignancy. On the other hand, IL-10 is expressed at higher levels in OSCC biopsies; such elevated concentrations may promote viral spread.

Positive correlation between latent Epstein-Barr virus infection and severity of illness in inflammatory bowel disease patients.

Emerging studies indicate the critical involvement of microorganisms, such as Epstein-Barr virus (EBV), in the pathogenesis of inflammatory bowel disease (IBD). Immunosuppressive therapies for IBD can reactivate latent EBV, complicating the clinical course of IBD. Moreover, the clinical significance of EBV expression in B lymphocytes derived from IBD patients' intestinal tissues has not been explored in detail. To explore the clinical significance of latent EBV infection in IBD patients. Latent EBV infection was determined by double staining for EBV encoded RNA and CD20 in colon specimens of 43 IBD patients who underwent bowel resection. Based on the staining results, the patients were divided into two groups, according to their latent EBV infection states - negative (n = 33) and positive (n = 10). Illness severity of IBD were assigned according to Crohn's disease activity index (ulcerative colitis) and Mayo staging system (Crohn's disease). The clinic-pathological data were analyzed between the two different latent EBV groups and also between the mild-to-moderate and severe disease groups. Systolic pressure (P = 0.005), variety of disease (P = 0.005), the severity of illness (P = 0.002), and pre-op corticosteroids (P = 0.025) were significantly different between the EBV-negative and EBV-positive groups. Systolic pressure (P = 0.001), variety of disease (P = 0.000), pre-op corticosteroids (P = 0.011) and EBV infection (P = 0.003) were significantly different between the mild-to-moderate and severe disease groups. IBD patients with latent EBV infection may manifest more severe illnesses. It is suggested that the role of EBV in IBD development should be further investigated, latent EBV infection in patients with serious IBD should be closely monitored, and therapeutic course should be optimized.

Oncogenic role of LMP1 expression in Epstein-Barr virus associated with nasopharyngeal carcinoma: A literature review

Oncogenic role of LMP1 expression in Epstein-Barr virus associated with nasopharyngeal carcinoma: A literature review

Oncogenic role of LMP1 expression in Epstein-Barr virus associated with nasopharyngeal carcinoma: A literature review

Nasopharyngeal carcinoma (NPC) is a malignant carcinoma that appears in the epithelial lining of the nasopharyngeal mucosa. Genetic predisposition, environmental risk factors, nutritional risk factors, and Epstein-Barr virus (EBV) infection are the main risk factors for NPC. Epstein-Barr virus, a human cancer-associated virus, is present in more than 90% of people worldwide. Latent Membrane Protein 1 (LMP1) has been identified as the main Epstein-Barr virus converting oncoprotein. LMP1 was shown to be present in NPC tumor tissues, suggesting a potential role for LMP1 in EBV-mediated carcinogenesis. LMP1 exerts several oncogenic properties and has the ability to transform epithelial cells through the activation of numerous signaling pathways and the control of the expression of different oncogenes and tumor-suppressor genes. the process by which tumor traits such cell proliferation, apoptosis resistance, invasion and motility, and angiogenesis are produced and maintained.

Reshaping tumor immune microenvironment by Epstein-Barr virus activation in the stroma of colorectal cancer

The formation of tumor immune microenvironment (TIM) is complicated and poorly understood. Little is known about the effect of a viral infection potentially inducing an additional immune response in the TIM. Here, we identify Epstein-Barr virus (EBV) expression in the TIM in colorectal cancer (CRC) tissue through EBV-encoded RNA in-situ hybridization and RNA sequencing data and investigate the effects of EBV on TIM composition and clinical outcomes. EBV was detected in tumor-infiltrating lymphocytes, but not in cancer cells. EBV positivity was associated with older age, male sex, and SMAD4 mutations. EBV-positive tumors were characterized by enrichment in chemokine/cytokine signaling pathways and altered immune cell composition, including plasma and CD4 Tcells, as well as cancer cells intrinsically enriched pathways related to immune tolerance, leading to poor prognosis. In conclusion, we identified EBV expression in TIM and suggested its association with poor prognosis by altering the TIM in CRC.

Identifying the key genes of Epstein-Barr virus-regulated tumour immune microenvironment of gastric carcinomas.

The Epstein-Barr virus (EBV) is involved in the carcinogenesis of gastric cancer (GC) upon infection of normal cell and induces a highly variable composition of the tumour microenvironment (TME). However, systematic bioinformatics analysis of key genes associated with EBV regulation of immune infiltration is still lacking. In the present study, the TCGA and GEO databases were recruited to analyse the association between EBV infection and the profile of immune infiltration in GC. The weighted gene co-expression analysis (WGCNA) was applied to shed light on the key gene modules associated with EBV-associated immune infiltration in GC. 204 GC tissues were used to analysed the expression of key hub genes by using the immunohistochemical method. Real-time PCR was used to evaluate the association between the expression of EBV latent/lytic genes and key immune infiltration genes. Our results suggested that EBV infection changed the TME of GC mainly regulates the TIICs. The top three hub genes of blue (GBP1, IRF1, and LAP3) and brown (BIN2, ITGAL, and LILRB1) modules as representative genes were associated with EBV infection and GC immune infiltration. Furthermore, EBV-encoded LMP1 expression is account for the overexpression of GBP1 and IRF1. EBV infection significantly changes the TME of GC, and the activation of key immune genes was more dependent on the invasiveness of the whole EBV virion instead of single EBV latent/lytic gene expression.

ABCL-121 Immunohistochemistry Expression of MICA, MICB, and Epstein-Barr Virus (EBV) in Lymph Node Biopsy of Patients With De Novo Non-Hodgkin Lymphoma

We recently found an increase soluble expression of MICA and MICB in patients with de novo non-Hodgkin lymphoma, negatively affecting the patient's immune response. Now we want to evaluate the expression of MIC-A and MIC-B ligands directly at the site of injury. On the other hand, EBV is the etiological agent most related to the development of cancers, mainly lymphomas, however, in our country there are still no data that evaluate the presence of EBV infection in neoplastic cells of patients with non-Hodgkin lymphoma and its possible relationship with the expression of MICA and MICB. To determine the expression of MICA and MICB in the lymph node and whether Epstein Barr virus infection in patients with de novo non-Hodgkin lymphoma favors the soluble expression of MIC-A and MIC-B. Sections will be obtained from the paraffin-embedded block of the neoplastic lymph node of each patient to determine the presence of EBV by LMP-1 expression, as well as the expression of MIC-A, MIC-B by immunohistochemistry. 30 paraffin blocks from patients diagnosed with non-Hodgkin lymphoma were worked on. Of the 30 blocks analyzed, 16 were DLBCL, 6 FL, 5 mycosis fungoides, 1 MCL, 1 NLTCL, and 1 ALCL. The expression of the LMP-1 protein was found in 57%. All cases of mycosis fungoides were positive for EBV and all cases of follicular lymphoma were negative. In all patients, the staining was positive for the presence of MIC-A as well as for MIC-B. The staining pattern for MIC-A was stronger and more massive than for MIC-B, which could indicate a greater expression of MIC-A than MIC-B in our patients, which is consistent with what was reported in the serology of these patients in our previous work.

The Prevalence and the Patterns of the Expression of Latent Epstein-Barr Virus in Hodgkin’s and Non-Hodgkin’s Lymphomas Among Patients in Oman: Immunohistochemistry Versus in Situ Hybridization.

Epstein Barr virus (EBV) has been incriminated in the pathogenesis of Hodgkin’s lymphoma (HL) and non-Hodgkin’s lymphoma (NHL). The present study aimed to investigate the prevalence and the pattern of the expression of Epstein-Barr in HL and NHL tissue samples obtained from Omani patients attending Sultan Qaboos University Hospital (SQUH). Besides, to compare the sensitivity and specificity of immunohistochemistry(IHC) and in situ hybridization (ISH) for the detection of EBV in HL and NHL and finally to have more understanding of the pathogenesis of EBV in HL and NHL among patients in Oman. Formalin-fixed paraffin-embedded tissue samples consisting of 26 Hodgkin and 34 non-Hodgkin lymphomas were assessed for the presence of EBV by IHC to detect Latent membrane protein (LMP), expression and by using ISH to detect Epstein -Barr encoded RNAs (EBERs). The expression of LMP and EBERs were detected respectively in 46.2% and 57.7% of Hodgkin’s lymphoma cases and were detected in 11.8% and 14.7% respectively of non-Hodgkin’s lymphoma cases. The intensity of LMP-1 and EBER expression was significantly high in mixed cellularity compared to other subtypes. The expression of EBV was detected in transformed cells in both HL & NHL. The expression of EBV in transformed cells in both HL and NHL indicates that EBV may play a pro vital role in the pathogenesis of HL and NHL among patients in Oman. Moreover, this study indicates that IHC is to some degree compatible in terms of sensitivity and specificity to ISH in the detection of EBV in HL and NHL.

Relative Expression of mRNA BARF1 Epstein-Barr Virus from Tumor Tissue Biopsy in Formalin-Fixed Paraffin-Embedded in Nasopharyngeal Carcinoma

Nasopharyngeal carcinoma (NPC) is a malignant tumor originating from nasopharyngeal epithelial cells. NPC is endemic in some regions, especially in Southeast Asia countries. NPC is also a multifactorial disease involving environmental factors, genetic factors, and infection from Epstein-Barr virus (EBV). According to WHO classification, Undifferentiated NPC is histopathologically associated with EBV infection and categorized into WHO subtype 3. EBV has two phases in its infection cycle: the lytic and latent phases. The BARF1 gene is a mediator of the transition from the latent phase to the lytic phase. Previous studies suggest measurement of EBV mRNA activity at the primary tumor site in the nasopharyngeal reflects the pathogenesis of NPC rather than measuring circulating EBV DNA or serological diagnosis. This study aimed to determine the relative expression potential of BARF1 mRNA at different tumor stages in NPC patients as a predictor of NPC pathogenesis. This research design was a descriptive research method in the form of a cross-sectional study. The samples used were 22 patients diagnosed as NPC WHO class III at the Anatomical Pathology Section of Prof. Dr. Margono Soekarjo, Purwokerto, who met the inclusion criteria. The relative expression of BARF1 mRNA was carried out using the one-step real-time RT-PCR technique and then calculated using a formula of 2-ΔCt. The T-test was used to compare the relative expression of early and late-stage BARF1 mRNA. The relative expression of BARF1 mRNA in the late-stage advanced stage (n = 6; 0.708292-0.840177; med = 0.7164655) was increased compared to the early stage (n = 2; 0.708841-0.712423; med = 0. 710632).

Major Role for Cellular MicroRNAs, Long Noncoding RNAs (lncRNAs), and the Epstein-Barr Virus-Encoded BART lncRNA during Tumor Growth In Vivo.

ABSTRACTThis study assessed the effects of Epstein-Barr virus (EBV) and one form of virally encoded BART long noncoding RNAs (lncRNAs) on cellular expression in epithelial cells grown in vitro and as tumors in vivo determined by high-throughput RNA sequencing of mRNA and small RNAs. Hierarchical clustering based on gene expression distinguished the cell lines from the tumors and distinguished the EBV-positive tumors and the BART tumors from the EBV-negative tumors. EBV and BART expression also induced specific expression changes in cellular microRNAs (miRs) and lncRNAs. Multiple known and predicted targets of the viral miRs, the induced cellular miRs, and lncRNAs were identified in the altered gene set. The changes in expression in vivo indicated that the suppression of growth pathways in vivo reflects increased expression of cellular miRs in all tumors. In the EBV and BART tumors, many of the targets of the induced miRs were not changed and the seed sequences of the nonfunctional miRs were found to have homologous regions within the BART lncRNA. The inhibition of these miR effects on known targets suggests that these induced miRs have reduced function due to sponging by the BART lncRNA. This composite analysis identified the effects of EBV on cellular miRs and lncRNAs with a functional readout through identification of the simultaneous effects on gene expression. Major shifts in gene expression in vivo are likely mediated by effects on cellular noncoding RNAs. Additionally, a predicted property of the BART lncRNA is to functionally inhibit the induced cellular miRs.

Expression of the Chemokine Receptor CCR1 in Burkitt Lymphoma Cell Lines Is Linked to the CD10-Negative Cell Phenotype and Co-Expression of the EBV Latent Genes EBNA2, LMP1, and LMP2.

Chemokines and their receptors regulate the migration of immune cells and the dissemination of cancer cells. CCR1, CCR2, CCR3, and CCR5 all belong to a single protein homology cluster and respond to the same inflammatory chemokines. We previously reported that CCR1 and CCR2B are induced upon Epstein-Barr virus (EBV) infection of B cells in vitro. EBV is present in almost all cases of endemic Burkitt lymphoma (BL); however, the contribution of EBV in the pathogenesis of the disease is not fully understood. Here, we analyzed the relation of the expression of CCR1, CCR2, CCR3, and CCR5, the EBV DNA load and expression of EBV latent genes in nine EBV-carrying and four EBV-negative BL cell lines. We revealed that CCR1 is expressed at high mRNA and protein levels in two CD10-negative BL cell lines with co-expression of the EBV latent genes EBNA2, LMP1, and LMP2. Low levels of CCR2 transcripts were found in three BL cell lines. CCR3 and CCR5 transcripts were hardly detectable. Our data suggest that in vivo, CCR1 may be involved in the dissemination of BL cells and in the selection of BL cells with restricted EBV gene expression programs.

Herpesviruses and their genetic diversity in the blood virome of healthy individuals: effect of aging

BackgroundAs we age, the functioning of the human immune system declines. The results of this are increases in morbidity and mortality associated with infectious diseases, cancer, cardiovascular disease, and neurodegenerative disease in elderly individuals, as well as a weakened vaccination response. The aging of the immune system is thought to affect and be affected by the human virome, the collection of all viruses present in an individual. Persistent viral infections, such as those caused by certain herpesviruses, can be present in an individual for long periods of time without any overt pathology, yet are associated with disease in states of compromised immune function. To better understand the effects on human health of such persistent viral infections, we must first understand how the human virome changes with age. We have now analyzed the composition of the whole blood virome of 317 individuals, 21–70 years old, using a metatranscriptomic approach. Use of RNA sequencing data allows for the unbiased detection of RNA viruses and active DNA viruses.ResultsThe data obtained showed that Epstein-Barr virus (EBV) was the most frequently expressed virus, with other detected viruses being herpes simplex virus 1, human cytomegalovirus, torque teno viruses, and papillomaviruses. Of the 317 studied blood samples, 68 (21%) had EBV expression, whereas the other detected viruses were only detected in at most 6 samples (2%). We therefore focused on EBV in our further analyses. Frequency of EBV detection, relative EBV RNA abundance and the genetic diversity of EBV was not significantly different between age groups (21–59 and 60–70 years old). No significant correlation was seen between EBV RNA abundance and age. Deconvolution analysis revealed a significant difference in proportions of activated dendritic cells, macrophages M1, and activated mast cells between EBV expression positive and negative individuals.ConclusionsAs it is likely that the EBV RNA quantified in this work is derived from reactivation of the latent EBV virus, these data suggest that age does not affect the rate of reactivation nor the genetic landscape of EBV. These findings offer new insight on the genetic diversity of a persistent EBV infection in the long-term.

Evaluation of Epstein–Barr virus expression in oral squamous cell carcinomas

Oral squamous cell carcinoma (OSCC) is one of the most common cancers. Epstein-Barr virus (EBV) has been related to throat-esophageal and gastric cancers. The aim of this study was to evaluate the prevalence of EBV in OSCC. This descriptive-analytical cross-sectional study was performed on 48 samples recorded in the archives of the Oral Pathology Department of Isfahan Dental School with definitive diagnosis of OSCCs prepared by excisional biopsy. Samples were selected in different age groups, locations, and genders. The grade of the tumor malignancy was determined based on Annreroth's classification. The EBV expression was determined by immunohistochemical (IHC) staining. The data were entered into SPSS software and statistically analyzed by t-test, Chi-square, and Fisher's exact test. Significance level was considered P < 0.05. IHC staining for EBV was positive in 25 samples (52%). There was no significant relationship between EBV expression and mean age, gender, clinical feature, and grade of tumor differentiation (P > 0.05). A significant difference was observed between the EBV expression and location (P = 0.035). Furthermore, a significant difference was observed between the grade of tumor and staining intensity distribution index of EBV (P = 0.005). EBV expression was observed in most of the OSCCs, especially in poorly differentiated tumors. The pathogenesis of OSCCs may be related with EBV. OSCCs in buccal mucosa and floor of the mouth have more frequently of EBV expression. Future studies on the mechanisms of EBV and their role in OSCC are required with larger sample sizes.

Prognostic Role of the Expression of Latent-Membrane Protein 1 of Epstein-Barr Virus in Classical Hodgkin Lymphoma.

The prognostic impact of the presence of Epstein–Barr virus (EBV) in classical Hodgkin lymphoma (cHL) is controversial. Previous studies reported heterogeneous results, rendering difficult the clinical validation of EBV as a prognostic biomarker in this lymphoma. The objective of this study was to evaluate the survival impact of the expression of EBV Latent-Membrane Protein 1 (EBV-LMP1) in tumoral Hodgkin–Reed–Sternberg (HRS) cells of primary diagnostic samples of cHL. Formalin-Fixed Paraffin-Embedded (FFPE) lymph node samples from 88 patients with cHL were analyzed. Patients were treated with the standard first-line chemotherapy (CT) with Adriamycin, Bleomycin, Vinblastine and Dacarbazine (ABVD) followed by radiotherapy. The Kaplan–Meier method and the Cox proportional hazards model were used for carrying out the survival analysis. In order to investigate whether the influence of EBV was age-dependent, analyses were performed both for patients of all ages and for age-stratified subgroups. In bivariate analysis, the expression of EBV was associated with older age (p = 0.011), mixed cellularity subtype cHL (p < 0.001) and high risk International Prognostic Score (IPS) (p = 0.023). Overall survival (OS) and progression-free survival (PFS) were associated with the presence of bulky disease (p = 0.009) and advanced disease at diagnosis (p = 0.016). EBV-positive cases did not present a significantly lower OS and PFS in comparison with EBV-negative cases, for all ages and when stratifying for age. When adjusted for covariates, absence of bulky disease at diagnosis (HR: 0.102, 95% CI: 0.02–0.48, p = 0.004) and limited disease stages (I–II) (HR: 0.074, 95% CI: 0.01–0.47, p = 0.006) were associated with a significant better OS. For PFS, limited-disease stages also retained prognostic impact in the multivariate Cox regression (HR: 0.145, 95% CI: 0.04–0.57, p = 0.006). These results are of importance as the early identification of prognostic biomarkers in cHL is critical for guiding and personalizing therapeutic decisions. The prognostic role of EBV in cHL could be modulated by the type of CT protocol employed and interact with the rest of presenting features.

HLA Expression in Relation to HLA Type in Classic Hodgkin Lymphoma Patients.

Simple SummaryClassic Hodgkin lymphoma (cHL) is a B-cell malignancy with involvement of Epstein–Barr virus (EBV) in about 30% of the European population. The risk to develop cHL is strongly linked to genetic variants in the human leukocyte antigen (HLA) genomic region and to certain HLA alleles. This may be caused by the function of HLA alleles, or by genetic linkage to non-HLA genes. HLA can present EBV-derived and tumour-cell specific antigens and this may lead to anti-tumour immune responses. However, the tumour cells downregulate HLA expression in a proportion of the cases, which may result in immune escape. In this study, we tested whether the loss of HLA expression is related to the presence of certain protective HLA alleles. We found that loss and retention of HLA expression is indeed associated with presence of known susceptibility HLA alleles. These findings suggest that HLA itself is involved in development of cHL.Several human leukocyte antigen (HLA) alleles are strongly associated with susceptibility to classic Hodgkin lymphoma (cHL), also in subgroups stratified for presence of the Epstein–Barr virus (EBV). We tested the hypothesis that the pressure on cHL tumour cells to lose HLA expression is associated with HLA susceptibility alleles. A meta-analysis was carried out to identify consistent protective and risk HLA alleles in a combined cohort of 839 cHL patients from the Netherlands and the United Kingdom. Tumour cell HLA expression was studied in 338 cHL cases from these two cohorts and correlated to the presence of specific susceptibility HLA alleles. Carriers of the HLA-DRB1*07 protective allele frequently lost HLA class II expression in cHL overall. Patients carrying the HLA-DRB1*15/16 (DR2) risk allele retained HLA class II expression in EBV− cHL and patients with the HLA-B*37 risk allele retained HLA class I expression more frequently than non-carriers in EBV+ cHL. The other susceptibility alleles showed no significant differences in expression. Thus, HLA expression by tumour cells is associated with a subset of the protective and risk alleles. This strongly suggests that HLA associations in cHL are related to peptide binding capacities of specific HLA alleles.

Programmed death ligand 1 expression and CD8+T lymphocyte infiltration in salivary gland lymphoepithelial carcinoma

Objective: To study the expression of programmed death ligand-1 (PD-L1) in tumor cells and CD8+T lymphocytes in tumor infiltrating lymphocytes, and to analyze the correlation of PD-L1 expression with infiltration of CD8+T lymphocytes and clinicopathologic features in salivary gland lymphoepithelial carcinoma (LEC). Methods: Forty-two cases of primary salivary LECs and 21 cases of secondary salivary LECs were enrolled at the Department of Oral Pathology, Shanghai Ninth People's Hospital, Shanghai Jiaotong University between 2015 and 2017. The expression of Epstein-Barr (EB) virus, PD-L1 and CD8 was examined using chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) staining. The data were analyzed using SPSS 23.0 software package. Results: EB virus was detected in 61 cases (61/63, 96.8%), including 42 (42/42, 100%) primary LECs and 19 (19/21, 90.5%) secondary LECs. The PD-L1 positive rate (score ≥1) was 97.6% (41/42), and its high-expression rate (score ≥20) was 78.6% (33/42) in primary LECs. The PD-L1 positive rate (score ≥1) was 71.4% (15/21), and its high-expression rate (≥20) was 38.1% (8/21) in secondary LECs. However, the PD-L1 positive rate (score ≥1, P=0.004) and high-expression rate (score ≥20, P=0.001) in primary LECs were higher than those in secondary LECs. There was no difference in the infiltration degree of CD8+T lymphocytes between primary and secondary LECs. There was a significant correlation between the expression of PD-L1 and CD8 in primary LECs (P=0.001) and in secondary LECs (P=0.048), respectively. Conclusions: There is PD-L1 expression in primary and secondary salivary LECs, while the expression rate is higher in primary LECs than secondary LECs. The combination of PD-L1 expression and CD8+T lymphocytes' presence suggest that most LEC patients might be responsive to immunotherapy, and primary LECs might be more significantly responsive than secondary LECs.