EBV-positive Hodgkin Lymphoma Research Articles

Free Circulating Epstein Barr Virus DNA Load In Indian Children With Hodgkin Lymphoma As a Biomarker Of EBV-Associated Disease

IntroductionEpstein-Barr virus (EBV) association has been described in a large majority of children with Hodgkin lymphoma (HL) in India. The significance of circulating plasma EBV load and its kinetics during HL therapy is largely unknown. This study aimed at assessing the value of circulating EBV-DNA as a biomarker of EBV-associated pediatric HL and of tumour burden, and the value of serial monitoring during therapy. MethodAll pediatric cases presenting with lymphadenopathy were prospectively recruited between 2007 and 2012. Lymph node biopsy was performed. Children with HL and controls with reactive nodal hyperplasia were enrolled in the study after prior informed consent. Untreated non-lymphoid malignancies and healthy controls were also included in the control group. Plasma EBV real-time quantitative-PCR (RQ-PCR) was assessed with LightCycler2.0, Roche. EBV-associated HL was defined by positive EBV latent membrane protein-1 immunohistochemistry on lymph-node biopsies. Risk-adapted ABVD chemotherapy was given to HL cases, and early treatment response was assessed. RQ-PCR was repeated after the first cycle, at the end of treatment and on follow-up. Kaplan-Meier survival analysis was done, including loss to follow-up as an event. ResultsThirty cases of HL (newly diagnosed–28, relapses–2) and 70 age/ sex-matched controls (benign lymphadenopathy–19, non-lymphoid malignancy–29, Burkitt lymphoma–5, healthy children–17) were included. HL stage distribution was stage I–4, II–9, III–4 and IV–13. EBV immunohistochemistry was positive in 16 (59.3%) out of 27 HL cases analyzed (14/19 MC, 1/6 NS, 0/2 LP, 1 unclassified). RQ-PCR was detectable in 19 (63.3%) out of 30 HL, with 87.5% accuracy (Kappa coefficient=0.69 [0.42-0.97]). All 70 controls were RQ-PCR negative (p<0.0001). RQ-PCR sensitivity and specificity in EBV-associated HL detection were 87.5% and 81.8% respectively. Three out of 4 cases with more than 10,000 EBV copies/mL had advanced stage disease (III-IV) and B symptoms. The highest viral load (430,000 copies/mL) was seen in a boy with stage IV-B disease and end-stage liver involvement. However, viral load was not significantly associated with tumor burden or with survival. All treated Q-PCR positive cases showed EBV clearance after the first cycle. One case, RQ-PCR negative 10 years after primary treatment of stage-I HL, relapsed locally and became RQ-PCR positive. Five-year overall survival (OS) and event-free survival (EFS) were 82.6±8.5% and 77.2±8.3% respectively. OS and EFS were not significantly different in EBV-positive and EBV-negative HL. ConclusionRQ-PCR detection of circulating EBV-DNA is a biomarker of EBV-associated HL, in contrast with pediatric non-lymphoid malignancies, sporadic Burkitt lymphoma and benign lymphadenopathies. EBV-DNA may be used as an early marker of response to therapy in EBV-associated HL and may increase in case of relapse. Disclosures:No relevant conflicts of interest to declare.

Exposure to UV radiation and risk of Hodgkin lymphoma: a pooled analysis

AbstractUltraviolet radiation (UVR) exposure has been inversely associated with Hodgkin lymphoma (HL) risk, but only inconsistently, only in a few studies, and without attention to HL heterogeneity. We conducted a pooled analysis of HL risk focusing on type and timing of UVR exposure and on disease subtypes by age, histology, and tumor-cell Epstein-Barr virus (EBV) status. Four case-control studies contributed 1320 HL cases and 6381 controls. We estimated lifetime, adulthood, and childhood UVR exposure and history of sunburn and sunlamp use. We used 2-stage estimation with mixed-effects models and weighted pooled effect estimates by inverse marginal variances. We observed statistically significant inverse associations with HL risk for UVR exposures during childhood and adulthood, sunburn history, and sunlamp use, but we found no significant dose-response relationships. Risks were significant only for EBV-positive HL (pooled odds ratio, 0.56; 95% confidence interval, 0.35 to 0.91 for the highest overall UVR exposure category), with a significant linear trend for overall exposure (P = .03). Pooled relative risk estimates were not heterogeneous across studies. Increased UVR exposure may protect against HL, particularly EBV-positive HL. Plausible mechanisms involving UVR induction of regulatory T cells or the cellular DNA damage response suggest opportunities for new prevention targets.

Lymphoma and Epstein−Barr virus DNA in blood during interleukin‐2 therapy in antiretroviral‐naïve HIV‐1‐infected patients: a substudy of the ANRS 119 trial

Interleukin-2 (IL-2) therapy increased CD4 cell counts and delayed antiretroviral therapy (ART) initiation in HIV-infected patients in the Agence Nationale de Recherche sur le SIDA et les Hépatites Virales (ANRS) 119 trial. However, four cases of lymphoma were reported. Epstein-Barr virus (EBV) replication is associated with an increased risk of lymphoma in immunocompromised patients. We assessed whether IL-2 had an impact on EBV replication and the development of lymphoma. A total of 130 ART-naïve patients were randomized to receive IL-2 therapy (n = 66) or no treatment (n = 64). Clinical data for patients with lymphomas were reviewed and tumours assessed for evidence of EBV infection and CD25 (the IL-2 receptor) expression. EBV DNA levels were measured in whole blood and plasma in both arms using real-time polymerase chain reaction (PCR), up to 48 weeks after baseline (BL). Four lymphomas occurred, a median of 61 weeks [range 40-94 weeks] after randomization at a median CD4 cell count of 396 cells/μL (IQR 234-536 cells/μL). In the IL-2 arm, two patients developed EBV-positive Hodgkin's lymphoma, and one developed EBV-negative Burkitt-type lymphoma. One patient in the control group developed EBV-positive non-Hodgkin's lymphoma. CD25 was negative in all cases. Among the 41 of 55 (control arm) and 44 of 58 (IL-2 arm) patients with detectable EBV DNA in whole blood at both BL and week 48, the median change in EBV DNA between BL and week 48 was +0.04 log10 copies/ml in both arms (P = 0.7). In plasma, EBV was detected at least once in 22 of 52 controls and 21 of 54 IL-2-treated patients (P = 0.8). IL-2 therapy had no significant effect on EBV replication over 48 weeks in these ART-naïve patients. The occurrence of lymphomas did not seem to be associated with IL-2 therapy.

Suppression of the LMP2A target gene, EGR‐1, protects Hodgkin's lymphoma cells from entry to the EBV lytic cycle

Hodgkin's lymphoma is unusual among B cell lymphomas, in so far as the malignant Hodgkin/Reed-Sternberg (HRS) cells lack a functional B cell receptor (BCR), as well as many of the required downstream signalling components. In Epstein-Barr virus (EBV)-positive cases of Hodgkin's lymphoma, HRS cells express the viral latent membrane proteins (LMP)-1 and -2A. LMP2A is thought to contribute to the pathogenesis of Hodgkin's lymphoma by providing a surrogate BCR-like survival signal. However, LMP2A has also been shown to induce the virus-replicative cycle in B cells, an event presumably incompatible with lymphomagenesis. In an attempt to resolve this apparent paradox, we compared the transcriptional changes observed in primary HRS cells with those induced by LMP2A and by BCR activation in primary human germinal centre (GC) B cells, the presumed progenitors of HRS cells. We found a subset of genes that were up-regulated by both LMP2A expression and BCR activation but which were down-regulated in primary HRS cells. These genes included EGR1, an immediate-early gene that is required for BCR-induced entry to the virus-replicative cycle. We present data supporting a model for the pathogenesis of EBV-positive Hodgkin's lymphoma in which LMP2A-expressing HRS cells lacking BCR signalling functions cannot induce EGR1 and are consequently protected from entry to the virus lytic cycle. The primary microarray data are available from GEO (http://www.ncbi.nlm.nih.gov/geo/) under series Accession No 46143.

Changing trend of Epstein-Barr virus association in Hodgkin lymphoma in the Republic of Korea

Hodgkin lymphoma (HL) in resource-poor population shows an early childhood peak with predominance of mixed cellularity (MCHL) and high Epstein-Barr virus (EBV) association, whereas HL in resource-rich population has a peak in ages 15-29 years with the predominance of nodular sclerosis (NSHL) and low EBV association. We examined the changing trend of HL in association with EBV in Korea in 385 cases of HL (1980-2011). Data from 1980-1995 and 1996-2011 were compared. Age distribution showed a marked decrease in children (age < 15 years) (21.0 % to 8.6 %, p = 0.004) and sharp increase in young adults (ages 15-29 years) and ages 30-59 years. Male predominance decreased from 3.04 to 1.57. Predominant subtype changed from MCHL to NSHL; MCHL fell from 53.3 % to 26.4 % (p < 0.001), and NSHL rose from 24.8 % to 58.2 % (p < 0.001). EBV-positive HL decreased from 66.7 % to 38.2 %. However, in young adult females, EBV positive cases increased, contributing to the peak in this age group. Epidemiology of HL in Korea is rapidly changing into a Western pattern concurrent with the socioeconomic changes. Analysis of the cases stratified by EBV status and sex identified a smaller hidden peak in the young adult age, which confirms the four disease hypothesis proposed by Jarrett. We propose EBV testing as an essential component in epidemiologic study of Hodgkin lymphoma.

Abstract 4782: Detection of EBV nucleic acid and expression of oncogenic protein LMP-1 in Reed-Sternberg cells of Hodgkin's lymphoma.

Abstract Epstein-Barr virus (EBV) is the causative agent of infectious mononucleosis and is implicated in the development of several human malignancies, such as nasopharyngeal carcinoma (NPC) and Hodgkin's Lymphoma (HL). While the incidence of Hodgkin's Lymphoma in Iraq is not yet investigated, according to the WHO Hodgkin's Lymphoma is one of the 10 most frequent types of cancer in Iraq (http://www.who.int/disasters/repo/10062.pdf). The present study aimed to determine the association between HL and EBV in middle Euphrates region of Iraq, and to determine whether it is related to certain age group or specific histologic subtypes. It also aims to compare the pattern with other developing or developed countries. This retrospective study includes 86 formalin-fixed paraffin-embedded tissue blocks from HL patients, these were compared with 35 patients with Reactive Follicular Hyperplasia, that were age and gender matched. All the samples belonged to the period between Jan.2008 to April2010. The presence of EBV DNA was investigated by polymerase chain reaction (PCR), while the expression of the oncogenic protein (LMP-1) in Reed-Sternberg cells by immunohistochemistry (IHC). The mean age group of HL patients was 28.5 old years with a median of 25 years old. There were 49 males and 37 females. Age group of 16-45 years was the largest one and represented 66.2%. The third decade took the peak incidence (26/86, 30.2%). patients aged less than 16 years were 17.5%, while 16.3% of patients were more than 45 years old. Mixed cellularity HL was the most common subtype represented 47.6% of all the cases, followed by nodular sclerosis (33.7%) while the lymphocyte-rich, and lymphocyte depleted subtypes were (8.1%, 7%) respectively, whereas Lymphocyte Predominant was seen in 3.48% of cases. EBV DNA was detected in 39.5% of the cases, while the expression of LMP-1 in Reed Sternberg cells was found in 85.3% of the EBV-positive HL. The highest rate of EBV was seen in the children age group (less than 16 years; 53.3%), while the lowest rate was among young adults (16-45 years; 35.08%). Older age group (more than 45 years) has a rate of EBV presented (39.5%) which was higher than the young adult patients, but lower than the children age group. Histologically, the most frequent viral association was observed in mixed cellularity subtype (56.1%), followed by lymphocyte-rich and lymphocyte depleted subtypes (42.9%, 33.3%) respectively. The least frequency of association was in nodular sclerosis type (20.7%), while lymphocyte predominant subtype showed negative relationship with EBV-positive HL. EBV positive cases were mostly males (40.81%) compared to females (37.8%). Citation Format: Dhiya S. Al-Shumary, Karrar A.A. Ali Beg, Alae A. Yaseen, Salman A. Al-Jibouri, Kaswer M. Al-Toriahi. Detection of EBV nucleic acid and expression of oncogenic protein LMP-1 in Reed-Sternberg cells of Hodgkin's lymphoma. [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 4782. doi:10.1158/1538-7445.AM2013-4782 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

Atypical prediagnosis Epstein-Barr virus serology restricted to EBV-positive Hodgkin lymphoma

AbstractAn altered anti–Epstein-Barr virus (EBV) serologic profile preceding diagnosis is associated with an increased risk of Hodgkin lymphoma. It is unknown whether this atypical pattern predicts Hodgkin lymphoma risk further subdivided by determination of EBV in tumor cells. A nested case-control study of 128 incident Hodgkin lymphoma cases and 368 matched controls from active-duty military personnel with archived serum in the US Department of Defense Serum Repository was conducted to determine whether a panel of anti-EBV antibody titers differed in EBV+ and EBV− Hodgkin lymphoma. Among 40 EBV+ Hodgkin lymphoma cases and matched controls, statistically significant increased risks were associated with elevated anti-EBV VCA IgG antibody titers (relative risk = 3.1; 95% confidence interval [CI], 1.1-8.7), and an anti–EBNA-1/anti–EBNA-2 antibody ratio ≤ 1.0 versus > 1.0 (relative risk = 4.7; 95% CI, 1.6-13.8). In contrast, no significant associations were found among 88 EBV− Hodgkin lymphoma cases relative to their matched controls. In case-case analysis, EBV+ disease was significantly associated with a low anti–EBNA-1/anti–EBNA-2 antibody ratio. This distinc-tive serologic response to EBV latent antigens, indicative of immune dysfunction in other clinical settings, is associated with an increased risk of developing EBV+ but not EBV− Hodgkin lymphoma.

Impact of the Epstein–Barr virus positivity on Hodgkin's lymphoma in a large cohort from a single institute in Korea

Epstein-Barr virus (EBV) is considered a prognostic marker in Hodgkin lymphoma (HL) patients, but previous studies have yielded mixed findings because of the confounding effects of factors including age. We examined the prognostic impact of EBV status on 159 patients with HL. The median age at diagnosis was 32 years (range, 4-77 years). The median follow-up time was 5.83 years (range, 0.33-19.69 years). Tumor cell EBV status was positive in 34.5 %. EBV-positive HL was associated with age of ≥ 25 years, male gender, B symptoms, advanced stage, high-risk IPS, nonnodular sclerosis subtype, and treatment with chemotherapy only (P < 0.05). The 5-year disease-specific survival (DSS) rates were 94.1 and 76.4 % for the EBV-negative and EBV-positive HL, respectively, (P < 0.001). On univariate analysis, event-free survival, DSS, and overall survival (OS) were significantly associated with age 40 years or older, B symptoms, and high-risk international prognostic score (≥ 4). On multivariate analysis, EBV positivity was found to be a significant prognostic factor for DSS, particularly in adults 25 years or older. Subgroup analysis showed significant association of EBV-positive HL with poorer DSS and OS in adults 25 years or older with advanced stage disease. In the present series of HL patients, the presence of EBV in tumor cells is associated with adverse prognostic factors. EBV-positive HL is significantly associated with poorer DSS in all age groups.

EBNA3B-deficient EBV promotes B cell lymphomagenesis in humanized mice and is found in human tumors.

Epstein-Barr virus (EBV) persistently infects more than 90% of the human population and is etiologically linked to several B cell malignancies, including Burkitt lymphoma (BL), Hodgkin lymphoma (HL), and diffuse large B cell lymphoma (DLBCL). Despite its growth transforming properties, most immune-competent individuals control EBV infection throughout their lives. EBV encodes various oncogenes, and of the 6 latency-associated EBV-encoded nuclear antigens, only EBNA3B is completely dispensable for B cell transformation in vitro. Here, we report that infection with EBV lacking EBNA3B leads to aggressive, immune-evading monomorphic DLBCL-like tumors in NOD/SCID/γc-/- mice with reconstituted human immune system components. Infection with EBNA3B-knockout EBV (EBNA3BKO) induced expansion of EBV-specific T cells that failed to infiltrate the tumors. EBNA3BKO-infected B cells expanded more rapidly and secreted less T cell-chemoattractant CXCL10, reducing T cell recruitment in vitro and T cell-mediated killing in vivo. B cell lines from 2 EBV-positive human lymphomas encoding truncated EBNA3B exhibited gene expression profiles and phenotypic characteristics similar to those of tumor-derived lines from the humanized mice, including reduced CXCL10 secretion. Screening EBV-positive DLBCL, HL, and BL human samples identified additional EBNA3B mutations. Thus, EBNA3B is a virus-encoded tumor suppressor whose inactivation promotes immune evasion and virus-driven lymphomagenesis.

Sequential monitoring of plasma EBV-DNA level in a patient with EBV-positive Hodgkin lymphoma

Sequential monitoring of plasma EBV-DNA level in a patient with EBV-positive Hodgkin lymphoma

Radiotherapy-induced hyperthyroidism in a cystinotic kidney transplant patient with Hodgkin lymphoma

Hypothyroidism is a well-known complication of cystinosis. PTLD incidence in pediatric renal transplant population ranges between 1 and 4.5%. We describe the case of a young cystinotic patient who developed hyperthyroidism after radiotherapy for Hodgkin lymphoma. He is a 23-yr-old male who was diagnosed with cystinosis at the age of two. He developed renal failure and other extrarenal complications but never presented hypothyroidism. At the age of 12, he received a successful kidney transplant from a cadaveric donor. Two yr later, EBV-positive Hodgkin lymphoma was diagnosed and chemotherapy and radiotherapy were administered. He achieved remission. Eight yr later, autoimmune hyperthyroidism secondary to previous radiation was detected, and he slowly became symptomatic. Clinical symptoms and laboratory data spontaneously normalized. This is the first case of a cystinotic patient developing hyperthyroidism. Thyroid disorders, especially hypothyroidism, have been reported in association with neck irradiation. Hypothyroidism would have been considered to be a late complication of cystinosis and not a consequence of radiotherapy. Thyroid hormones, clinical examination, and history evaluation for thyroid dysfunction should be periodically monitored after neck radiotherapy. The thyroid should always be excluded from the irradiation fields. Multidisciplinary interaction in difficult cases should be encouraged.

Comparison of Clinical Characteristics, Plasma Cytokine Signature, Cytokine Gene Polymorphisms and Outcome Between EBV Positive and Negative Hodgkin Lymphoma: a Study of the Groupe Des Lymphomes De l'Adulte (GELA)

AbstractAbstract 3884 Background:Hodgkin Lymphoma (HL) has a remarkable pathogenesis with rare neoplastic cells surrounded by numerous reactive cells that interact with cytokines, chemokines and soluble receptors. EBV is found in about 40% of HL. Recent clinical series showed that prognostic between EBV-positive and negative could be different especially in patients older than 50 years old, with a worse prognosis for patients with EBV-positive tumors (Diepstra et al. JCO 2009). Gene expression profile analysis of HL tumors are also in favor of different pathogenesis between EBV-positive and negative tumors. In this context, we assessed whether plasma cytokine pattern and cytokine gene polymorphisms could be different regarding EBV status and compared the clinical characteristics and outcome of patients according to EBV status in a series of 242 HL patients. Patients and Methods:the GELA conducted between 1998 and 2002, a prospective study to assess the prognostic role at diagnosis of plasma levels of TNFA, its soluble receptors TNF-R1, TNF-R2, and IL-10, IL-6, IL-1B, IL-1Ra, sCD30 (Casasnovas et al. JCO 2007). This study was also designed to investigate the role of polymorphisms (SNP) in cytokine genes of IL10 (Four SNPs), TNFA, IL6, IL1B, IL1RN, INFG, CCL17 and IL12. EBV status of classical HL samples could be obtained retrospectively for 242 patients. Results:EBV status was obtained according to the immunohistochemistry detection of latent membrane protein-1 (LMP-1) in 186 patients (77%), to EBER in situ hybridation results in 48 patients (20%) or others techniques in the 8 remaining patients (3%). EBV status was considered as positive and negative for 53 (21%) and 189 (79%) patients, respectively. Comparison of clinical characteristics between EBV-negative and positive HL showed some differences in term of sex-ratio and histological subtypes with a higher proportion of male in EBV-positive cohort (81% vs. 54%, P < .01) and less nodular sclerosis subtypes (74% vs. 85% P = .05). Erythrocyte sedimentation rate and white-cell count were more frequently increased in EBV-negative population but the distribution of prognostic categories according to the Hasenclever score was not different between the two cohorts. The TNFA and TNF-R2 plasma levels were significantly higher for EBV-positive compared to EBV-negative HL (P = .03) but no difference was observed for others cytokines (TNF-R1, IL-10, IL-6, IL-1Ra, sCD30). For the eleven studied SNPs, no difference in term of genotype distribution was observed between EBV-positive and negative tumors. The 6-year overall survival (OS) (91% vs. 88% P = .3) and progression free survival (PFS) (81% vs. 79% P = .3) were not statistically different between the EBV-negative and positive cohorts. Analysis of outcome by age groups, showed a non-significant trend for a worse 6-year OS (72.1% vs. 58.3% P = .14) and 6-year PFS (57.2% vs. 48% P = .42) for EBV-positive HL after 50 years old with a similar proportion of EBV-positive and negative HL tumors before and after 50 years old. Conclusions:In this study, we confirm difference of histological subtype distribution between EBV-positive and negative HL. A difference of plasma levels of TNFA and TNF-R2 was observed suggesting a different inflammatory profile between EBV-positive and negative EBV HL but genotype distribution of germline cytokine gene polymorphisms was not different. In this cohort of HL with a global favorable outcome, a trend for a worse prognosis became apparent for older patients with EBV-positive HL. Disclosures:Salles:Roche: Consultancy, Honoraria.

Dominant expression of survival signals of endoplasmic reticulum stress response in Hodgkin lymphoma

The accumulation of viral proteins in endoplasmic reticulum (ER) may cause ER stress responses and lead to either apoptosis or survival depending on the driving signals. The strong expression of latent membrane protein-1 (LMP1) in Epstein-Barr virus (EBV)-positive Hodgkin lymphoma (HL) cells raises the question whether LMP1-induced ER stress response is associated with the characteristic tumor biology in HL. In this study, we investigated the expression of ER stress signals (glucose-regulated protein 78 [GRP78], X-box binding protein 1 [XBP1], activating transcription factor 6 [ATF6], CCAAT enhance-binding protein homologous protein [CHOP] and phospho-apoptosis signal-regulating kinase 1 [pASK1]) on 156 cases of HL. Furthermore, LMP1 transfection on EBV-negative HL cell lines was used to explore the regulation of ER stress signals by EBV-LMP1. Interestingly, we demonstrated that the survival signals of ER stress response (GRP78, 62%; XBP1u [unspliced], 55%; XBP1s [spliced], 38%; ATF6, 91%) were dominantly expressed over the ER death signals (CHOP, 10%; pASK1, 7%) in all histological subtypes of HL with a similar level in both EBV-positive and EBV-negative cases. However, expression of ER signals did not bear prognostic significance. In vitro, LMP1 transfection increased the expression of GRP78 and XBP1, but attenuated the expression of death signals, CHOP and pASK1. These data indicate that EBV-LMP1 may play a role in shifting EBV-infected cells towards the survival pathway in the presence of ER stress in EBV-positive HL cases.

Plasma EBV-DNA monitoring in Epstein-Barr virus-positive Hodgkin lymphoma patients

Epstein-Barr virus (EBV) is associated with approximately one-third of Hodgkin lymphoma (HL) cases. EBV-DNA is often present in the plasma and whole blood of EBV-associated HL patients. However, the significance of EBV-DNA monitoring is debated. In a cohort of 165 adult HL patients, EBV-DNA viral load was prospectively monitored both in the plasma and whole blood. Diagnostic tissue samples of all patients were histologically reviewed; in 72% nodular sclerosis was detected, 24% presented with mixed cellularity (MC), and 5% had other type of HL. Tissues from 150 patients were also analyzed for the presence of latent EBV infection using in situ hybridization for EBV-encoded RNA (EBER) and immunohistochemistry for latent membrane protein (LMP1). Using these methods, 29 (19%) patients were classified as EBV positive. Using real-time quantitative PCR, 22 (76%) of EBV-positive HL patients had detectable EBV-DNA in the plasma and 19 (66%) patients in whole blood prior to therapy. In the group of EBV-negative HL cases, three (2%) patients had detectable plasma EBV-DNA and 30 (25%) patients whole blood EBV-DNA before treatment. EBV-positive HL was significantly associated with EBV-DNA positivity both in the plasma and whole blood in pretreatment samples, increasing age and MC subtype. Serial analysis of plasma EBV-DNA showed that response to therapy was associated with decline in viral load. Moreover, significantly increased plasma EBV-DNA level recurred before disease relapse in one patient. Our results further suggest that the assessment of plasma EBV-DNA viral load might be of value for estimation of prognosis and follow-up of patients with EBV-positive HL.

Hodgkin lymphoma in a young child contributing to a diagnosis of ataxia telangiectasia: review of the literature

Ataxia telangiectasia (A-T) is a rare genetic disorder characterized by progressive cerebellar ataxia, oculocutaneous telangiectasias, immunodeficiency, chromosomal instability, and radiation sensitivity (Peterson et al. Lancet 283:1189–1193, 1964; Boder and Sedgwick Pediatrics 21:526–554, 1958; Taylor et al. Nature 258:427–429, 1975). Compared to the general population, patients with primary immunodeficiencies such as A-T have an increased rate of malignancy and an earlier age at presentation (Loeb et al. J Pediatr Hematol/Oncol 22:464–467, 2000; Taylor et al. Blood 87:423–438, 1996). We report the clinical, histopathologic, and molecular features of a 6-year-old child who presented with EBV-positive Hodgkin lymphoma (HL), which led to the diagnosis of ataxia telangiectasia. The diagnosis of HL at this unusually young age prompted further clinical, immunologic and cytogenetic evaluations, all of which supported a diagnosis of A-T. Because A-T patients are exquisitely sensitive to radiation, the patient was put on a modified chemotherapeutic regimen; however, 14 months later, he experienced a relapse and passed away by age 9. Classical HL is relatively rare among A-T patients, and this is the first documented case of an EBER-1 positive Hodgkin lymphoma in an ataxia telangiectasia patient. A review of the literature examining cases of HL in A-T is provided.

EBV-positive Hodgkin lymphoma is associated with suppression of p21cip1/waf1 and a worse prognosis

BackgroundAbout 30-50% of Hodgkin lymphomas (HLs) harbor the Epstein-Barr virus (EBV), but the impact of EBV infection on clinical outcomes has been unclear. EBV-encoded small RNAs (EBERs) are presented in all EBV-infected cells, but their functions are still less understood.ResultsEBER1 was transfected into two HL cell lines, KMH2 and L428, and microarrays were used to screen for EBER1-induced changes. We found that EBER1 suppressed p21cip1/waf1 transcription in HL cell lines. In addition, positive regulators of p21cip1/waf1 transcription, such as p53, EGR1, and STAT1, were decreased. Suppression of p21cip1/waf1 in the EBER1+ HL cell lines was associated with increased resistance to histone deacetylase inhibitors or proteasome inhibitors, drugs known to cause apoptosis by increasing p21cip1/waf1 levels. On biopsy specimens, EBV+ HLs had weaker expression of both p21cip1/waf1 and active caspase 3. Clinically, suppression of p21cip1/waf1 in EBV+ HLs was associated with a worse 2-year disease-free survival rate (45% for EBV+ HLs vs. 77% for EBV- HLs, p = 0.002).ConclusionAlthough the underlying mechanisms are still relatively unclear, EBER1 inhibits p21cip1/waf1 transcription and prevents apoptosis through down-regulation of p53, EGR1, and STAT1. The anti-apoptotic activity of EBER1 may be important in the rescue of Reed-Sternberg cells from drug-induced apoptosis and in the clinical behaviors of EBV+ HLs.

Risk of EBV-Positive Hodgkin Lymphoma Varies Over 30-Fold by HLA Class I Genotype and History of Infectious Mononucleosis.

Abstract 269 Introduction:Markers in the human leukocyte antigen (HLA) class I locus and infectious mononucleosis (IM) have both been associated with Epstein-Barr (EBV)-positive Hodgkin lymphoma (HL). IM is caused by late infection with EBV. To further explore associations between HLA class I genes and EBV-positive HL and to investigate potential interactions between genetic (HLA) and environmental risk factors in the development of HL, we performed a case-only study of 934 HL cases. Methods:HLA-A and B typing were performed by PCR sequence specific oligonucleotide assay. Logistic regression was used to estimate odds ratios (ORs) of EBV-positive HL relative to EBV-negative HL for HLA-A and B genotypes and other known risk factors. Results:Increasing age, male sex, HLA-A*01 and a previous history of IM were all associated with a statistically significant increased odds of EBV-positive HL while HLA-A*02 and HLA-B*07 were associated with a protective effect. Significant interactions were detected between HLA-A*01 and HLA-B*07 and between prior IM and HLA-A*02, such that HLA-B*07 abolished the increased odds associated with HLA-A*01 and HLA-A*02 abrogated the increased odds associated with prior IM. In our final model of EBV-positive HL, HLA-B*07 was associated with a dominant protective effect (OR=0.57, 95% confidence interval (CI): 0.34–0.94); HLA-A*02 was associated with a protective effect which was additive (for individuals with no prior IM, ORper allele =0.69, 95% CI: 0.50–0.97); and HLA-A*01 was associated with an increased odds in patients who were HLA-B*07-negative (ORper allele=2.15, 95% CI: 1.54–3.01). Prior IM was independently associated with EBV-positive HL (OR=3.35, 95% CI: 1.67–6.73). This resulted in a >32-fold higher odds of EBV-positive HL in HLA-A*01 homozygotes with IM compared to HLA-A*02 homozygotes without IM. Conclusions:We have identified strong associations between both genetic and environmental risk factors and EBV-positive HL. Our findings suggest that HLA class I-restricted, cell-mediated responses to EBV and a prior history of IM play critical roles in the pathogenesis of EBV-positive HL. Disclosures:No relevant conflicts of interest to declare.

Epstein-Barr-Virus-Infected CD15 (Lewis X)-Positive Hodgkin-Lymphoma-Like B Cells in Patients with Rheumatoid Arthritis

Patients with rheumatoid arthritis (RA), especially those who are treated with methotrexate (MTX), might have an increased risk of Hodgkin lymphoma (HL), a malignancy that is associated with Epstein-Barr virus (EBV). Here we describe a monoclonal EBV-infected B-lymphoblastoid cell line (LCL) called TKS-1 that was established from cells that spontaneously converted from an MTX-treated RA patient. TKS-1 has properties similar to HL cells and it is distinctly different from control LCLs established from normal individuals. TKS-1 cells express the HL -associated surface markers CD15 and CD30 (Takei et al. 1989). Like Hodgkin Reed-Sternberg (H-RS) cells of EBV-positive HL, TKS-1 cells express EBNA1 mRNA transcribed from the Qp promoter of the virus, whereas control LCLs use the Cp or Wp promoter to transcribe mRNA. TKS-1 cells can proliferate in an anchorage-independent manner and possess a cloning efficiency comparable to that of the Burkitt lymphoma (BL) line Raji. In addition, two EBV-positive LCLs established by cocultivated CD34+ cells isolated from the bone marrow of patients with RA and peripheral blood B lymphocytes from a healthy EBV-seronegative individual also expressed CD15. These results indicate that EBV-infected B-lymphoblastoid cells from patients with RA tend to acquire properties similar to HL cells.

Regulatory T cells and EBV associated malignancies

It has been shown that the T-regulatory cells (Tregs) not only play a key role in the establishment and maintenance of peripheral tolerance to prevent the autoimmune disease, but also inhibit the anti-tumor immunity. Recently, many studies have demonstrated that cytotoxicity T cells (CTL) can control the growth of EBV-positive tumor cells in vitro, including Hodgkin's lymphoma (HL), nasopharyngeal carcinoma, posttransplantation lymphoproliferative disorders (PTLD), depending on the large mount of EBV antigens presented by MHC molecules on the surface of these malignant cells. However, limited benefit of CTL adoptive immunotherapy has been reported in the treatment of EBV positive HL and NPC, and Tregs are regarded as a critical hurdle in this issue. In the present review, we discuss the correlation of EBV antigens expression in the tumor cells and the induction of Tregs in tumor microenvironment. Treg subsets and its possible mechanism to attenuate the anti-tumor immunity in EBV associated malignancies are also discussed, following by the possible strategies of targeting Tregs in the future immunotherapy for EBV positive cancers.

Racial/ethnic variation in EBV‐positive classical Hodgkin lymphoma in California populations

Epstein-Barr virus (EBV) is detected in the tumor cells of some but not all Hodgkin lymphoma (HL) patients, and evidence indicates that EBV-positive and -negative HL are distinct entities. Racial/ethnic variation in EBV-positive HL in international comparisons suggests etiologic roles for environmental and genetic factors, but these studies used clinical series and evaluated EBV presence by differing protocols. Therefore, we evaluated EBV presence in the tumors of a large (n = 1,032), racially and sociodemographically diverse series of California incident classical HL cases with uniform pathology re-review and EBV detection methods. Tumor EBV-positivity was associated with Hispanic and Asian/Pacific Islander (API) but not black race/ethnicity, irrespective of demographic and clinical factors. Complex race-specific associations were observed between EBV-positive HL and age, sex, histology, stage, neighborhood socioeconomic status (SES), and birth place. In Hispanics, EBV-positive HL was associated not only with young and older age, male sex, and mixed cellularity histology, but also with foreign birth and lower SES in females, suggesting immune function responses to correlates of early childhood experience and later environmental exposures, respectively, as well as of pregnancy. For APIs, a lack of association with birth place may reflect the higher SES of API than Hispanic immigrants. In blacks, EBV-positive HL was associated with later-stage disease, consistent with racial/ethnic variation in certain cytokine polymorphisms. The racial/ethnic variation in our findings suggests that EBV-positive HL results from an intricate interplay of early- and later-life environmental, hormonal, and genetic factors leading to depressed immune function and poorly controlled EBV infection.