Tumor Epstein-Barr Virus Status Research Articles

Genetic subgroups inform on pathobiology in adult and pediatric Burkitt lymphoma

AbstractBurkitt lymphoma (BL) accounts for most pediatric non-Hodgkin lymphomas, being less common but significantly more lethal when diagnosed in adults. Much of the knowledge of the genetics of BL thus far has originated from the study of pediatric BL (pBL), leaving its relationship to adult BL (aBL) and other adult lymphomas not fully explored. We sought to more thoroughly identify the somatic changes that underlie lymphomagenesis in aBL and any molecular features that associate with clinical disparities within and between pBL and aBL. Through comprehensive whole-genome sequencing of 230 BL and 295 diffuse large B-cell lymphoma (DLBCL) tumors, we identified additional significantly mutated genes, including more genetic features that associate with tumor Epstein-Barr virus status, and unraveled new distinct subgroupings within BL and DLBCL with 3 predominantly comprising BLs: DGG-BL (DDX3X, GNA13, and GNAI2), IC-BL (ID3 and CCND3), and Q53-BL (quiet TP53). Each BL subgroup is characterized by combinations of common driver and noncoding mutations caused by aberrant somatic hypermutation. The largest subgroups of BL cases, IC-BL and DGG-BL, are further characterized by distinct biological and gene expression differences. IC-BL and DGG-BL and their prototypical genetic features (ID3 and TP53) had significant associations with patient outcomes that were different among aBL and pBL cohorts. These findings highlight shared pathogenesis between aBL and pBL, and establish genetic subtypes within BL that serve to delineate tumors with distinct molecular features, providing a new framework for epidemiologic, diagnostic, and therapeutic strategies.

ABCL-542 Prevalence and Characterization of EBV-Negative Post-Transplant Lymphoproliferative Disorder

A common and potentially fatal post-solid organ transplant (SOT) complication is post-transplant lymphoproliferative disorder (PTLD). While the association of Epstein-Barr virus (EBV) with the development of PTLD is well recognized, little is known about the etiology of EBV-negative PTLD. To characterize the differences between EBV-positive and EBV-negative PTLD to better understand the pathogenesis of EBV-negative PTLD. This was a retrospective study that reviewed patients who developed PTLD after SOT. Data was collected on PTLD patients (>18 years) from 2011 to 2021. Data included demographics, SOT type, time from SOT to PTLD development, tumor EBV status, PTLD site, histopathological morphology, tumor markers, and overall survival. EBV tumor status was compared using χ2, Fisher's Exact, and Wilcoxon Rank Sum tests. A single, large academic center. A total of 99 patients met the inclusion criteria. Of those, 45.5% were EBV-positive and 54.5% were EBV-negative. Tumor EBV status did not differ by race, but males were more likely to be EBV-negative (63% v. 44.4% of females, p=0.065). EBV tumor status did not differ by SOT type. The interval between SOT and PTLD diagnosis was longer in EBV-negative patients (8.68 years) than EBV-positive patients (4.93 years) (p=0.001). There was a significant difference in EBV-negative PTLD site (p=0.046) and PTLD subtypes (p=0.002). Site of EBV-negative PTLD consisted of gastrointestinal (38.9%), head/neck (11.1%), liver/spleen (11.1%), CNS (3.7%), other lymph (24%), and other non-lymph (11.1%). Monomorphic (65.5%) and T-cell (90%) subtypes were more common in EBV-negative patients, compared to the polymorphic (15.4%) and plasmablastic/plasmacytic (27.3%) subtypes. Expression of CD20 did not differ by tumor EBV status. Those who were CD30-negative were more likely to be EBV-negative (90% v. 25%, p=<0.001). Those who were CD138-negative were more likely to be EBV-negative (75% v. 26.7%, p=0.012). There was no difference in overall survival by tumor EBV status. This single institution retrospective review shows that there are distinct differences between EBV-negative and EBV-positive PTLD. EBV-negative PTLD has a longer interval from SOT to PTLD development, is more likely to occur in the gastrointestinal tract, and is more likely to have a monomorphic morphology.

Identification of anti-Epstein-Barr virus (EBV) antibody signature in EBV-associated gastric carcinoma.

Around 10% of gastric carcinomas (GC) contain Epstein-Barr virus (EBV) DNA. We characterized the GC-specific antibody response to this common infection, which may provide a noninvasive method to detect EBV-positive GC and elucidate its contribution to carcinogenesis. Plasma samples from EBV-positive (n = 28) and EBV-negative (n = 34) Latvian GC patients were immune-profiled against 85 EBV proteins on a multi-microbial Nucleic Acid Programmable Protein Array (EBV-NAPPA). Antibody responses were normalized for each sample as ratios to the median signal intensity (MNI) across all antigens, with seropositivity defined as MNI ≥ 2. Antibodies with ≥ 20% sensitivity at 95% specificity for tumor EBV status were verified by enzyme-linked immunosorbent assay (ELISA) and validated in independent samples from Korea and Poland (n = 24 EBV-positive, n = 65 EBV-negative). Forty anti-EBV IgG and eight IgA antibodies were detected by EBV-NAPPA in ≥ 10% of EBV-positive or EBV-negative GC patients, of which nine IgG antibodies were discriminative for tumor EBV status. Eight of these nine were verified and seven were validated by ELISA: anti-LF2 (odds ratio = 110.0), anti-BORF2 (54.2), anti-BALF2 (44.1), anti-BaRF1 (26.7), anti-BXLF1 (12.8), anti-BRLF1 (8.3), and anti-BLLF3 (5.4). The top three had areas under receiver operating characteristics curves of 0.81-0.85 for distinguishing tumor EBV status. The EBV-associated GC-specific humoral response was exclusively directed against lytic cycle immediate-early and early antigens, unlike other EBV-associated malignancies such as nasopharyngeal carcinoma and lymphoma where humoral response is primarily directed against late lytic antigens. Specific anti-EBV antibodies could have utility for clinical diagnosis, epidemiologic studies, and immune-based precision treatment of EBV-positive GC.

Abstract 2370: Elevated levels of anti-Epstein Barr virus (EBV) antibodies in EBV-associated gastric carcinoma

Abstract Background: Almost 10% of gastric carcinomas contain Epstein-Barr virus (EBV) DNA, which is not found in other gastrointestinal tract cancers. EBV-positive tumors have distinct clinico-epidemiologic features and are molecularly characterized by recurrent PIK3CA and absent TP53 mutations, JAK2 and PD-L1/L2 amplifications, and extreme DNA hypermethylation. Identification of the gastric cancer-specific antibody response to this common infection may elucidate its contribution to carcinogenesis and provide a non-invasive method to detect EBV-positive gastric tumors. Methods: Plasma samples from Latvian patients with EBV-positive (n=28) and -negative (n=34) gastric cancer were immune-profiled against 85 EBV proteins expressed in situ on a multi-microbiome Nucleic Acid Programmable Protein Array (NAPPA). Antibody responses were normalized as ratios to the median signal intensity across all antigens, with seropositivity defined as ratios ≥ 2. Antibodies with ≥ 20% sensitivity at 95% specificity for EBV status were verified by enzyme-linked immunosorbent assay (ELISA) and then validated in independent samples from Korea and Poland (n=24 EBV-positive, n=65 EBV-negative). Results: Forty anti-EBV IgG and eight IgA antibodies were present by NAPPA in ≥ 10% of EBV-positive or EBV-negative gastric cancer patients, of which nine were sensitive and specific for tumor EBV status. Eight of these nine were verified and seven were validated by ELISA: anti-LF2 (odds ratio=110.0, p&amp;lt;0.01), anti-BORF2 (54.2, p&amp;lt;0.01), anti-BALF2 (44.1, p&amp;lt;0.01), anti-BaRF1 (26.7, p&amp;lt;0.01), anti-BXLF1 (12.8, p=0.01), anti-BRLF1 (8.3, p&amp;lt;0.01), and anti-BLLF3 (5.4, p=0.02). The top three antibodies had areas under the receiver operating characteristics curve of 0.81-0.85 for discriminating gastric cancer EBV status. Conclusions: This is the first comprehensive immune profiling of anti-EBV response in gastric cancer. Despite the ubiquity of EBV infection in adults, seven novel anti-EBV antibodies were identified and validated to distinguish viral involvement in gastric cancer. Unlike other major EBV-associated malignancies such as nasopharyngeal carcinoma and lymphoma, the gastric cancer-specific humoral response is exclusively directed against lytic cycle immediate-early and early antigens. EBV serology could have utility for clinical diagnosis, epidemiologic studies, and precision treatment of EBV-positive gastric cancer. Citation Format: Lusheng Song, Minkyo Song, M. Constanza Camargo, Jennifer Van Duine, Stacy Williams, Yunro Chung, Kyoung-Mee Kim, Jolanta Lissowska, Sergejs Isajevs, Sung Kim, Weimin Gao, Kailash Karthikeyan, Jin Park, Marcis Leja, Jeffrey L. Cohen, Joshua LaBaer, Ji Qiu, Charles S. Rabkin. Elevated levels of anti-Epstein Barr virus (EBV) antibodies in EBV-associated gastric carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2370.

Assessment of tumor Epstein-Barr Virus status and its impact on outcomes in intermediate and high-risk childhood classic Hodgkin Lymphoma treated at a tertiary cancer center in India

Indian studies on EBV in childhood classic Hodgkin Lymphoma (cHL) have mainly analyzed the epidemiology of EBV-positive [EBV(+)HL] or negative HL [EBV(–)HL], with limited data on outcomes. We studied a large cohort of children with intermediate and high-Risk cHL for tumor EBV status and its impact on outcomes retrospectively. Of evaluable 189 patients, 84.7% had EBV(+)HL. Positive status was significantly associated with age ≤ 10 years (p < .001), males (p = .015), non-Nodular Sclerosis (NS) histology (p = .004) and inversely with bulky-mediastinal disease (p < .001). At a median follow-up of 29-months (range1–75), 3-year Event-Free Survival (EFS) for EBV(+)HL and EBV(–)HL was 93.6%(95%CI:89.8%–97.5%), 81.1%(95%CI:67.2%–97.9%), (p = .048) and Overall Survival (OS) was 94.9%(95%CI:91.6%–98.4%), 84.6%(95%CI:71.5%–100%), (p = .075) respectively. Three-year EFS was better in males (HR-0.267,95%CI:0.078–0.916, p = .036) in EBV(+)HL and in patients with serum-albumin > 3g/dL (HR-0.117,95%CI:0.019–0.705, p = .019) in EBV(–)HL. EBV is associated with most of intermediate and high-risk childhood cHL, occurs in younger male patients with non-NS histology, with reduced incidence of bulky-mediastinal disease and favorable survival in childhood cHL.

Circulating Antibodies against Epstein-Barr Virus (EBV) and p53 in EBV-Positive and -Negative Gastric Cancer.

Epstein-Barr virus (EBV)-positive gastric cancers have clinicopathologic differences from EBV-negative tumors and lack TP53 mutation. Serologic profiles may inform viral contribution to carcinogenesis. We compared humoral responses of EBV-positive (n = 67) and EBV-negative (n = 137) patients with gastric cancer from the International EBV-Gastric Cancer Consortium. Serum antibodies against four EBV proteins, nuclear (EBNA), viral capsid (VCA), early-diffuse (EA-D), and Zta replication activator (ZEBRA), and to p53 were assessed by multiplex assays. OR of antibody level tertiles (T1-T3) were adjusted by logistic regression. We also conducted a meta-analysis of reported anti-p53 seropositivity in gastric cancer. Consistent with EBV's ubiquity, 99% of patients were seropositive for anti-EBNA and 98% for anti-VCA, without difference by tumor EBV status. Seropositivity varied between patients with EBV-positive and EBV-negative tumors for anti-EA-D (97% vs. 67%, respectively, P < 0.001) and anti-ZEBRA (97% vs. 85%, respectively, P = 0.009). Adjusted ORs (vs. T1) for patients with EBV-positive versus EBV-negative tumors were significantly elevated for higher antibodies against EBNA (2.6 for T2 and 13 for T3), VCA (1.8 for T2 and 2.4 for T3), EA-D (6.0 for T2 and 44 for T3), and ZEBRA (4.6 for T2 and 12 for T3). Antibodies to p53 were inversely associated with EBV positivity (3% vs. 15%; adjusted OR = 0.16, P = 0.021). Anti-p53 prevalence from the literature was 15%. These serologic patterns suggest viral reactivation in EBV-positive cancers and identify variation of p53 seropositivity by subtype. Anti-EBV and anti-p53 antibodies are differentially associated with tumor EBV positivity. Serology may identify EBV-positive gastric cancer for targeted therapies.

Clinicopathological characteristics of Epstein-Barr virus-positive gastric cancer in Latvia.

Epstein-Barr virus (EBV)-associated gastric cancer has been proposed to be a distinct gastric cancer molecular subtype. The prognostic significance of EBV infection in gastric cancer remains unclear and needs further investigation. Our study aimed to analyze EBV-positive and EBV-negative gastric cancer patients regarding their personal and tumor-related characteristics, and compare their overall survival. Gastric cancer patients consecutively treated at the Riga East University Hospital during 2009-2016 were identified retrospectively. Tumor EBV status was determined by in-situ hybridization for EBV-encoded RNA (EBER). Information about clinicopathological characteristics was obtained from patient questionnaires, hospital records. Overall survival was ascertained through 30 July 2017. Cox proportional hazard regression models adjusted for personal and tumor-related covariates compared survival between EBV-positive and EBV-negative patients. There were a total of 302 gastric cancer patients (61% males) with mean and SD age 63.6 ± 11.5 years. EBER positivity was present in 8.6% of tumors. EBV-positive gastric cancer patients had better survival at 80 months [adjusted hazard ratio = 0.37, 95% confidence interval (CI) = 0.19-0.72] compared to EBV-negative patients. Worse survival was observed for patients with stage III (hazard ratio = 2.76, 95% CI = 1.67-4.56) and stage IV (hazard ratio = 10.02, 95% CI = 5.72-17.57) compared to stage I gastric cancer, and overlapping and unspecified subsite (hazard ratio = 1.85; 95% CI = 1.14; 3.00) compared to distal tumors. Tumor EBV positivity is a favorable prognostic factor in gastric cancer.

Viral, immunologic, and clinical features of primary effusion lymphoma

AbstractPrimary effusion lymphoma (PEL) is an aggressive HIV-associated lymphoma with a relatively poor prognosis in the era of effective HIV therapy. Kaposi sarcoma herpesvirus (KSHV) is the etiologic agent, and ∼80% of tumors are coinfected with Epstein-Barr virus (EBV). A better understanding of how KSHV-related immune dysregulation contributes to the natural history of PEL will improve outcomes. Twenty patients with PEL diagnosed between 2000 and 2013, including 19 treated with modified infusional etoposide, vincristine, and doxorubicin with cyclophosphamide and prednisone (EPOCH), were identified. We compared their clinical, virologic, and immunologic features vs 20 patients with HIV-associated diffuse large B-cell lymphoma and 19 patients with symptomatic interleukin (IL)-6 related KSHV-associated multicentric Castleman disease. Survival analyses of treated patients with PEL were then performed to identify prognostic factors and cancer-specific mortality. Compared with HIV-associated diffuse large B-cell lymphoma, PEL was associated with significant hypoalbuminemia (P < .0027), thrombocytopenia (P = .0045), and elevated IL-10 levels (P < .0001). There were no significant differences in these parameters between PEL and KSHV-associated multicentric Castleman disease. Median overall survival in treated patients with PEL was 22 months, with a plateau in survival noted after 2 years. Three-year cancer-specific survival was 47%. EBV-positive tumor status was associated with improved survival (hazard ratio, 0.27; P = .038), and elevated IL-6 level was associated with inferior survival (hazard ratio, 6.1; P = .024). Our analysis shows that IL-6 and IL-10 levels contribute to the natural history of PEL. Inflammatory cytokines and tumor EBV status are the strongest prognostic factors. Pathogenesis-directed first-line regimens are needed to improve overall survival in PEL.

Genome-wide discovery of somatic coding and noncoding mutations in pediatric endemic and sporadic Burkitt lymphoma

AbstractAlthough generally curable with intensive chemotherapy in resource-rich settings, Burkitt lymphoma (BL) remains a deadly disease in older patients and in sub-Saharan Africa. Epstein-Barr virus (EBV) positivity is a feature in more than 90% of cases in malaria-endemic regions, and up to 30% elsewhere. However, the molecular features of BL have not been comprehensively evaluated when taking into account tumor EBV status or geographic origin. Through an integrative analysis of whole-genome and transcriptome data, we show a striking genome-wide increase in aberrant somatic hypermutation in EBV-positive tumors, supporting a link between EBV and activation-induced cytidine deaminase (AICDA) activity. In addition to identifying novel candidate BL genes such as SIN3A, USP7, and CHD8, we demonstrate that EBV-positive tumors had significantly fewer driver mutations, especially among genes with roles in apoptosis. We also found immunoglobulin variable region genes that were disproportionally used to encode clonal B-cell receptors (BCRs) in the tumors. These include IGHV4-34, known to produce autoreactive antibodies, and IGKV3-20, a feature described in other B-cell malignancies but not yet in BL. Our results suggest that tumor EBV status defines a specific BL phenotype irrespective of geographic origin, with particular molecular properties and distinct pathogenic mechanisms. The novel mutation patterns identified here imply rational use of DNA-damaging chemotherapy in some patients with BL and targeted agents such as the CDK4/6 inhibitor palbociclib in others, whereas the importance of BCR signaling in BL strengthens the potential benefit of inhibitors for PI3K, Syk, and Src family kinases among these patients.

Associations of Epstein-Barr Virus-Positive Gastric Adenocarcinoma with Circulating Mediators of Inflammation and Immune Response.

Epstein-Barr virus (EBV)-positive gastric adenocarcinoma exhibits locally intense inflammation but systemic manifestations are uncertain. Our study examined whether circulating mediators of inflammation and immune response differ by tumor EBV status. From a Latvian series of 302 gastric cancer cases, we measured plasma levels of 92 immune-related proteins in the 28 patients with EBV-positive tumors and 34 patients with EBV-negative tumors. Eight markers were statistically significantly higher with tumor EBV positivity: chemokine C-C motif ligand (CCL) 20 (Odds Ratio (OR) = 3.6; p-trend = 0.001), chemokine C-X-C motif ligand 9 (OR = 3.6; p-trend = 0.003), programmed death-ligand 1 (PD-L1; OR = 3.4; p-trend = 0.004), interleukin (IL)-10 (OR = 2.4; p-trend = 0.019), CCL19 (OR = 2.3; p-trend = 0.019), CCL11 (OR = 2.2; p-trend = 0.026), IL-17A (OR = 2.0; p-trend = 0.038) and CCL8 (OR = 1.9; p-trend = 0.049). Systemic responses to EBV-positive gastric cancer are characterized by alterations in chemokines and PD-L1. Profiling of these molecules may enable non-invasive diagnosis of EBV status when tumor tissue is unavailable. Our findings provide theoretical justification for clinical evaluations of immune checkpoint therapy for EBV-positive gastric cancer.

Abstract 4246: Associations of Epstein-Barr virus (EBV)-positive gastric cancer with circulating mediators of inflammation and immune response

Abstract Epstein-Barr virus (EBV) positivity defines one of four major molecular types of gastric cancer in The Cancer Genome Atlas (TCGA). However, viral status is not routinely determined in clinical practice and tumor samples are not generally collected in epidemiologic research. Histologically, EBV-positive gastric cancer is characterized by prominent inflammatory infiltrate. In molecular analyses from TCGA, EBV-positive gastric cancer had significantly higher expression of several chemokines, chemokine receptors and programmed death-ligand 1 (PD-L1) as compared to other molecular types combined. We hypothesized that EBV tumor status may also be reflected in profiles of circulating chemokines and other markers of immune response. We have therefore evaluated pre-treatment EDTA plasma samples of gastric cancer patients from Latvia, including 28 with EBV-positive (as assessed by EBV RNA in situ hybridization) and 34 with EBV-negative tumors, frequency-matched by age, sex, anatomical subsite and Lauren histological type. We measured 92 markers using the Proseek Inflammation Panel I (Olink Proteomics, Uppsala, Sweden). We used logistic regression to model adjusted associations between tertiles of the markers and tumor EBV positivity. P&amp;lt;0.05 was considered statistically significant. Nine markers had undetectable levels in all or almost all samples. Of 83 evaluable markers, 8 were significantly higher in patients with EBV-positive tumors vs. those with EBV-negative tumors, including Chemokine C-C motif ligand (CCL) 20 (odds ratio per tertile [OR]: 3.6; p-trend=0.001), Chemokine C-X-C motif ligand 19 (OR=3.6; p-trend=0.003), PD-L1 (OR=3.4; p-trend=0.004), interleukin (IL) 10 (OR=2.4; p-trend=0.019), CCL19 (OR=2.3; p-trend=0.019), CCL11 (OR=2.2; p-trend=0.026), IL-17A (OR=2.0; p-trend=0.038) and CCL8 (OR=1.9; p-trend=0.049). These eight markers were positively correlated with the 28 pair-wise Spearman correlations ranging from 0.14 - 0.71, of which 24 were statistically significant. A model combining the top three markers, CCL20, CXCL19 and PD-L1, discriminated tumor EBV status with an area under the receiver operating characteristics curve of 0.82. These data indicate that the local alterations of specific chemokines and PDL-1 characterizing EBV-positive gastric tumors are reflected in the systemic circulation. Profiling of these molecules may enable non-invasive diagnosis of EBV-positive gastric cancer and facilitate etiologic and translational studies in large-scale prospective cohorts that lack tumor specimens. Our findings provide additional support for evaluating PD-1 blockade and other immunomodulatory therapies for patients with EBV-positive gastric cancer. Citation Format: M. Constanza Camargo, Armands Sivins, Sergejs Isajevs, Dace Rudzite, Margaret Gulley, G. Johan Offerhaus, Marcis Leja, Charles S. Rabkin. Associations of Epstein-Barr virus (EBV)-positive gastric cancer with circulating mediators of inflammation and immune response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4246. doi:10.1158/1538-7445.AM2017-4246

A Novel Risk Locus at 6p21.3 for Epstein-Barr Virus-Positive Hodgkin Lymphoma.

A proportion of the genetic variants involved in susceptibility to Hodgkin lymphoma differ by the tumor's Epstein-Barr virus (EBV) status, particularly within the MHC region. We have conducted an SNP imputation study of the MHC region, considering tumor EBV status in 1,200 classical Hodgkin lymphoma (cHL) cases and 5,726 control subjects of European origin. Notable findings were genotyped in an independent study population of 468 cHL cases and 551 controls. We identified and subsequently replicated a novel association between a common genetic variant rs6457715 and cHL. Although strongly associated with EBV-positive cHL [OR, 2.33; 95% confidence interval (CI), 1.83-2.97; P = 7 × 10(-12)], there was little evidence for association between rs6457715 and the EBV-negative subgroup of cHL (OR, 1.06; 95% CI, 0.92-1.21), indicating that this association was specific to the EBV-positive subgroup (Phet < P = 10(-8)). Furthermore, the association was limited to EBV-positive cHL subgroups within mixed cell (MCHL) and nodular sclerosis subtypes (NSHL), suggesting that the association is independent of histologic subtype of cHL. rs6457715, located near the HLA-DPB1 gene, is associated with EBV-positive cHL and suggests this region as a novel susceptibility locus for cHL. This expands the number of genetic variants that are associated with cHL and provides additional evidence for a critical and specific role of EBV in the etiology of this disease.

Anti-Helicobacter pylori Antibody Profiles in Epstein-Barr virus (EBV)-Positive and EBV-Negative Gastric Cancer.

Helicobacter pylori is the primary cause of gastric cancer, but about 9% of cases harbor Epstein-Barr virus (EBV) in the tumor cells. There is limited evidence on the possible interaction or antagonism between these infectious agents in gastric carcinogenesis. We compared H. pylori serologic profiles of EBV-positive (n = 58) and EBV-negative (n = 111) noncardia gastric cancer patients from the United States National Cancer Institute's International EBV-Gastric Cancer Consortium. EBV positivity of tumors was assessed by in situ hybridization. Serum levels of 15 antibodies to immunogenic proteins of H. pylori (Cad, CagA, Cagδ, CagM, Catalase, GroEL, HcpC, HP0231, HP0305, HpaA, HyuA, NapA, Omp, UreA, VacA) were assessed using bead-based multiplex serology. Logistic regression models were used to adjust odds ratios (OR) for country, age, sex, and year of diagnosis. Seropositivity to individual proteins ranged up to 90% overall. Antibodies to Catalase were borderline associated with tumor EBV positivity (adjusted OR = 3.15, p = .0024, Bonferroni corrected p = .036). Distributions of other antibodies did not vary by tumor EBV status. Similarity of host-response indicates the essential etiological role of H. pylori in EBV-positive gastric cancer.

Candidate gene association studies and risk of Hodgkin lymphoma: a systematic review and meta-analysis.

To evaluate the contribution of association studies of candidate polymorphisms to inherited predisposition to Hodgkin lymphoma (HL), we conducted a systematic review and meta-analysis of published case-control studies. Of the variants examined more than once in candidate gene association studies, we identified 21 studies that reported on 12 polymorphic variants in 10 genes. Data were also extracted from a published genome wide association study to allow analysis of an additional 47 variants in a further 30 genes. Promising associations were seen in nine of the variants (p < 0.05). Given that the estimated false positive report probabilities (FPRPs) for all associations are high (i.e. FPRP > 0.2), these findings should be interpreted with caution. While studies of candidate polymorphisms may be an attractive means of identifying risk factors for HL, future studies should employ sample sizes adequately powered to identify variants having only modest effects on HL risk. Furthermore, because of aetiological heterogeneity within HL, stratification of genotyping according to age, tumour Epstein-Barr virus status and histology is essential. Copyright © 2015 John Wiley & Sons, Ltd.

Epstein-Barr virus antibodies and the risk of associated malignancies: review of the literature.

Epstein-Barr virus (EBV), a ubiquitous herpes virus that infects 90% of humans by adulthood, is linked to the development of various cancers, including nasopharyngeal carcinoma, gastric cancer, Burkitt lymphoma, non-Hodgkin lymphoma (NHL), and Hodgkin lymphoma. We reviewed the literature published since 1980 regarding an association between antibodies against EBV proteins and the risk of EBV-associated malignancies. Immunoglobulin A antibody levels that are elevated before diagnosis have consistently been associated with the risk of nasopharyngeal carcinoma, and patients with Hodgkin lymphoma have significantly higher immunoglobulin G antibody levels than disease-free controls. However, the link between the immune response to EBV and other EBV-associated malignancies was less clear. Although evidence of an association between the risk of Burkitt lymphoma and immunoglobulin G antibodies was consistent for available studies, the sample sizes were limited. Evidence for a link between antibodies against EBV and risk of either gastric cancer or NHL was inconsistent. Future investigations should account for tumor EBV status because only 7%-10% of gastric tumors and select NHL subtypes are related to EBV infection. Comparing differences in the associations between the humoral immune response to EBV and disease risk across cancers may help elucidate how this ubiquitous virus contributes to distinct tumors globally.

Infectious, autoimmune and allergic diseases and risk of Hodgkin lymphoma in children and adolescents: A Children's Oncology Group study

An infectious origin for pediatric Hodgkin lymphoma (HL) has long been suspected and Epstein-Barr virus (EBV) has been implicated in a subset of cases. Increased HL incidence in children with congenital and acquired immunodeficiencies, consistent associations between autoimmune diseases and adult HL and genome-wide association and other genetic studies together suggest immune dysregulation is involved in lymphomagenesis. Here, healthy control children identified by random digit dialing were matched on sex, race/ethnicity and age to HL diagnosed in 1989-2003 at 0-14 years at Children's Oncology Group institutions. Parents of 517 cases and 784 controls completed telephone interviews, including items regarding medical histories. Tumor EBV status was determined for 355 cases. Using conditional logistic regression, we calculated odds ratios (ORs) and 95% confidence intervals (CIs) for risk of HL. Cases were more likely to have had an infection>1 year prior to HL diagnosis (OR=1.69, 95% CI: 0.98-2.91); case siblings were also more likely to have had a prior infection (OR=2.04, 95% CI: 1.01-4.14). Parental history of autoimmunity associated with increased EBV+ HL risk (OR=2.97, 95% CI: 1.34-6.58), while having a parent (OR=1.47, 95% CI: 1.01-2.14) or sibling (OR=1.62, 95% CI: 1.11-2.36) with an allergy was associated with EBV - HL. These results may indicate true increased risk for infections and increased risk with family history of autoimmune and allergic conditions that varies by tumor EBV status, or they may be attributable to inaccurate recall. In addition to employing biomarkers to confirm the role of immune-modulating conditions in pediatric HL, future studies should focus on family based designs.

Abstract 2537: Infectious, autoimmune, and allergic diseases and risk of Hodgkin lymphoma in children and adolescents: A Children's Oncology Group (COG) study.

Abstract Relatively little research has been devoted to understanding the causes of Hodgkin lymphoma (HL) in children. HL diagnosed at &amp;lt;15 years has incidence patterns and clinical characteristics distinct from HL in older adolescents and adults. Because of these differences, different underlying causes have also been postulated. Nevertheless, similar to adult HL, an infectious origin for pediatric HL has long been suspected and Epstein-Barr virus (EBV) has been implicated in a subset of cases. Results of genome-wide association studies in adult HL, consistent associations between autoimmune diseases and HL in adults, and increased HL incidence in children with congenital and acquired immunodeficiencies together suggest a role for immune dysregulation in lymphomagenesis. Here we examined associations between personal and family histories of infectious, autoimmune, and allergic diseases and HL in children and adolescents using data from the largest case-control study conducted to date. Healthy control children identified by random digit dialing were matched on sex, race/ethnicity, and date of birth to cases diagnosed in 1989-2003 at 0-14 years at North American COG institutions. Parents of 517 cases and 784 controls completed standardized 1-hour telephone interviews, including items regarding medical histories. Tumor EBV status was determined by detection of EBV-encoded RNAs via digoxigenin-based in situ hybridization. Using multivariate conditional logistic regression, we calculated odds ratios (ORs) and 95% confidence intervals (CIs) for risk of HL overall, and by tumor EBV status (EBV+, EBV-) and HL subtypes (nodular sclerosis, mixed cellularity, lymphocyte predominant) after adjusting for income, education, and birth order. Cases (OR=1.79, 95% CI: 1.04-3.08) were more likely to have had an infection &amp;gt;1 year prior to HL diagnosis, and EBV+ cases were more likely to report prior sore throats (including strep throat, scarlet fever, tonsillitis; OR=2.19, 95% CI: 1.06-4.53) and infectious mononucleosis (OR=9.10, 95% CI: 0.81-102.3). Case siblings were also more likely to have had an infection prior to HL diagnosis (OR=2.04, 95% CI: 1.01-4.14). In index children, autoimmune (OR=0.46, 95% CI: 0.16-1.38) and allergic (OR=1.22, 95% CI: 0.93-1.59) diseases identified &amp;gt;1 year before HL diagnosis were not associated with HL overall or by EBV status or subtype. There were no notable associations for autoimmune or allergic diseases in parents or siblings. These results may indicate a true increased risk for infections and lack of association with history of conditions imparting immune dysregulation, or they may be attributable to inaccurate recall. Future studies should employ biomarkers to understand differences in immune function preceding pediatric HL. Supported by NIH Grants R01 CA047473 and K05 CA157439, and Children's Cancer Research Fund, Minneapolis, MN. Citation Format: Amy M. Linabery, Erik B. Erhardt, Rachel K. Fonstad, Seymour Grufferman, Richard F. Ambinder, Greta R. Bunin, Julie A. Ross, Logan G. Spector. Infectious, autoimmune, and allergic diseases and risk of Hodgkin lymphoma in children and adolescents: A Children's Oncology Group (COG) study. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2537. doi:10.1158/1538-7445.AM2013-2537 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

Improved survival of gastric cancer with tumour Epstein–Barr virus positivity: an international pooled analysis

Background and objectiveAbout 9% of gastric carcinomas have Epstein–Barr virus (EBV) in the tumour cells, but it is unclear whether viral presence influences clinical progression. We therefore examined a large...

HLA-A*02:01 Is Associated with Inferior Overall Survival in EBV-Associated Classical Hodgkin Lymphoma

AbstractAbstract 1526 Introduction:The risk of developing Epstein-Barr Virus (EBV)-associated classical Hodgkin Lymphoma (cHL) shows an association with class I human leukocyte antigen (HLA) genes. HLA-A*01:01 is associated with increased risk of EBV-associated cHL whereas HLA-A*02:01 is associated with decreased risk. (Hjalgrim et. al, PNAS, 2010).The effects of these two alleles are independent of each other and the effect associated with each allele is additive i.e., possession of two HLA-A*01:01 alleles is associated with greater risk than one allele. HLA-A genes are involved in cytotoxic T cell (CTL) responses to exogenous antigens, and these data suggest that EBV-specific CTL responses are important in the pathogenesis of EBV-associated cHL. This study aimed to determine if HLA-A*01:01 and A*02:01 alleles are a factor in determining clinical outcome in EBV-associated cHL. Methods:Patients with cHL were selected from two previous studies: the Scotland and Newcastle Epidemiological Study of Hodgkin’s disease (SNEHD) (Jarrett et. al, Blood, 2005) and the “Investigation of the pathogenesis of Hodgkin lymphoma” (IPHL), an ongoing collection of patients principally from the West of Scotland. Both studies have ethical approval. Clinical follow-up data were obtained through clinical records, liaison with treating physicians or the Scotland and Newcastle Lymphoma Group (SNLG) database. Clinical treatment decisions were made by individual physicians. Cases were selected on the basis of availability of tumour EBV status, a suitable sample for HLA typing and clinical follow-up data and included 257 cases from the SNEHD study and 128 from the IPHL. HLA genotyping was performed to an intermediate resolution using allele-specific PCR (Gen-probe) and subsequent bead-based sequence specific oligonucleotide (SSO) assay (Luminex). Allele assignment was performed using Quicktype for Lifematch software (Gen-probe). Results were analysed for all cases, EBV-associated cases only, and EBV-negative cases only. Overall survival (OS) and Event-free survival (EFS) were analysed by number of HLA-A*01:01 alleles, number of HLA-A*02:01 alleles, and by the presence or absence of HLA-A*01:01 or HLA-A*02:01. Statistical analyses used Kaplan-Meier survival and Cox-regression analyses, adjusting for histological subtype, clinical stage, age and sex, and were implemented using SPSS v.19. Results:385 cases diagnosed between 1993 and 2008 were included in the analysis. 55.8% of cases were male and 128 (33.2%) had EBV-associated disease. Median follow up was 74 months (range 0–220 months). Clinical stage was available for 289 patients: stage 1 (17.9%); stage 2 (41.8%); stage 3 (22.5%); and stage 4 (17.6%). In the analysis of all cases there were no significant differences in OS and EFS by number or presence of HLA-A*02:01 and A*01:01 alleles. EBV-associated cHL cases demonstrated inferior OS with the presence of HLA-A*02:01 (p=0.04) (figure). This was corroborated by the demonstration of inferior OS with increasing number of HLA -A*02:01 alleles present (p=0.016). Increasing number of HLA -A*02:01 alleles was also associated with inferior EFS but differences were not statistically significant. HLA-A*01:01 was not associated with OS or EFS in EBV-associated cHL. There was no significant association between histological subtype and OS or EFS. When clinical stage, age, and sex were included in the Cox-regression analysis of survival in EBV-associated cases, only age (p<0.001) was significant; presence of an HLA-A*02:01 allele approached statistical significance (p =0.062). Conclusions:In this study the HLA-A*02:01 allele was associated with inferior survival in EBV-associated cHL. As HLA-A*02:01 is associated with a decreased risk of developing EBV-associated cHL, we hypothesise that cHL arising in this 'protected' group is associated with greater biological or immunological dysfunction, resulting in poorer outcome. HLA-A genotype may be helpful in identifying patients who have inferior outcomes using standard therapy, and who may benefit from novel or cellular therapies. Larger studies within the context of clinical trials are required to extend these findings. In addition, this study supports the importance of stratifying cHL cases by EBV-status in clinical studies. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Epstein-Barr virus infection and expression of B-cell oncogenic markers in HIV-related diffuse large B-cell Lymphoma.

Epstein-Barr virus (EBV)-mediated lymphomagenesis in the setting of HIV infection has been widely accepted. However, little is known about how EBV impacts prognosis. We investigated the hypothesis that EBV infection is associated with expression of specific B-cell oncogenic markers in HIV-related diffuse large B-cell lymphoma (DLBCL) and examined the prognostic use of detecting EBV infection. HIV-related DLBCL cases diagnosed between 1996 and 2007 within Kaiser Permanente California were identified. Immunohistochemical staining was used to analyze the expression of selected markers that are cell-cycle regulators, B-cell activators, and antiapoptotic proteins among others. EBV infection was determined by in situ hybridization of EBV RNA. Correlations between EBV and marker expression were examined using Spearman correlation coefficient. The prognostic use of EBV status was examined in multivariable Cox model adjusting for International Prognostic Index (IPI). Receiver-operating characteristics (ROC) analysis was used to evaluate improvement in model discrimination. Seventy HIV-related DLBCL cases were included (31% EBV±). EBV+ tumor was associated with increased expression of BLIMP1 and CD30 and reduced expression of BCL6 and LMO2. EBV+ tumor was independently associated with elevated 2-year overall mortality [HR, 3.3; 95% confidence interval (CI), 1.6-6.6]. Area under the ROC curve showed improved model discrimination when incorporating tumor EBV status with IPI in the prediction model [0.65 vs. 0.74 (IPI only)]. Our results suggest that EBV infection was associated with expression of several tumor markers that are involved in the NF-κB pathway and that detecting tumor EBV status may have prognostic use in HIV-related DLBCLs.