RARE DISEASE
Epstein-Barr virus-positive diffuse large B-cell lymphoma
Epstein-Barr virus-positive diffuse large B-cell lymphoma
Epstein-Barr virus-positive diffuse large B-cell lymphoma
Synonyms: EBV-positive DLBCL, Epstein-Barr virus-positive diffuse large B-cell lymphoma not otherwise specified, Epstein-Barr virus-positive diffuse large B-cell lymphoma, NOS
Synonyms: EBV-positive DLBCL, Epstein-Barr virus-positive diffuse large B-cell lymphoma not otherwise specified, Epstein-Barr virus-positive diffuse large B-cell lymphoma, NOS
Synonyms: EBV-positive DLBCL, Epstein-Barr virus-positive diffuse large B-cell lymphoma not otherwise specified, Epstein-Barr virus-positive diffuse large B-cell lymphoma, NOS
Drug discovery
2
drugs
With orphan designations
Overview
Epstein-Barr virus-positive diffuse large B-cell lymphoma (EBV+ DLBCL) is an aggressive B-cell malignancy linked to EBV latency, accounting for 5–15% of DLBCL cases. Previously termed "EBV+ DLBCL of the elderly," it was reclassified in 2016 (WHO) due to occurrences in younger adults. Patients often present with extranodal involvement, B symptoms, and high-risk IPI scores. Prognosis is generally poor but varies regionally, with worse outcomes in Asian populations [1][4][10].
Population
Primarily affects older adults (>50 years), but cases reported in younger immunocompetent patients [1][6][15].
Higher prevalence in Asia and Latin America (up to 15% of DLBCL) compared to Western countries (4-7%) [4][7][14].
Associated with immunosenescence or immunosuppression (e.g., post-transplant) [2][10].
Therapies
First-line: R-CHOP (± etoposide) with variable response rates [1][8][9].
Novel approaches: PD-1 inhibitors (e.g., pembrolizumab), CD30-targeted therapies (brentuximab vedotin), and adoptive T-cell therapies under investigation [12][14].
Reduced-intensity regimens considered for frail patients [3][18].
Categories: rare hematological diseases, rare neoplastic diseases, rare transplant-related disorders
Research Papers
162 drug discovery papers about Epstein-Barr virus-positive diffuse large B-cell lymphoma, with 1 first-in-class and 2 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
162 drug discovery papers about Epstein-Barr virus-positive diffuse large B-cell lymphoma, with 1 first-in-class and 2 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
oligonucleotides
2026-06-28 | Epstein‐Barr Virus MicroRNAs as Key Regulators of Lymphoma Pathogenesis: Immune Evasion Mechanisms and Therapeutic Opportunities
The ubiquitous human gamma-herpesvirus Epstein-Barr virus (EBV) infects over 90% of adults globally and was the first human virus identified with oncogenic potential. EBV enters a lifelong persistence in the host via a finely regulated life-cycle comprising primary infection, latency and lytic reactivation. Within infected B-cells and epithelial cells, EBV encodes a distinct repertoire of microRNAs (miRNAs), primarily from the BART (BamHI A rightward transcript) and BHRF1 (BamHI H rightward open reading frame) clusters, which play pivotal roles in modulating both viral and host gene expression. These viral miRNAs contribute to key oncogenic processes: by dampening apoptotic responses (e.g., via targeting PUMA, Bim, and PTEN), promoting proliferation of latently-infected B-cells, inhibiting host immune responses (e.g., via down-regulation of CXCL-11 by miR-BHRF1-3), and promoting epithelial-mesenchymal transition (EMT) and metastasis through modulation of E-cadherin and other adhesion molecules. In human lymphomas, such as Burkitt lymphoma, Hodgkin lymphoma, and EBV-positive diffuse large B-cell lymphoma, the interplay of latent viral gene expression, miRNA-mediated regulatory networks, and host microenvironmental factors underlies malignant transformation and disease progression. Emerging evidence also supports the utility of EBV-encoded miRNAs as diagnostic and prognostic biomarkers in EBV-associated cancers. Importantly, therapeutic strategies aimed at interrupting viral miRNA function, restoring host tumor-suppressor pathways, and re-sensitizing tumor cells to immune surveillance hold promise. This review synthesizes current mechanistic insights into EBV-encoded miRNAs in oncogenesis, elaborates on their roles in lymphoma pathogenesis, and evaluates the translational potential of miRNA-targeted therapies in EBV-associated malignancies.
2023-09-08 | Alternative splicing of its 5′-UTR limits CD20 mRNA translation and enables resistance to CD20-directed immunotherapies
Aberrant skipping of coding exons in CD19 and CD22 compromises the response to immunotherapy in B-cell malignancies. Here, we showed that the MS4A1 gene encoding human CD20 also produces several messenger RNA (mRNA) isoforms with distinct 5' untranslated regions. Four variants (V1-4) were detected using RNA sequencing (RNA-seq) at distinct stages of normal B-cell differentiation and B-lymphoid malignancies, with V1 and V3 being the most abundant. During B-cell activation and Epstein-Barr virus infection, redirection of splicing from V1 to V3 coincided with increased CD20 positivity. Similarly, in diffuse large B-cell lymphoma, only V3, but not V1, correlated with CD20 protein levels, suggesting that V1 might be translation-deficient. Indeed, the longer V1 isoform contained upstream open reading frames and a stem-loop structure, which cooperatively inhibited polysome recruitment. By modulating CD20 isoforms with splice-switching morpholino oligomers, we enhanced CD20 expression and anti-CD20 antibody rituximab-mediated cytotoxicity in a panel of B-cell lines. Furthermore, reconstitution of CD20-knockout cells with V3 mRNA led to the recovery of CD20 positivity, whereas V1-reconstituted cells had undetectable levels of CD20 protein. Surprisingly, in vitro CD20-directed chimeric antigen receptor T cells were able to kill both V3- and V1-expressing cells, but the bispecific T-cell engager mosunetuzumab was only effective against V3-expressing cells. To determine whether CD20 splicing is involved in immunotherapy resistance, we performed RNA-seq on 4 postmosunetuzumab follicular lymphoma relapses and discovered that in 2 of them, the downregulation of CD20 was accompanied by a V3-to-V1 shift. Thus, splicing-mediated mechanisms of epitope loss extend to CD20-directed immunotherapies.
2022-03-24 | CircEAF2 counteracts Epstein-Barr virus-positive diffuse large B-cell lymphoma progression via miR-BART19-3p/APC/β-catenin axis.
Epstein-Barr virus (EBV) represents an important pathogenic factor of lymphoma and is significantly associated with poor clinical outcome of diffuse large B-cell lymphoma (DLBCL). Circular RNAs (circRNAs) play an essential role in lymphoma progression. However, the underlying mechanism of circRNA on DLBCL progression related to EBV remains largely unknown. CircRNA was screened by high-throughput sequencing in tumor samples of 12 patients with DLBCL according to EBV infection status. Expression of circEAF2, as well as the relationship with clinical characteristics and prognosis, were further analyzed in tumor samples of 100 DLBCL patients using quantitative real-time PCR. Gain- and loss-of-function experiments were conducted to investigate the biological functions of circEAF2 both in vitro and in vivo. The underlying mechanism of circRNA on DLBCL progression were further determined by RNA sequencing, RNA pull down assay, dual-luciferase reporter assay, rescue experiments and western blotting. We identified a novel circRNA circEAF2, which was downregulated in EBV + DLBCL and negatively correlated with EBV infection and DLBCL progression. In EBV-positive B lymphoma cells, circEAF2 overexpression induced lymphoma cell apoptosis and sensitized lymphoma cells to epirubicin. As mechanism of action, circEAF2 specifically targeted EBV-encoded miR-BART19-3p, upregulated APC, and suppressed downstream β-catenin expression, resulting in inactivation of Wnt signaling pathway and inhibition of EBV + DLBCL cell proliferation. In EBV-positive B-lymphoma murine models, xenografted tumors with circEAF2 overexpression presented decreased Ki-67 positivity, increased cell apoptosis and retarded tumor growth. CircEAF2 counteracted EBV + DLBCL progression via miR-BART19-3p/APC/β-catenin axis, referring circEAF2 as a potential prognostic biomarker. Therapeutic targeting EBV-encoded miRNA may be a promising strategy in treating EBV-associated lymphoid malignancies.
2021-07-08 | Deletion of Viral microRNAs in the Oncogenesis of Epstein–Barr Virus-Associated Lymphoma
Epstein–Barr virus (EBV), which encodes >80 genes and nearly 50 non-coding RNAs, is a double-stranded DNA virus. EBV is associated with various types of lymphomas and lymphoproliferative disorders not only of B-cell but also T/NK-cell origin. However, the oncogenic mechanism remains poorly understood, including the EBV receptors expressed on T/NK cells, relationship of EBV with host genes, and epigenetic regulation of EBV and host genes. The roles of host and viral non-coding RNAs during tumorigenesis have been elucidated. EBV encodes at least 49 mature microRNAs (miRNAs), of which 44 are located in Bam HI-A rightward transcripts (BARTs) region, and the remaining five are located in Bam HI-H rightward fragment 1. BART miRNAs modulate cell differentiation, proliferation, apoptosis, and the cell cycle, and they are considered positive regulators of oncogenesis. We and others have recently reported that EBV-positive lymphomas frequently possess large deletions in BART miRNA clusters, suggesting that some viral miRNAs have suppressive effects on oncogenesis, and that deletion of these miRNAs may aid lymphoma formation.
2020-03-24 | EBV microRNA-BHRF1-2-5p targets the 3'UTR of immune checkpoint ligands PD-L1 and PD-L2.
Epstein-Barr virus-positive (EBV+) diffuse large B-cell lymphomas (DLBCLs) express high levels of programmed death ligand 1 (PD-L1) and PD-L2. MicroRNA (miR) regulation is an important mechanism for the fine-tuning of gene expression via 3'-untranslated region (3'UTR) targeting, and we have previously demonstrated strong EBV miR expression in EBV+ DLBCL. Whereas the EBV latent membrane protein-1 (LMP1) is known to induce PD-L1/L2, a potential counterregulatory role of EBV miR in the fine-tuning of PD-L1/L2 expression remains to be established. To examine this, a novel in vitro model of EBV+ DLBCL was developed, using the viral strain EBV WIL, which unlike common laboratory strains retains intact noncoding regions where several EBV miRs reside. This enabled interrogation of the relationship among EBV latency genes, cell of origin (COO), PD-L1, PD-L2, and EBV miRs. The model successfully recapitulated the full spectrum of B-cell differentiation, with 4 discrete COO phases: early and late germinal center B cells (GCBs) and early and late activated B cells (ABCs). Interestingly, PD-L1/L2 levels increased markedly during transition from late GCB to early ABC phase, after LMP1 upregulation. EBV miR-BamHI fragment H rightward open reading frame 1 (BHRF1)-2-5p clustered apart from other EBV miRs, rising during late GCB phase. Bioinformatic prediction, together with functional validation, confirmed EBV miR-BHRF1-2-5p bound to PD-L1 and PD-L2 3'UTRs to reduce PD-L1/L2 surface protein expression. Results indicate a novel mechanism by which EBV miR-BHRF1-2-5p plays a context-dependent counterregulatory role to fine-tune the expression of the LMP1-driven amplification of these inhibitory checkpoint ligands. Further identification of immune checkpoint-targeting miRs may enable potential novel RNA-based therapies to emerge.
antibodies
2026-07-12 | Spatial Architecture of macrophages and T-cells Orchestrates Anti-Lymphoma Immunity and the Response to Immunochemotherapy in EBV+DLBCL
Epstein-Barr Virus (EBV)-positive Diffuse Large B-Cell Lymphoma (EBV+ DLBCL) is a rare and aggressive B-cell lymphoma characterized by heterogeneous pathological features and poor clinical outcomes. The cellular characteristics and spatial architecture of EBV+ DLBCL tumor microenvironment (TME) and its underlying mechanisms remain elusive. Twenty-one EBV+ DLBCL and 24 EBV-negative DLBCL (EBV- DLBCL) lymph node samples were analyzed using imaging mass cytometry (IMC) and the digital spatial profiler (DSP). Our findings revealed that EBV+ DLBCL exhibited a more suppressive TME with increased recruitment of CD163+ and IDO+ tumor-associated macrophages (TAMs) and more exhausted T cells compared to EBV- DLBCL. Consistently, spatial interaction analysis showed that T cells around TAMs displayed more exhausted phenotype. Additionally, more TAMs preemptively hijacked T cells before T-cell mediated attack in EBV+ DLBCL. Mechanistically, more TAMs recruited by EBV+ DLBCL resulted in upregulation of CCL5 through activation of JAK3/STAT3 pathway. Clinically, responders to immunochemotherapy exhibit higher levels of TAMs residing around tumor cells under T-cell attack compared to non-responders. Patients receiving immunomodulatory drugs (PD-1 inhibitors and lenalidomide) combined with immunochemotherapy seem to achieve better outcome. These insights enhance our understanding of the immune architecture in EBV+ DLBCL and identify new targets for immunotherapy, potentially improving patient outcomes.
2026-04-03 | Abstract 7218: Preliminary exploration of the synergistic mechanism between EBV LMP1 and MYD88 L265P mutation on the efficacy of PD-1 Inhibitors in ABC-DLBCL
Abstract Background: Activated B-cell-like diffuse large B-cell lymphoma (ABC-DLBCL) is aggressive with poor prognosis. Epstein-Barr virus (EBV) infection and the MYD88 L265P mutation are key molecular drivers in ABC-DLBCL, typically exhibiting mutual exclusivity, potentially due to functional overlap. PD-1 inhibitors show limited efficacy in unselected relapsed/refractory DLBCL patients. However, our retrospective analysis of relapsed/refractory non-GCB DLBCL patients suggested that rare "double-positive" (EBV-positive/MYD88 L265P mutant) patients had higher response rates to PD-1 inhibitors. Objective: This study investigated how EBV LMP1 modulates intracellular signaling in the context of MYD88 L265P mutation (causing constitutive JAK/STAT activation) and evaluated its synergistic effect with the PD-1 inhibitor tislelizumab. Methods: Clinical data from 54 relapsed/refractory non-GCB DLBCL patients treated with PD-1 antibodies were analyzed. Patients were stratified by EBER ISH (EBV) and MYD88 sequencing. The HBL-1 cell line (MYD88 L265P mutant, EBV-negative) was used. LMP1 was overexpressed via transfection. Groups: control, tislelizumab alone, LMP1 overexpression, LMP1+tislelizumab. qPCR measured mRNA levels of NF-κB p65, MMP9, c-Myc, TLR3, STAT3, JAK3, LMP1, PD-1, PD-L1. Western blot detected protein expression and phosphorylation of key molecules. Results: Among 21 patients with complete molecular data, both "double-positive" patients achieved an objective response (100%), compared to 22.2% in EBV-negative/MYD88 mutant (n=9) and 60.0% in EBV-positive/MYD88 wild-type (n=5) groups. HBL-1 cells showed high baseline p-JAK3 and p-STAT3. LMP1 overexpression suppressed p-JAK3 and p-STAT3. HBL-1 cells expressed tumor cell-intrinsic PD-1 mRNA. Tislelizumab alone inhibited p-JAK3/p-STAT3 phosphorylation. The combination of LMP1 overexpression and tislelizumab showed the strongest inhibition, indicating synergy. Conclusion: MYD88 L265P mutation may cause tumor cell addiction to STAT3 activation maintained by tumor cell-intrinsic PD-1 signaling. LMP1 co-existence adds inhibitory pressure, potentially enhancing this dependency. Tislelizumab blockade may yield a synthetic lethal effect. Combined EBV status and MYD88 profiling could predict PD-1 inhibitor sensitivity in DLBCL, requiring further validation. Citation Format: Chang Wang, Yingtao Lin, Jiesong Wang, . Preliminary exploration of the synergistic mechanism between EBV LMP1 and MYD88 L265P mutation on the efficacy of PD-1 Inhibitors in ABC-DLBCL [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7218.
2026-04-01 | Correction
Article title: Characteristics and prognosis of Epstein Barr virus-positive diffuse large B-cell lymphoma: a retrospective, single-center studyAuthors: Qiaochu, L., Lianming, L., Jianzhen, S., Xiao...
2026-03-07 | Epstein–Barr virus-associated lymphoma: current understanding and treatment strategies
Epstein-Barr virus (EBV) infection is well-known for its high prevalence rate, association with several diseases including cancer and autoimmune conditions, and a wide variety of symptoms and prognosis. When acquired at a young age, primary infections are often asymptomatic; however, in adolescence and young adulthood, symptomatic infections develop, such as in infectious mononucleosis. A special feature of EBV infection is its ability to establish a latent infection in B cells which can lead to long-term infection. Subsequent cellular transformation and viral protein expression can result to EBV-mediated carcinogenesis. Latent proteins expressed by EBV play a role in the pathogenesis of EBV infection and carcinogenesis. These proteins are responsible for a diverse range of functions including cell transformation, cell reprogramming, immune evasion, immune suppression, angiogenesis, cell cycle regulation, and B-cell receptor mimicry.EBV infection is associated with diffuse large B-cell lymphoma, Hodgkin lymphoma, NK/T cell lymphoma, Burkitt lymphoma, post-transplant lymphoproliferative disorders, and primary CNS lymphoma. The clinical presentation varies depending on the specific disease and EBV status, with EBV-positive lymphomas generally associated with poorer prognosis than EBV-negative cases.This review aimed to examine the current understanding of the pathogenesis of EBV-associated lymphoma and to evaluate emerging and accepted therapeutic strategies.
2026-01-09 | Rapidly progressive EBV-positive DLBCL arising from a duodenal diverticulum
Rapidly progressive EBV-positive DLBCL arising from a duodenal diverticulum An 81-year-old man with a history of diabetes mellitus presented with hematemesis.Urgent esophagogastroduodenoscopy (EGD) revealed an adherent blood clot inside a periampullary duodenal diverticulum, suggesting diverticular bleeding ( Fig. 1 A).Since no active bleeding was endoscopically observed, conservative therapy was adopted.Five days later, follow-up endoscopy showed an apparent benign ulcer inside the diverticulum ( Fig. 1 B).Two months later, the patient recurred melena and repeat EGD demonstrated a large ulcerative lesion surrounded by irregular elevations at the same site ( Fig. 1 C).Bioptic examination revealed diffuse infiltration of large atypical lymphoid cells in the stroma.Immunohistochemistry showed CK AE1/AE3(-), CD3(-), CD20( + ), CD5(-), CD10(-), BCL2( + ), BCL6(-), MUM1( + ), and EBER( + ) ( Fig. 2 ), establi-
small molecules
2026-07-28 | Clinical-Stage Nanatinostat Triggers Productive Epstein-Barr Virus Lytic Replication.
A phase 1b/2 study of the all-oral "kick-and-kill" combo nanatinostat + valganciclovir (nana-val) showed a ~40% overall response rate in patients with relapsed/refractory Epstein-Barr virus (EBV)-associated lymphomas. Despite the promising results in these difficult-to-treat cancers, there is little known about the effects of nanatinostat on EBV and cell death. In this work we demonstrate that nanatinostat, an HDAC1/3 inhibitor, robustly induces the full EBV lytic cycle in a panel of EBV-positive lymphoma and transformed cell lines (Burkitt, DLBCL, lymphoblastoid), spanning viral latency types and EBV genotypes. Nanatinostat drives lytic-gene transcription across all kinetic classes, viral genome replication, and virion release, and kills cells at nanomolar concentrations. Although valganciclovir does not increase cytotoxicity, its ability to block viral DNA synthesis and virion production justifies its inclusion to prevent viral dissemination. Importantly, the two main metabolites of nanatinostat lack histone-H3 acetylation activity and neither activate nor inhibit the EBV lytic phase, indicating that they are biologically inert with respect to EBV. These mechanistic insights clarify the clinical efficacy of nana-val and support further development of kick-and-kill strategies for EBV-driven malignancies.
2026-05-05 | Adult T-Cell Leukemia/Lymphoma and Epstein-Barr Virus-Positive DLBCL: A Rare Concomitant Association.
Adult T-cell leukemia/lymphoma (ATLL) is an aggressive lymphoma with a poor prognosis. The human T-lymphotropic virus 1 (HTLV-1) is associated with immunodeficiency and increased extranodal involvement in patients with diffuse large B-cell lymphoma (DLBCL). We report on a 47-year-old woman with spastic paraparesis and hepatitis B who was diagnosed with the acute form of ATLL. The clinical picture reveals peripheral generalized lymphadenopathy and splenomegaly. Findings on a hematologic exam indicated leukocytosis with lymphocytosis. A bone marrow biopsy/aspiration confirmed 50% T-cell lymphoid infiltration. Biochemistry results revealed hypercalcemia and a high lactate dehydrogenase value. Results of a CT scan indicated abdominal and thoracic adenopathy as well as moderate splenomegaly. A supraclavicular lymph node biopsy established a DLBCL diagnosis. The final diagnosis was composite lymphoma, DLBCL, and ATLL. The CHOP (cyclophosphamide, doxorubicin hydrochloride, vincristine sulfate [Oncovin], and prednisone) regimen was chosen due to the patient's ECOG performance status. Multiple infectious complications were diagnosed during chemotherapy-induced secondary aplasia. A complete remission, confirmed via PET-CT imaging, was obtained. After 1 month, a skin tumor on the upper right thigh was discovered and biopsied, and the histopathological exam and immunochemistry findings indicated Epstein-Barr virus-DLBCL lymphoma. The association of 2 aggressive lymphomas in a single HTLV-1 carrier is a rare report, and the evolution was severe, complicated by opportunistic infections, and unfavorable.
2026-01-16 | ORY1001 enhances anti-tumor immunity via CXCL10-mediated macrophage modulation in EBV-positive DLBCL.
Epstein-Barr virus-positive diffuse large B-cell lymphoma (EBV+ DLBCL) is an aggressive malignancy characterized by an immunosuppressive tumor microenvironment (TME), where tumor-associated macrophages (TAMs) often exhibit an M2-like phenotype that facilitates immune evasion. Here, we show that the LSD1 inhibitor ORY1001 reprograms the TME of EBV+ DLBCL through CXCL10-mediated modulation of macrophage polarization. Using a reductionist model in which EBV-negative DLBCL cell lines were transfected with LMP1, we demonstrate that ORY1001 induces robust expression of CXCL10, TNF-α and IL-6 in LMP1+ Riva cells, promoting M1 macrophage differentiation and reprogramming M2 macrophages toward a pro-inflammatory phenotype. This CXCL10-driven macrophage reprogramming was also observed in an additional LMP1+ DLBCL line (U2932) and in primary EBV+ DLBCL cells, in which ORY1001 selectively increased CXCL10 levels in tumor-cell supernatants, and conditioned media from these cultures—but not direct ORY1001 exposure—drove M1 polarization of THP-1 and primary CD14+ monocytes. These effects were dependent on CXCL10-CXCR3 signaling, as pharmacological inhibition of CXCR3 abrogated M1 polarization. In vivo, ORY1001 suppressed tumor growth in LMP1-expressing A20 lymphoma models, increased CXCL10 levels, and enhanced M1 macrophage infiltration. Furthermore, ORY1001 synergized with anti–PD-1 therapy to enhance T-cell activation, granzyme B expression, and tumor regression. These findings highlight LSD1 inhibition as a promising immunoepigenetic strategy to remodel the TME and improve the efficacy of immune checkpoint blockade in EBV-associated lymphomas.
2025-10-17 | Withaferin A inhibits EBV-driven lymphomagenesis through multiple mechanisms, including EBNA1 degradation.
Epstein-Barr virus (EBV) infects over 90% of the global population and drives multiple aggressive B-cell malignancies, including Burkitt lymphoma, diffuse large B-cell lymphoma, and Hodgkin lymphoma. Standard chemoimmunotherapy regimens can be highly effective, yet Epstein-Barr virus positive (EBV+) lymphomas sometimes exhibit poorer responses, higher resistance, and worse survival compared with Epstein-Barr virus-negative (EBV-) counterparts. This reflects the virus's ability to drive immune evasion, alter cell death pathways, and exploit host immune dysfunction, underscoring the potential value of EBV-directed strategies. Withaferin A (WA), a steroidal lactone with known anticancer and anti-inflammatory properties, was evaluated for its efficacy against EBV-associated B-cell non-Hodgkin lymphomas (B-NHL). Across a panel of lymphoma cell lines, WA demonstrated selective cytotoxicity toward EBV+ B-NHL, in part through proteasome-dependent degradation of EBNA1 (EBV nuclear antigen 1) and subsequent loss of viral episomes, alongside additional effects on cellular stress and survival pathways. Mechanistic studies revealed that WA collapses antioxidant defenses, drives oxidative stress, and suppresses NF-κB signaling, creating a multipronged disruption of viral and host survival pathways. In primary B-cell models and a cord blood-humanized mouse model of EBV-driven lymphomagenesis, WA inhibited B-cell transformation, reduced splenomegaly and tumor burden, and significantly prolonged survival without evidence of increased viral replication. These findings establish WA as a potent preclinical candidate that selectively targets vulnerabilities unique to EBV-transformed B cells, supporting further optimization and evaluation for EBV+ B-cell malignancies.
2025-09-30 | Immune Checkpoint Molecule Indoleamine 2,3-Dioxygenase 1 (IDO1) Is Expressed in Lymphoma Subtypes With and Without Epstein-Barr Virus.
Indoleamine 2,3-dioxygenase 1 (IDO1) is an immune regulator involved in innate and acquired immunity and immune escape of tumors. Its expression in the tumor microenvironment of several solid tumors and lymphomas associated with Epstein-Barr virus (EBV), together with the promising results from clinical trials of IDO1 inhibitors, prompted us to evaluate IDO1 expression in a large cohort of 455 lymphomas including 154 cases associated with EBV. We optimized an immunohistochemical assay to evaluate IDO1 staining and show that IDO1 expression in seen in several lymphoma subtypes including classic Hodgkin lymphoma (CHL, 40%), diffuse large B-cell lymphoma (DLBCL, 23.5%), lymphoproliferative disorders in post-transplant settings (LPD-PT, 71.4%), extranodal NK/T-cell lymphoma (ENKTL, 92%), and ALK-negative anaplastic large cell lymphoma (ALCL, 39%), among others. Multiplex immunofluorescence further aided in refining the localization of IDO1 protein expression particularly within the tumor microenvironment. There was a significant correlation between IDO1 expression and EBV positivity in CHL (83.8%), LPD-PT (86.2%), and ENKTL (91.5%), with statistically significant difference in the mean IDO1 H-scores between EBV-positive and EBV-negative cases. IDO1 expression in ALCL was confined to ALK-negative cases with a significant correlation between IDO1 expression and ALK status. Our findings show that IDO1 expression is not only highly correlated with lymphomas associated with EBV, but also found in lymphomas unassociated with EBV, including aggressive and refractory subtypes of lymphomas for which immune checkpoint inhibition through IDO1 could be exploited for therapeutic purposes.
cell therapies
2025-12-01 | Successful Use of Haploidentical HSCT in a Child With Schimke Immuno-Osseous Dysplasia Who Developed PTLD After Kidney Transplantation.
Schimke Immuno-Osseous Dysplasia Is a Rare Autosomal Recessive Multisystem Disorder Caused by Biallelic Pathogenic Variants in the SMARCAL1 Gene, Which Encodes a DNA Annealing Helicase Essential for Replication Fork Stability and Genomic Maintenance. Loss of SMARCAL1 Function Leads to Genomic Instability, Resulting in a Characteristic Clinical Triad of Disproportionate Short Stature, Steroid-Resistant Nephrotic Syndrome and Immunodeficiency. Kidney Transplantation Is the Standard Treatment for End-Stage Renal Disease in Schimke Immuno-Osseous Dysplasia. However, the Underlying Genomic Fragility and Immunodeficiency Heighten the Risk of Post-Transplant Complications, Particularly Epstein-Barr Virus-Associated Post-Transplant Lymphoproliferative Disorder. We Report the Case of a 7-Year-Old Girl With Genetically Confirmed Schimke Immuno-Osseous Dysplasia Who Developed Epstein-Barr Virus-Positive Diffuse Large B-Cell Lymphoma Eight Months Following Maternal Kidney Transplantation, Despite Reduced-Intensity Immunosuppression. Rituximab Therapy Resulted in Complete Remission, but Required the Withdrawal of all Immunosuppressive Agents, Thereby Posing a Risk of Kidney Graft Loss. In the Context of Primary Immunodeficiency and the Development of Post-Transplant Lymphoproliferative Disorder in Such a Setting, the Patient Subsequently Underwent Haploidentical Hematopoietic Stem Cell Transplantation From the Mother, Using an αβ T-Cell-Depleted Graft and a Nephrotoxicity-Sparing Conditioning Regimen. Full Donor Chimerism Was Achieved by Day 28. Immune Reconstitution Occurred Within One Year. At 27 Months After Hematopoietic Stem Cell Transplantation She Remains in Remission With Preserved Kidney Function and no Ongoing Immunosuppression. This Case Highlights the Feasibility of Haploidentical Hematopoietic Stem Cell Transplantation as a Curative Salvage Strategy for Immunosuppression-Free Survival After Kidney Transplantation in Patients With Schimke Immuno-Osseous Dysplasia.
2025-08-01 | 28P Pre-clinical development of CAR-T cells targeting a lytic EBV antigen gp350 for therapy of EBV-positive diffuse large B-cell lymphoma
Epstein-Barr virus (EBV) associated diffuse large B-cell lymphoma (DLBCL) represents a rare aggressive B-cell lymphoma subtype characterized by an adverse clinical outcome. Despite intensive chemotherapy, the prognosis of EBV-positive DLBCL patients remains unfavorable, and a definitive method to completely eliminate latent EBV infection is lacking. Developing chimeric antigen receptor (CAR)-T cells therapy targeting EBV specific protein in the context of this EBV+ aggressive lymphoma will provide a novel therapeutic strategy for patients with limited treatment options.
2025-06-01 | 686 | TABELECLEUCEL IN EBV POSITIVE POST‐TRANSPLANT LYMPHOPROLIFERATIVE DISEASE: A REAL WORD EXPERIENCE
Introduction: ALLELE trial reported promising data on tabelecleucel, an on-demand allogeneic cytotoxic T-cell therapy for relapse/resistant epstein-barr positive post-transplant lymphoproliferative disease (EBV+ PTLD) that occurred after either solid organ (SOT) or hematopoietic transplantation (HCT), with a median OS improved from less than 6 to more than 18 months. The aim of this study is to evaluate feasibility and clinical outcome of PTLD in our center since the availability of tabelecleucel as compassionate use. Methods: Patients who developed histologically documented PTLD in adulthood after either SOT or HCT were analyzed since November 2023. Results: Between November 2023 and February 2025 we diagnosed 12 consecutive cases of PTLD, 10 after SOT (5 heart, 2 lungs, 1 kidney, 1 liver and 1 multivisceral transplant) and 2 after HCT. Median age at presentation of PTLD was 53 years (range 25–76), 8 patients were male. At diagnosis main adverse clinical features were: stage III–IV (n = 10, 83%), B symptoms (n = 3; 25%); ECOG > 2 (n = 7, 58%), bulky disease (2 patients, 16%). According to IPI score 50% of patients were identified as intermediate-high/high risk. Median time from transplant to PTLD was 86 months (range 3 to 372). We documented 9 DLBCL, 1 PEL, 1 polymorphic lymphoproliferative disorder and 1 primitive CNS lymphoma. EBV-DNA integration in lymphoma cells (EBER +) was documented in 9 patients (75%). All patients received Rituximab monotherapy: 2 patients achieve CR (16%) and did not require other additional therapy. Tabelecleucel was required as compassionate use in Rituximab Resistant EBV+ PTLD in 7 patients (2 after HCT and 5 not eligible/refusing chemotherapy program after SOT). In a median time of 14 days we were able to define the origin of the disease in 6 out of 7 cases (5 from patient, 1 from donor). In less than a week we identified a suitable batch of tabelecleucel in all the cases. One patient died before tabelecleucel delivery due to disease progression. Two patients are planned to be reinfused. For the remaining, median time from request to first Tabelecleucel dose was 22 days (range 11–44). Three patients completed tabelecleucel therapy, one is still under treatment. No hematological or extra hematological toxicities were documented. We documented one CR, one SD and one PD. This latter patient received R-CHOP achieving CR. After a median follow up of 8 months all the three patients are alive. The 3 patients with EBV negative Rituximab refractory PTLDs were treated with R-CHOP obtaining CR and are alive after a median follow up of 7 months. Conclusion: Our single center experience showed that tabelecleucel therapy is feasible in a real word setting. Time from “brain to vein” is short, thus preventing disease progression in the majority of cases. In our experience tabelectelucel can represent a good alternative to chemotherapy in EBV positive Rituximab resistant/refractory lymphoproliferative disorders. Keywords: cellular therapies Potential sources of conflict of interest: A. M. Barbui Consultant or advisory role: Pierre Fabre Honoraria: Pierre Fabre Educational grants: Pierre Fabre
2024-08-21 | Transition From Epstein-Barr Virus (EBV)-Positive Rectal Hodgkin Lymphoma to Diffuse Large B-Cell Lymphoma in the Lung.
We report a distinctive case of sequential lymphomas in a 72-year-old male, initially diagnosed with Epstein-Barr virus (EBV)-positive rectal classic Hodgkin lymphoma (cHL), followed by the development of diffuse large B cell lymphoma (DLBCL) in the lung. This rare progression underscores the complexity of lymphomas associated with EBV infection and their unpredictable clinical courses. The patient's journey began with symptoms of intractable diarrhea, low appetite, and significant weight loss, leading to the diagnosis of stage 4B cHL, managed initially with brentuximab/doxorubicin, vinblastine, dacarbazine (AVD) chemotherapy. Despite a partial response, surveillance identified a transition to DLBCL, marked by new pulmonary lesions. This case highlights the clinical and diagnostic challenges in managing sequential lymphomas, emphasizing the role of EBV in lymphomagenesis and the potential for clonal evolution from a common precursor cell. The therapeutic approach evolved from targeted chemotherapy to consideration of advanced treatments such as autologous stem cell transplant and chimeric antigen receptor (CAR) T-cell therapy, reflecting the aggressive nature and poor prognosis of the disease. This case contributes to our understanding of the EBV's impact on lymphoma progression and underscores the need for vigilant monitoring and adaptive treatment strategies in similar clinical scenarios.
2024-07-19 | Infant with diffuse large B-cell lymphoma identified postmortem with homozygous founder Slavic RAG1 variant: a case report and literature review.
There is an increased risk of lymphomas in inborn errors of immunity (IEI); however, germline genetic testing is rarely used in oncological patients, even in those with early onset of cancer. Our study focuses on a child with a recombination-activating gene 1 (RAG1) deficiency who was identified through a screening program for Slavic founder genetic variants among patients who died with malignancy at an early age in Belarus. We identified one homozygous founder RAG1 variant out of 24 available DNA samples from 71 patients who developed lymphoma aged <3 years from the Belarusian cancer registry between 1986 and 2023. Our patient had an episode of pneumonia at 3 months of age and was hospitalized for respiratory distress, candida-positive lung disease, and lymphadenopathy at 14 months of age. The diagnosis of Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) was established. The patient had a normal lymphocyte count that decreased over time. One month after chemotherapy initiation, the patient died due to sepsis and multiple organ failure without a genetic diagnosis. In a retrospective analysis, T-cell receptor excision circles (TRECs) and kappa-deleting recombination excision circles (KRECs) were undetectable in peripheral blood. A targeted screening program designed to detect a Slavic founder variant in the RAG1 gene among children revealed a 14-month-old Belarusian male infant with low TREC levels who died of EBV-driven DLBCL and complications of chemotherapy including infections. This case highlights how patients with IEI and recurrent infections may develop serious non-infectious complications, such as fatal malignancy. It also emphasizes the importance of early identification, such as newborn screening for severe combined immune deficiency. Earlier diagnosis of RAG deficiency could have prompted hematopoietic stem cell transplant well before the DLBCL occurrence. This likely would impact the onset and/or management strategies for the cancer.
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2026-04-03 | Abstract 206: Dependencies of diffuse large B-cell lymphoma on EBV oncoproteins.
Abstract Background: Up to 14% of diffuse large B-cell lymphomas (DLBCL) are Epstein-Barr virus-positive (EBV+), and their growth and survival are dependent on EBV latency gene products. DLBCL is a heterogeneous group of aggressive lymphomas, and we previously found that EBV is associated with the BN2 LymphGen subtype. Additionally, multiple genes were mutated at lower frequencies in EBV+ tumors relative to EBV-, suggesting viral oncogenes functionally replace host driver mutations. To identify the EBV oncogenes essential for EBV+ DLBCL and the functional consequences of their inactivation, we are conducting CRISPR/Cas13d screens in EBV+ DLBCL cell lines. Methods: EBV+ DLBCL cell lines (Farage, BCKN1, IBL1, and IBL4) were characterized using whole-exome sequencing (WES), RNA sequencing (RNA-seq), and immunoblotting. We compared the mutational and transcriptomic profiles of EBV+ tumors and cell lines to inform CRISPR/Cas13d functional screens. These screens aim to identify EBV gene product dependencies in EBV+ DLBCL and the impact on cell growth and survival. Results: WES analysis revealed the genetic subtype of each EBV+ DLBCL cell line: Farage was classified as ST2, and the rest were classified as “Other”. RNA-seq and immunoblotting revealed that all expressed an atypical latency III program, characterized by the absence of LMP2A expression. This novel latent gene program was also observed in the EBV+ DLBCL tumors. All cell lines were infected with EBV1, except IBL1, which, to our knowledge, is the only known cell line infected with an EBV1/EBV2 intertypic recombinant. By employing Cas13d for our knockdown screen, we can determine the specific transcripts within the complex EBV latent transcriptome that are essential for the growth and survival of EBV+ DLBCL. Targeting RNA also avoids excessive DNA damage and low sensitivity that arises from targeting 10-20 copies of EBV DNA per cell with Cas9. We have stably expressed two different Cas13d orthologs in each cell line and are optimizing their RNA knockdown efficiency using a FACS-based transient knockdown strategy. Use of clones achieving the most efficient knockdown will allow us to identify essential EBV latent genes and determine the phenotypic consequences (growth arrest, apoptosis, etc.) of inactivating each essential viral oncogene. Conclusions: EBV+ DLBCL exhibits distinct host mutation and gene expression profiles, including the replacement of several subtype-defining mutations with EBV oncogenes. The available EBV+ DLBCL cell lines faithfully model the ST2 or “Other” LymphGen subtypes and exhibit the same atypical EBV latent gene expression pattern as tumors. By determining EBV oncogene dependencies and the effects of their inactivation, we aim to uncover how EBV cooperates with host driver mutations in the pathogenesis of EBV+ DLBCL. Understanding these therapeutic vulnerabilities will provide a foundation for precision therapy. Citation Format: Lily Catherine Foreman Wenger, Quincy Rosemarie, Eric C. Johannsen. Dependencies of diffuse large B-cell lymphoma on EBV oncoproteins [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 206.
2026-03-24 | Same-Slide Spatial Multi-Omics Integration with IN-DEPTH Reveals Tumor Virus-Linked Spatial Reorganization of the Tumor Microenvironment.
Spatial transcriptomics and proteomics have enabled profound insights into tissue organization, yet these technologies remain largely disparate, and emerging same-slide multi-omics approaches are limited in plex, spatial resolution, signal retention, and integrative analytics. We introduce IN-situ DEtailed Phenotyping To High-resolution transcriptomics (IN-DEPTH), a streamlined, resource-efficient, commercially compatible workflow using single-cell spatial proteomics-derived imaging to guide transcriptomic capture on the same slide without RNA signal loss. To integrate modalities beyond niche-level mapping, we developed Spectral Graph Cross-Correlation (SGCC), a proteomic-transcriptomic framework resolving spatially coordinated functional state changes across interacting cell populations. Applied to diffuse large B-cell lymphoma (DLBCL), IN-DEPTH and SGCC enabled stepwise discovery from EBV-positive and EBV-negative tumor comparisons to single-cell resolution, revealing coordinated tumor-macrophage-CD4 T-cell remodeling, immunosuppressive C1Q macrophage enrichment, CD4 T-cell dysfunction, and a candidate IL-27-STAT3 signaling axis. Collectively, IN-DEPTH enables scalable spatial multi-omics to uncover clinically relevant microenvironmental mechanisms and towards robust spatial multi-modal AI models.
2025-11-27 | EBV-Positive Diffuse Large B-Cell Lymphoma (DLBCL): An Assessment and a Roadmap.
Diffuse large B-cell Lymphoma (DLBCL) is the most common lymphoma subtype worldwide. The Epstein Barr Virus (EBV) is a human DNA virus associated with a spectrum of human malignancies. EBV is detected in 5% to 15% of DLBCL tumors with no history of immunosuppression. Early reports of EBV-positive DLBCL suggested distinct clinical presentation, unique tumor microenvironment (TME), and inferior survival compared to EBV-negative DLBCL. A better definition of EBV-positive cases treated with modern chemo-immunotherapy has recently improved our understanding of this entity. Recent studies have provided initial insights into the distinct mutational landscape and immune microenvironment of EBV-positive DLBCL and identified EBV as an independent prognostic biomarker in DLBCL. Despite these advances, fundamental questions remain. We provide an overview of emerging clinical and molecular aspects of EBV-positive DLBCL and discuss recent efforts to develop EBV-targeting therapies. We also discuss key research questions in EBV-positive DLBCL.
2023-06-05 | EBV-positive pyothorax-associated lymphoma expresses CXCL9 and CXCL10 chemokines that attract cytotoxic lymphocytes via CXCR3.
Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma associated with chronic inflammation (DLBCL-CI) develops in the setting of long-standing inflammation. This type of lymphoma may have specific expression profiles of chemokines involved in the pathogenesis of DLBCL-CI. EBV-positive pyothorax-associated lymphoma (PAL) is a prototype of DLBCL-CI and represents a valuable model for the study of this disease category. Using a panel of PAL cell lines, we found that PAL cells expressed and secreted C-X-C motif chemokine ligands 9 and 10 (CXCL9 and CXCL10), the ligands of CXCR3, in contrast to EBV-negative DLBCL cell lines, which did not. Culture supernatants from PAL cell lines attracted CXCR3-expressing CD4+ T cells, CD8+ T cells, and CD56+ natural killer cells from human peripheral blood mononuclear cells. PAL cells injected into mice also attracted CXCR3-positive cytotoxic lymphocytes that expressed interferon-γ. The expression of CXCL9 and CXCL10 was detected in PAL tumor biopsy samples from patients, and CXCR3-positive lymphocytes were abundant in the tissue samples. Collectively, these findings suggest that CXCL9 and CXCL10 are produced by PAL cells and can elicit cytotoxic responses via CXCR3. This chemokine system is also likely to contribute to tissue necrosis, which is a signature histological feature of DLBCL-CI. Further studies are warranted to determine whether the CXCL9-CXCL10/CXCR3 axis exerts antitumor effects in DLBCL-CI.
2020-12-10 | IL-21 Stimulates the expression and activation of cell cycle regulators and promotes cell proliferation in EBV-positive diffuse large B cell lymphoma.
The clinical features of EBV-positive diffuse large B cell lymphoma (DLBCL) indicate a poorer prognosis than EBV-negative DLBCL. Currently, there is no efficacious drug for EBV-positive DLBCL. The cytokine interleukin-21 (IL-21) has been reported to be pro-apoptotic in DLBCL cell lines and is being explored as a new therapeutic strategy for this type of lymphomas. However, our previous studies showed that IL-21 stimulation of EBV-positive DLBCL cell lines leads to increased proliferation. Here, analysis of a rare clinical sample of EBV-positive DLBCL, in combination with a NOD/SCID mouse xenograft model, confirmed the effect of IL-21 on the proliferation of EBV-positive DLBCL cells. Using RNA-sequencing, we identified the pattern of differentially-expressed genes following IL-21 treatment and verified the expression of key genes at the protein level using western blotting. We found that IL-21 upregulates expression of the host MYC and AP-1 (composed of related Jun and Fos family proteins) and STAT3 phosphorylation, as well as expression of the viral LMP-1 protein. These proteins are known to promote the G1/S phase transition to accelerate cell cycle progression. Furthermore, in NOD/SCID mouse xenograft model experiments, we found that IL-21 treatment increases glucose uptake and angiogenesis in EBV-positive DLBCL tumours. Although more samples are needed to validate these observations, our study reconfirms the adverse effects of IL-21 on EBV-positive DLBCL, which has implications for the drug development of DLBCL.
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2026-06-28 | Epstein‐Barr Virus MicroRNAs as Key Regulators of Lymphoma Pathogenesis: Immune Evasion Mechanisms and Therapeutic Opportunities
The ubiquitous human gamma-herpesvirus Epstein-Barr virus (EBV) infects over 90% of adults globally and was the first human virus identified with oncogenic potential. EBV enters a lifelong persistence in the host via a finely regulated life-cycle comprising primary infection, latency and lytic reactivation. Within infected B-cells and epithelial cells, EBV encodes a distinct repertoire of microRNAs (miRNAs), primarily from the BART (BamHI A rightward transcript) and BHRF1 (BamHI H rightward open reading frame) clusters, which play pivotal roles in modulating both viral and host gene expression. These viral miRNAs contribute to key oncogenic processes: by dampening apoptotic responses (e.g., via targeting PUMA, Bim, and PTEN), promoting proliferation of latently-infected B-cells, inhibiting host immune responses (e.g., via down-regulation of CXCL-11 by miR-BHRF1-3), and promoting epithelial-mesenchymal transition (EMT) and metastasis through modulation of E-cadherin and other adhesion molecules. In human lymphomas, such as Burkitt lymphoma, Hodgkin lymphoma, and EBV-positive diffuse large B-cell lymphoma, the interplay of latent viral gene expression, miRNA-mediated regulatory networks, and host microenvironmental factors underlies malignant transformation and disease progression. Emerging evidence also supports the utility of EBV-encoded miRNAs as diagnostic and prognostic biomarkers in EBV-associated cancers. Importantly, therapeutic strategies aimed at interrupting viral miRNA function, restoring host tumor-suppressor pathways, and re-sensitizing tumor cells to immune surveillance hold promise. This review synthesizes current mechanistic insights into EBV-encoded miRNAs in oncogenesis, elaborates on their roles in lymphoma pathogenesis, and evaluates the translational potential of miRNA-targeted therapies in EBV-associated malignancies.
2023-09-08 | Alternative splicing of its 5′-UTR limits CD20 mRNA translation and enables resistance to CD20-directed immunotherapies
Aberrant skipping of coding exons in CD19 and CD22 compromises the response to immunotherapy in B-cell malignancies. Here, we showed that the MS4A1 gene encoding human CD20 also produces several messenger RNA (mRNA) isoforms with distinct 5' untranslated regions. Four variants (V1-4) were detected using RNA sequencing (RNA-seq) at distinct stages of normal B-cell differentiation and B-lymphoid malignancies, with V1 and V3 being the most abundant. During B-cell activation and Epstein-Barr virus infection, redirection of splicing from V1 to V3 coincided with increased CD20 positivity. Similarly, in diffuse large B-cell lymphoma, only V3, but not V1, correlated with CD20 protein levels, suggesting that V1 might be translation-deficient. Indeed, the longer V1 isoform contained upstream open reading frames and a stem-loop structure, which cooperatively inhibited polysome recruitment. By modulating CD20 isoforms with splice-switching morpholino oligomers, we enhanced CD20 expression and anti-CD20 antibody rituximab-mediated cytotoxicity in a panel of B-cell lines. Furthermore, reconstitution of CD20-knockout cells with V3 mRNA led to the recovery of CD20 positivity, whereas V1-reconstituted cells had undetectable levels of CD20 protein. Surprisingly, in vitro CD20-directed chimeric antigen receptor T cells were able to kill both V3- and V1-expressing cells, but the bispecific T-cell engager mosunetuzumab was only effective against V3-expressing cells. To determine whether CD20 splicing is involved in immunotherapy resistance, we performed RNA-seq on 4 postmosunetuzumab follicular lymphoma relapses and discovered that in 2 of them, the downregulation of CD20 was accompanied by a V3-to-V1 shift. Thus, splicing-mediated mechanisms of epitope loss extend to CD20-directed immunotherapies.
2022-03-24 | CircEAF2 counteracts Epstein-Barr virus-positive diffuse large B-cell lymphoma progression via miR-BART19-3p/APC/β-catenin axis.
Epstein-Barr virus (EBV) represents an important pathogenic factor of lymphoma and is significantly associated with poor clinical outcome of diffuse large B-cell lymphoma (DLBCL). Circular RNAs (circRNAs) play an essential role in lymphoma progression. However, the underlying mechanism of circRNA on DLBCL progression related to EBV remains largely unknown. CircRNA was screened by high-throughput sequencing in tumor samples of 12 patients with DLBCL according to EBV infection status. Expression of circEAF2, as well as the relationship with clinical characteristics and prognosis, were further analyzed in tumor samples of 100 DLBCL patients using quantitative real-time PCR. Gain- and loss-of-function experiments were conducted to investigate the biological functions of circEAF2 both in vitro and in vivo. The underlying mechanism of circRNA on DLBCL progression were further determined by RNA sequencing, RNA pull down assay, dual-luciferase reporter assay, rescue experiments and western blotting. We identified a novel circRNA circEAF2, which was downregulated in EBV + DLBCL and negatively correlated with EBV infection and DLBCL progression. In EBV-positive B lymphoma cells, circEAF2 overexpression induced lymphoma cell apoptosis and sensitized lymphoma cells to epirubicin. As mechanism of action, circEAF2 specifically targeted EBV-encoded miR-BART19-3p, upregulated APC, and suppressed downstream β-catenin expression, resulting in inactivation of Wnt signaling pathway and inhibition of EBV + DLBCL cell proliferation. In EBV-positive B-lymphoma murine models, xenografted tumors with circEAF2 overexpression presented decreased Ki-67 positivity, increased cell apoptosis and retarded tumor growth. CircEAF2 counteracted EBV + DLBCL progression via miR-BART19-3p/APC/β-catenin axis, referring circEAF2 as a potential prognostic biomarker. Therapeutic targeting EBV-encoded miRNA may be a promising strategy in treating EBV-associated lymphoid malignancies.
2021-07-08 | Deletion of Viral microRNAs in the Oncogenesis of Epstein–Barr Virus-Associated Lymphoma
Epstein–Barr virus (EBV), which encodes >80 genes and nearly 50 non-coding RNAs, is a double-stranded DNA virus. EBV is associated with various types of lymphomas and lymphoproliferative disorders not only of B-cell but also T/NK-cell origin. However, the oncogenic mechanism remains poorly understood, including the EBV receptors expressed on T/NK cells, relationship of EBV with host genes, and epigenetic regulation of EBV and host genes. The roles of host and viral non-coding RNAs during tumorigenesis have been elucidated. EBV encodes at least 49 mature microRNAs (miRNAs), of which 44 are located in Bam HI-A rightward transcripts (BARTs) region, and the remaining five are located in Bam HI-H rightward fragment 1. BART miRNAs modulate cell differentiation, proliferation, apoptosis, and the cell cycle, and they are considered positive regulators of oncogenesis. We and others have recently reported that EBV-positive lymphomas frequently possess large deletions in BART miRNA clusters, suggesting that some viral miRNAs have suppressive effects on oncogenesis, and that deletion of these miRNAs may aid lymphoma formation.
2020-03-24 | EBV microRNA-BHRF1-2-5p targets the 3'UTR of immune checkpoint ligands PD-L1 and PD-L2.
Epstein-Barr virus-positive (EBV+) diffuse large B-cell lymphomas (DLBCLs) express high levels of programmed death ligand 1 (PD-L1) and PD-L2. MicroRNA (miR) regulation is an important mechanism for the fine-tuning of gene expression via 3'-untranslated region (3'UTR) targeting, and we have previously demonstrated strong EBV miR expression in EBV+ DLBCL. Whereas the EBV latent membrane protein-1 (LMP1) is known to induce PD-L1/L2, a potential counterregulatory role of EBV miR in the fine-tuning of PD-L1/L2 expression remains to be established. To examine this, a novel in vitro model of EBV+ DLBCL was developed, using the viral strain EBV WIL, which unlike common laboratory strains retains intact noncoding regions where several EBV miRs reside. This enabled interrogation of the relationship among EBV latency genes, cell of origin (COO), PD-L1, PD-L2, and EBV miRs. The model successfully recapitulated the full spectrum of B-cell differentiation, with 4 discrete COO phases: early and late germinal center B cells (GCBs) and early and late activated B cells (ABCs). Interestingly, PD-L1/L2 levels increased markedly during transition from late GCB to early ABC phase, after LMP1 upregulation. EBV miR-BamHI fragment H rightward open reading frame 1 (BHRF1)-2-5p clustered apart from other EBV miRs, rising during late GCB phase. Bioinformatic prediction, together with functional validation, confirmed EBV miR-BHRF1-2-5p bound to PD-L1 and PD-L2 3'UTRs to reduce PD-L1/L2 surface protein expression. Results indicate a novel mechanism by which EBV miR-BHRF1-2-5p plays a context-dependent counterregulatory role to fine-tune the expression of the LMP1-driven amplification of these inhibitory checkpoint ligands. Further identification of immune checkpoint-targeting miRs may enable potential novel RNA-based therapies to emerge.
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2026-07-12 | Spatial Architecture of macrophages and T-cells Orchestrates Anti-Lymphoma Immunity and the Response to Immunochemotherapy in EBV+DLBCL
Epstein-Barr Virus (EBV)-positive Diffuse Large B-Cell Lymphoma (EBV+ DLBCL) is a rare and aggressive B-cell lymphoma characterized by heterogeneous pathological features and poor clinical outcomes. The cellular characteristics and spatial architecture of EBV+ DLBCL tumor microenvironment (TME) and its underlying mechanisms remain elusive. Twenty-one EBV+ DLBCL and 24 EBV-negative DLBCL (EBV- DLBCL) lymph node samples were analyzed using imaging mass cytometry (IMC) and the digital spatial profiler (DSP). Our findings revealed that EBV+ DLBCL exhibited a more suppressive TME with increased recruitment of CD163+ and IDO+ tumor-associated macrophages (TAMs) and more exhausted T cells compared to EBV- DLBCL. Consistently, spatial interaction analysis showed that T cells around TAMs displayed more exhausted phenotype. Additionally, more TAMs preemptively hijacked T cells before T-cell mediated attack in EBV+ DLBCL. Mechanistically, more TAMs recruited by EBV+ DLBCL resulted in upregulation of CCL5 through activation of JAK3/STAT3 pathway. Clinically, responders to immunochemotherapy exhibit higher levels of TAMs residing around tumor cells under T-cell attack compared to non-responders. Patients receiving immunomodulatory drugs (PD-1 inhibitors and lenalidomide) combined with immunochemotherapy seem to achieve better outcome. These insights enhance our understanding of the immune architecture in EBV+ DLBCL and identify new targets for immunotherapy, potentially improving patient outcomes.
2026-04-03 | Abstract 7218: Preliminary exploration of the synergistic mechanism between EBV LMP1 and MYD88 L265P mutation on the efficacy of PD-1 Inhibitors in ABC-DLBCL
Abstract Background: Activated B-cell-like diffuse large B-cell lymphoma (ABC-DLBCL) is aggressive with poor prognosis. Epstein-Barr virus (EBV) infection and the MYD88 L265P mutation are key molecular drivers in ABC-DLBCL, typically exhibiting mutual exclusivity, potentially due to functional overlap. PD-1 inhibitors show limited efficacy in unselected relapsed/refractory DLBCL patients. However, our retrospective analysis of relapsed/refractory non-GCB DLBCL patients suggested that rare "double-positive" (EBV-positive/MYD88 L265P mutant) patients had higher response rates to PD-1 inhibitors. Objective: This study investigated how EBV LMP1 modulates intracellular signaling in the context of MYD88 L265P mutation (causing constitutive JAK/STAT activation) and evaluated its synergistic effect with the PD-1 inhibitor tislelizumab. Methods: Clinical data from 54 relapsed/refractory non-GCB DLBCL patients treated with PD-1 antibodies were analyzed. Patients were stratified by EBER ISH (EBV) and MYD88 sequencing. The HBL-1 cell line (MYD88 L265P mutant, EBV-negative) was used. LMP1 was overexpressed via transfection. Groups: control, tislelizumab alone, LMP1 overexpression, LMP1+tislelizumab. qPCR measured mRNA levels of NF-κB p65, MMP9, c-Myc, TLR3, STAT3, JAK3, LMP1, PD-1, PD-L1. Western blot detected protein expression and phosphorylation of key molecules. Results: Among 21 patients with complete molecular data, both "double-positive" patients achieved an objective response (100%), compared to 22.2% in EBV-negative/MYD88 mutant (n=9) and 60.0% in EBV-positive/MYD88 wild-type (n=5) groups. HBL-1 cells showed high baseline p-JAK3 and p-STAT3. LMP1 overexpression suppressed p-JAK3 and p-STAT3. HBL-1 cells expressed tumor cell-intrinsic PD-1 mRNA. Tislelizumab alone inhibited p-JAK3/p-STAT3 phosphorylation. The combination of LMP1 overexpression and tislelizumab showed the strongest inhibition, indicating synergy. Conclusion: MYD88 L265P mutation may cause tumor cell addiction to STAT3 activation maintained by tumor cell-intrinsic PD-1 signaling. LMP1 co-existence adds inhibitory pressure, potentially enhancing this dependency. Tislelizumab blockade may yield a synthetic lethal effect. Combined EBV status and MYD88 profiling could predict PD-1 inhibitor sensitivity in DLBCL, requiring further validation. Citation Format: Chang Wang, Yingtao Lin, Jiesong Wang, . Preliminary exploration of the synergistic mechanism between EBV LMP1 and MYD88 L265P mutation on the efficacy of PD-1 Inhibitors in ABC-DLBCL [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7218.
2026-04-01 | Correction
Article title: Characteristics and prognosis of Epstein Barr virus-positive diffuse large B-cell lymphoma: a retrospective, single-center studyAuthors: Qiaochu, L., Lianming, L., Jianzhen, S., Xiao...
2026-03-07 | Epstein–Barr virus-associated lymphoma: current understanding and treatment strategies
Epstein-Barr virus (EBV) infection is well-known for its high prevalence rate, association with several diseases including cancer and autoimmune conditions, and a wide variety of symptoms and prognosis. When acquired at a young age, primary infections are often asymptomatic; however, in adolescence and young adulthood, symptomatic infections develop, such as in infectious mononucleosis. A special feature of EBV infection is its ability to establish a latent infection in B cells which can lead to long-term infection. Subsequent cellular transformation and viral protein expression can result to EBV-mediated carcinogenesis. Latent proteins expressed by EBV play a role in the pathogenesis of EBV infection and carcinogenesis. These proteins are responsible for a diverse range of functions including cell transformation, cell reprogramming, immune evasion, immune suppression, angiogenesis, cell cycle regulation, and B-cell receptor mimicry.EBV infection is associated with diffuse large B-cell lymphoma, Hodgkin lymphoma, NK/T cell lymphoma, Burkitt lymphoma, post-transplant lymphoproliferative disorders, and primary CNS lymphoma. The clinical presentation varies depending on the specific disease and EBV status, with EBV-positive lymphomas generally associated with poorer prognosis than EBV-negative cases.This review aimed to examine the current understanding of the pathogenesis of EBV-associated lymphoma and to evaluate emerging and accepted therapeutic strategies.
2026-01-09 | Rapidly progressive EBV-positive DLBCL arising from a duodenal diverticulum
Rapidly progressive EBV-positive DLBCL arising from a duodenal diverticulum An 81-year-old man with a history of diabetes mellitus presented with hematemesis.Urgent esophagogastroduodenoscopy (EGD) revealed an adherent blood clot inside a periampullary duodenal diverticulum, suggesting diverticular bleeding ( Fig. 1 A).Since no active bleeding was endoscopically observed, conservative therapy was adopted.Five days later, follow-up endoscopy showed an apparent benign ulcer inside the diverticulum ( Fig. 1 B).Two months later, the patient recurred melena and repeat EGD demonstrated a large ulcerative lesion surrounded by irregular elevations at the same site ( Fig. 1 C).Bioptic examination revealed diffuse infiltration of large atypical lymphoid cells in the stroma.Immunohistochemistry showed CK AE1/AE3(-), CD3(-), CD20( + ), CD5(-), CD10(-), BCL2( + ), BCL6(-), MUM1( + ), and EBER( + ) ( Fig. 2 ), establi-
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2026-07-28 | Clinical-Stage Nanatinostat Triggers Productive Epstein-Barr Virus Lytic Replication.
A phase 1b/2 study of the all-oral "kick-and-kill" combo nanatinostat + valganciclovir (nana-val) showed a ~40% overall response rate in patients with relapsed/refractory Epstein-Barr virus (EBV)-associated lymphomas. Despite the promising results in these difficult-to-treat cancers, there is little known about the effects of nanatinostat on EBV and cell death. In this work we demonstrate that nanatinostat, an HDAC1/3 inhibitor, robustly induces the full EBV lytic cycle in a panel of EBV-positive lymphoma and transformed cell lines (Burkitt, DLBCL, lymphoblastoid), spanning viral latency types and EBV genotypes. Nanatinostat drives lytic-gene transcription across all kinetic classes, viral genome replication, and virion release, and kills cells at nanomolar concentrations. Although valganciclovir does not increase cytotoxicity, its ability to block viral DNA synthesis and virion production justifies its inclusion to prevent viral dissemination. Importantly, the two main metabolites of nanatinostat lack histone-H3 acetylation activity and neither activate nor inhibit the EBV lytic phase, indicating that they are biologically inert with respect to EBV. These mechanistic insights clarify the clinical efficacy of nana-val and support further development of kick-and-kill strategies for EBV-driven malignancies.
2026-05-05 | Adult T-Cell Leukemia/Lymphoma and Epstein-Barr Virus-Positive DLBCL: A Rare Concomitant Association.
Adult T-cell leukemia/lymphoma (ATLL) is an aggressive lymphoma with a poor prognosis. The human T-lymphotropic virus 1 (HTLV-1) is associated with immunodeficiency and increased extranodal involvement in patients with diffuse large B-cell lymphoma (DLBCL). We report on a 47-year-old woman with spastic paraparesis and hepatitis B who was diagnosed with the acute form of ATLL. The clinical picture reveals peripheral generalized lymphadenopathy and splenomegaly. Findings on a hematologic exam indicated leukocytosis with lymphocytosis. A bone marrow biopsy/aspiration confirmed 50% T-cell lymphoid infiltration. Biochemistry results revealed hypercalcemia and a high lactate dehydrogenase value. Results of a CT scan indicated abdominal and thoracic adenopathy as well as moderate splenomegaly. A supraclavicular lymph node biopsy established a DLBCL diagnosis. The final diagnosis was composite lymphoma, DLBCL, and ATLL. The CHOP (cyclophosphamide, doxorubicin hydrochloride, vincristine sulfate [Oncovin], and prednisone) regimen was chosen due to the patient's ECOG performance status. Multiple infectious complications were diagnosed during chemotherapy-induced secondary aplasia. A complete remission, confirmed via PET-CT imaging, was obtained. After 1 month, a skin tumor on the upper right thigh was discovered and biopsied, and the histopathological exam and immunochemistry findings indicated Epstein-Barr virus-DLBCL lymphoma. The association of 2 aggressive lymphomas in a single HTLV-1 carrier is a rare report, and the evolution was severe, complicated by opportunistic infections, and unfavorable.
2026-01-16 | ORY1001 enhances anti-tumor immunity via CXCL10-mediated macrophage modulation in EBV-positive DLBCL.
Epstein-Barr virus-positive diffuse large B-cell lymphoma (EBV+ DLBCL) is an aggressive malignancy characterized by an immunosuppressive tumor microenvironment (TME), where tumor-associated macrophages (TAMs) often exhibit an M2-like phenotype that facilitates immune evasion. Here, we show that the LSD1 inhibitor ORY1001 reprograms the TME of EBV+ DLBCL through CXCL10-mediated modulation of macrophage polarization. Using a reductionist model in which EBV-negative DLBCL cell lines were transfected with LMP1, we demonstrate that ORY1001 induces robust expression of CXCL10, TNF-α and IL-6 in LMP1+ Riva cells, promoting M1 macrophage differentiation and reprogramming M2 macrophages toward a pro-inflammatory phenotype. This CXCL10-driven macrophage reprogramming was also observed in an additional LMP1+ DLBCL line (U2932) and in primary EBV+ DLBCL cells, in which ORY1001 selectively increased CXCL10 levels in tumor-cell supernatants, and conditioned media from these cultures—but not direct ORY1001 exposure—drove M1 polarization of THP-1 and primary CD14+ monocytes. These effects were dependent on CXCL10-CXCR3 signaling, as pharmacological inhibition of CXCR3 abrogated M1 polarization. In vivo, ORY1001 suppressed tumor growth in LMP1-expressing A20 lymphoma models, increased CXCL10 levels, and enhanced M1 macrophage infiltration. Furthermore, ORY1001 synergized with anti–PD-1 therapy to enhance T-cell activation, granzyme B expression, and tumor regression. These findings highlight LSD1 inhibition as a promising immunoepigenetic strategy to remodel the TME and improve the efficacy of immune checkpoint blockade in EBV-associated lymphomas.
2025-10-17 | Withaferin A inhibits EBV-driven lymphomagenesis through multiple mechanisms, including EBNA1 degradation.
Epstein-Barr virus (EBV) infects over 90% of the global population and drives multiple aggressive B-cell malignancies, including Burkitt lymphoma, diffuse large B-cell lymphoma, and Hodgkin lymphoma. Standard chemoimmunotherapy regimens can be highly effective, yet Epstein-Barr virus positive (EBV+) lymphomas sometimes exhibit poorer responses, higher resistance, and worse survival compared with Epstein-Barr virus-negative (EBV-) counterparts. This reflects the virus's ability to drive immune evasion, alter cell death pathways, and exploit host immune dysfunction, underscoring the potential value of EBV-directed strategies. Withaferin A (WA), a steroidal lactone with known anticancer and anti-inflammatory properties, was evaluated for its efficacy against EBV-associated B-cell non-Hodgkin lymphomas (B-NHL). Across a panel of lymphoma cell lines, WA demonstrated selective cytotoxicity toward EBV+ B-NHL, in part through proteasome-dependent degradation of EBNA1 (EBV nuclear antigen 1) and subsequent loss of viral episomes, alongside additional effects on cellular stress and survival pathways. Mechanistic studies revealed that WA collapses antioxidant defenses, drives oxidative stress, and suppresses NF-κB signaling, creating a multipronged disruption of viral and host survival pathways. In primary B-cell models and a cord blood-humanized mouse model of EBV-driven lymphomagenesis, WA inhibited B-cell transformation, reduced splenomegaly and tumor burden, and significantly prolonged survival without evidence of increased viral replication. These findings establish WA as a potent preclinical candidate that selectively targets vulnerabilities unique to EBV-transformed B cells, supporting further optimization and evaluation for EBV+ B-cell malignancies.
2025-09-30 | Immune Checkpoint Molecule Indoleamine 2,3-Dioxygenase 1 (IDO1) Is Expressed in Lymphoma Subtypes With and Without Epstein-Barr Virus.
Indoleamine 2,3-dioxygenase 1 (IDO1) is an immune regulator involved in innate and acquired immunity and immune escape of tumors. Its expression in the tumor microenvironment of several solid tumors and lymphomas associated with Epstein-Barr virus (EBV), together with the promising results from clinical trials of IDO1 inhibitors, prompted us to evaluate IDO1 expression in a large cohort of 455 lymphomas including 154 cases associated with EBV. We optimized an immunohistochemical assay to evaluate IDO1 staining and show that IDO1 expression in seen in several lymphoma subtypes including classic Hodgkin lymphoma (CHL, 40%), diffuse large B-cell lymphoma (DLBCL, 23.5%), lymphoproliferative disorders in post-transplant settings (LPD-PT, 71.4%), extranodal NK/T-cell lymphoma (ENKTL, 92%), and ALK-negative anaplastic large cell lymphoma (ALCL, 39%), among others. Multiplex immunofluorescence further aided in refining the localization of IDO1 protein expression particularly within the tumor microenvironment. There was a significant correlation between IDO1 expression and EBV positivity in CHL (83.8%), LPD-PT (86.2%), and ENKTL (91.5%), with statistically significant difference in the mean IDO1 H-scores between EBV-positive and EBV-negative cases. IDO1 expression in ALCL was confined to ALK-negative cases with a significant correlation between IDO1 expression and ALK status. Our findings show that IDO1 expression is not only highly correlated with lymphomas associated with EBV, but also found in lymphomas unassociated with EBV, including aggressive and refractory subtypes of lymphomas for which immune checkpoint inhibition through IDO1 could be exploited for therapeutic purposes.
cell therapies
2025-12-01 | Successful Use of Haploidentical HSCT in a Child With Schimke Immuno-Osseous Dysplasia Who Developed PTLD After Kidney Transplantation.
Schimke Immuno-Osseous Dysplasia Is a Rare Autosomal Recessive Multisystem Disorder Caused by Biallelic Pathogenic Variants in the SMARCAL1 Gene, Which Encodes a DNA Annealing Helicase Essential for Replication Fork Stability and Genomic Maintenance. Loss of SMARCAL1 Function Leads to Genomic Instability, Resulting in a Characteristic Clinical Triad of Disproportionate Short Stature, Steroid-Resistant Nephrotic Syndrome and Immunodeficiency. Kidney Transplantation Is the Standard Treatment for End-Stage Renal Disease in Schimke Immuno-Osseous Dysplasia. However, the Underlying Genomic Fragility and Immunodeficiency Heighten the Risk of Post-Transplant Complications, Particularly Epstein-Barr Virus-Associated Post-Transplant Lymphoproliferative Disorder. We Report the Case of a 7-Year-Old Girl With Genetically Confirmed Schimke Immuno-Osseous Dysplasia Who Developed Epstein-Barr Virus-Positive Diffuse Large B-Cell Lymphoma Eight Months Following Maternal Kidney Transplantation, Despite Reduced-Intensity Immunosuppression. Rituximab Therapy Resulted in Complete Remission, but Required the Withdrawal of all Immunosuppressive Agents, Thereby Posing a Risk of Kidney Graft Loss. In the Context of Primary Immunodeficiency and the Development of Post-Transplant Lymphoproliferative Disorder in Such a Setting, the Patient Subsequently Underwent Haploidentical Hematopoietic Stem Cell Transplantation From the Mother, Using an αβ T-Cell-Depleted Graft and a Nephrotoxicity-Sparing Conditioning Regimen. Full Donor Chimerism Was Achieved by Day 28. Immune Reconstitution Occurred Within One Year. At 27 Months After Hematopoietic Stem Cell Transplantation She Remains in Remission With Preserved Kidney Function and no Ongoing Immunosuppression. This Case Highlights the Feasibility of Haploidentical Hematopoietic Stem Cell Transplantation as a Curative Salvage Strategy for Immunosuppression-Free Survival After Kidney Transplantation in Patients With Schimke Immuno-Osseous Dysplasia.
2025-08-01 | 28P Pre-clinical development of CAR-T cells targeting a lytic EBV antigen gp350 for therapy of EBV-positive diffuse large B-cell lymphoma
Epstein-Barr virus (EBV) associated diffuse large B-cell lymphoma (DLBCL) represents a rare aggressive B-cell lymphoma subtype characterized by an adverse clinical outcome. Despite intensive chemotherapy, the prognosis of EBV-positive DLBCL patients remains unfavorable, and a definitive method to completely eliminate latent EBV infection is lacking. Developing chimeric antigen receptor (CAR)-T cells therapy targeting EBV specific protein in the context of this EBV+ aggressive lymphoma will provide a novel therapeutic strategy for patients with limited treatment options.
2025-06-01 | 686 | TABELECLEUCEL IN EBV POSITIVE POST‐TRANSPLANT LYMPHOPROLIFERATIVE DISEASE: A REAL WORD EXPERIENCE
Introduction: ALLELE trial reported promising data on tabelecleucel, an on-demand allogeneic cytotoxic T-cell therapy for relapse/resistant epstein-barr positive post-transplant lymphoproliferative disease (EBV+ PTLD) that occurred after either solid organ (SOT) or hematopoietic transplantation (HCT), with a median OS improved from less than 6 to more than 18 months. The aim of this study is to evaluate feasibility and clinical outcome of PTLD in our center since the availability of tabelecleucel as compassionate use. Methods: Patients who developed histologically documented PTLD in adulthood after either SOT or HCT were analyzed since November 2023. Results: Between November 2023 and February 2025 we diagnosed 12 consecutive cases of PTLD, 10 after SOT (5 heart, 2 lungs, 1 kidney, 1 liver and 1 multivisceral transplant) and 2 after HCT. Median age at presentation of PTLD was 53 years (range 25–76), 8 patients were male. At diagnosis main adverse clinical features were: stage III–IV (n = 10, 83%), B symptoms (n = 3; 25%); ECOG > 2 (n = 7, 58%), bulky disease (2 patients, 16%). According to IPI score 50% of patients were identified as intermediate-high/high risk. Median time from transplant to PTLD was 86 months (range 3 to 372). We documented 9 DLBCL, 1 PEL, 1 polymorphic lymphoproliferative disorder and 1 primitive CNS lymphoma. EBV-DNA integration in lymphoma cells (EBER +) was documented in 9 patients (75%). All patients received Rituximab monotherapy: 2 patients achieve CR (16%) and did not require other additional therapy. Tabelecleucel was required as compassionate use in Rituximab Resistant EBV+ PTLD in 7 patients (2 after HCT and 5 not eligible/refusing chemotherapy program after SOT). In a median time of 14 days we were able to define the origin of the disease in 6 out of 7 cases (5 from patient, 1 from donor). In less than a week we identified a suitable batch of tabelecleucel in all the cases. One patient died before tabelecleucel delivery due to disease progression. Two patients are planned to be reinfused. For the remaining, median time from request to first Tabelecleucel dose was 22 days (range 11–44). Three patients completed tabelecleucel therapy, one is still under treatment. No hematological or extra hematological toxicities were documented. We documented one CR, one SD and one PD. This latter patient received R-CHOP achieving CR. After a median follow up of 8 months all the three patients are alive. The 3 patients with EBV negative Rituximab refractory PTLDs were treated with R-CHOP obtaining CR and are alive after a median follow up of 7 months. Conclusion: Our single center experience showed that tabelecleucel therapy is feasible in a real word setting. Time from “brain to vein” is short, thus preventing disease progression in the majority of cases. In our experience tabelectelucel can represent a good alternative to chemotherapy in EBV positive Rituximab resistant/refractory lymphoproliferative disorders. Keywords: cellular therapies Potential sources of conflict of interest: A. M. Barbui Consultant or advisory role: Pierre Fabre Honoraria: Pierre Fabre Educational grants: Pierre Fabre
2024-08-21 | Transition From Epstein-Barr Virus (EBV)-Positive Rectal Hodgkin Lymphoma to Diffuse Large B-Cell Lymphoma in the Lung.
We report a distinctive case of sequential lymphomas in a 72-year-old male, initially diagnosed with Epstein-Barr virus (EBV)-positive rectal classic Hodgkin lymphoma (cHL), followed by the development of diffuse large B cell lymphoma (DLBCL) in the lung. This rare progression underscores the complexity of lymphomas associated with EBV infection and their unpredictable clinical courses. The patient's journey began with symptoms of intractable diarrhea, low appetite, and significant weight loss, leading to the diagnosis of stage 4B cHL, managed initially with brentuximab/doxorubicin, vinblastine, dacarbazine (AVD) chemotherapy. Despite a partial response, surveillance identified a transition to DLBCL, marked by new pulmonary lesions. This case highlights the clinical and diagnostic challenges in managing sequential lymphomas, emphasizing the role of EBV in lymphomagenesis and the potential for clonal evolution from a common precursor cell. The therapeutic approach evolved from targeted chemotherapy to consideration of advanced treatments such as autologous stem cell transplant and chimeric antigen receptor (CAR) T-cell therapy, reflecting the aggressive nature and poor prognosis of the disease. This case contributes to our understanding of the EBV's impact on lymphoma progression and underscores the need for vigilant monitoring and adaptive treatment strategies in similar clinical scenarios.
2024-07-19 | Infant with diffuse large B-cell lymphoma identified postmortem with homozygous founder Slavic RAG1 variant: a case report and literature review.
There is an increased risk of lymphomas in inborn errors of immunity (IEI); however, germline genetic testing is rarely used in oncological patients, even in those with early onset of cancer. Our study focuses on a child with a recombination-activating gene 1 (RAG1) deficiency who was identified through a screening program for Slavic founder genetic variants among patients who died with malignancy at an early age in Belarus. We identified one homozygous founder RAG1 variant out of 24 available DNA samples from 71 patients who developed lymphoma aged <3 years from the Belarusian cancer registry between 1986 and 2023. Our patient had an episode of pneumonia at 3 months of age and was hospitalized for respiratory distress, candida-positive lung disease, and lymphadenopathy at 14 months of age. The diagnosis of Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) was established. The patient had a normal lymphocyte count that decreased over time. One month after chemotherapy initiation, the patient died due to sepsis and multiple organ failure without a genetic diagnosis. In a retrospective analysis, T-cell receptor excision circles (TRECs) and kappa-deleting recombination excision circles (KRECs) were undetectable in peripheral blood. A targeted screening program designed to detect a Slavic founder variant in the RAG1 gene among children revealed a 14-month-old Belarusian male infant with low TREC levels who died of EBV-driven DLBCL and complications of chemotherapy including infections. This case highlights how patients with IEI and recurrent infections may develop serious non-infectious complications, such as fatal malignancy. It also emphasizes the importance of early identification, such as newborn screening for severe combined immune deficiency. Earlier diagnosis of RAG deficiency could have prompted hematopoietic stem cell transplant well before the DLBCL occurrence. This likely would impact the onset and/or management strategies for the cancer.
other
2026-04-03 | Abstract 206: Dependencies of diffuse large B-cell lymphoma on EBV oncoproteins.
Abstract Background: Up to 14% of diffuse large B-cell lymphomas (DLBCL) are Epstein-Barr virus-positive (EBV+), and their growth and survival are dependent on EBV latency gene products. DLBCL is a heterogeneous group of aggressive lymphomas, and we previously found that EBV is associated with the BN2 LymphGen subtype. Additionally, multiple genes were mutated at lower frequencies in EBV+ tumors relative to EBV-, suggesting viral oncogenes functionally replace host driver mutations. To identify the EBV oncogenes essential for EBV+ DLBCL and the functional consequences of their inactivation, we are conducting CRISPR/Cas13d screens in EBV+ DLBCL cell lines. Methods: EBV+ DLBCL cell lines (Farage, BCKN1, IBL1, and IBL4) were characterized using whole-exome sequencing (WES), RNA sequencing (RNA-seq), and immunoblotting. We compared the mutational and transcriptomic profiles of EBV+ tumors and cell lines to inform CRISPR/Cas13d functional screens. These screens aim to identify EBV gene product dependencies in EBV+ DLBCL and the impact on cell growth and survival. Results: WES analysis revealed the genetic subtype of each EBV+ DLBCL cell line: Farage was classified as ST2, and the rest were classified as “Other”. RNA-seq and immunoblotting revealed that all expressed an atypical latency III program, characterized by the absence of LMP2A expression. This novel latent gene program was also observed in the EBV+ DLBCL tumors. All cell lines were infected with EBV1, except IBL1, which, to our knowledge, is the only known cell line infected with an EBV1/EBV2 intertypic recombinant. By employing Cas13d for our knockdown screen, we can determine the specific transcripts within the complex EBV latent transcriptome that are essential for the growth and survival of EBV+ DLBCL. Targeting RNA also avoids excessive DNA damage and low sensitivity that arises from targeting 10-20 copies of EBV DNA per cell with Cas9. We have stably expressed two different Cas13d orthologs in each cell line and are optimizing their RNA knockdown efficiency using a FACS-based transient knockdown strategy. Use of clones achieving the most efficient knockdown will allow us to identify essential EBV latent genes and determine the phenotypic consequences (growth arrest, apoptosis, etc.) of inactivating each essential viral oncogene. Conclusions: EBV+ DLBCL exhibits distinct host mutation and gene expression profiles, including the replacement of several subtype-defining mutations with EBV oncogenes. The available EBV+ DLBCL cell lines faithfully model the ST2 or “Other” LymphGen subtypes and exhibit the same atypical EBV latent gene expression pattern as tumors. By determining EBV oncogene dependencies and the effects of their inactivation, we aim to uncover how EBV cooperates with host driver mutations in the pathogenesis of EBV+ DLBCL. Understanding these therapeutic vulnerabilities will provide a foundation for precision therapy. Citation Format: Lily Catherine Foreman Wenger, Quincy Rosemarie, Eric C. Johannsen. Dependencies of diffuse large B-cell lymphoma on EBV oncoproteins [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 206.
2026-03-24 | Same-Slide Spatial Multi-Omics Integration with IN-DEPTH Reveals Tumor Virus-Linked Spatial Reorganization of the Tumor Microenvironment.
Spatial transcriptomics and proteomics have enabled profound insights into tissue organization, yet these technologies remain largely disparate, and emerging same-slide multi-omics approaches are limited in plex, spatial resolution, signal retention, and integrative analytics. We introduce IN-situ DEtailed Phenotyping To High-resolution transcriptomics (IN-DEPTH), a streamlined, resource-efficient, commercially compatible workflow using single-cell spatial proteomics-derived imaging to guide transcriptomic capture on the same slide without RNA signal loss. To integrate modalities beyond niche-level mapping, we developed Spectral Graph Cross-Correlation (SGCC), a proteomic-transcriptomic framework resolving spatially coordinated functional state changes across interacting cell populations. Applied to diffuse large B-cell lymphoma (DLBCL), IN-DEPTH and SGCC enabled stepwise discovery from EBV-positive and EBV-negative tumor comparisons to single-cell resolution, revealing coordinated tumor-macrophage-CD4 T-cell remodeling, immunosuppressive C1Q macrophage enrichment, CD4 T-cell dysfunction, and a candidate IL-27-STAT3 signaling axis. Collectively, IN-DEPTH enables scalable spatial multi-omics to uncover clinically relevant microenvironmental mechanisms and towards robust spatial multi-modal AI models.
2025-11-27 | EBV-Positive Diffuse Large B-Cell Lymphoma (DLBCL): An Assessment and a Roadmap.
Diffuse large B-cell Lymphoma (DLBCL) is the most common lymphoma subtype worldwide. The Epstein Barr Virus (EBV) is a human DNA virus associated with a spectrum of human malignancies. EBV is detected in 5% to 15% of DLBCL tumors with no history of immunosuppression. Early reports of EBV-positive DLBCL suggested distinct clinical presentation, unique tumor microenvironment (TME), and inferior survival compared to EBV-negative DLBCL. A better definition of EBV-positive cases treated with modern chemo-immunotherapy has recently improved our understanding of this entity. Recent studies have provided initial insights into the distinct mutational landscape and immune microenvironment of EBV-positive DLBCL and identified EBV as an independent prognostic biomarker in DLBCL. Despite these advances, fundamental questions remain. We provide an overview of emerging clinical and molecular aspects of EBV-positive DLBCL and discuss recent efforts to develop EBV-targeting therapies. We also discuss key research questions in EBV-positive DLBCL.
2023-06-05 | EBV-positive pyothorax-associated lymphoma expresses CXCL9 and CXCL10 chemokines that attract cytotoxic lymphocytes via CXCR3.
Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma associated with chronic inflammation (DLBCL-CI) develops in the setting of long-standing inflammation. This type of lymphoma may have specific expression profiles of chemokines involved in the pathogenesis of DLBCL-CI. EBV-positive pyothorax-associated lymphoma (PAL) is a prototype of DLBCL-CI and represents a valuable model for the study of this disease category. Using a panel of PAL cell lines, we found that PAL cells expressed and secreted C-X-C motif chemokine ligands 9 and 10 (CXCL9 and CXCL10), the ligands of CXCR3, in contrast to EBV-negative DLBCL cell lines, which did not. Culture supernatants from PAL cell lines attracted CXCR3-expressing CD4+ T cells, CD8+ T cells, and CD56+ natural killer cells from human peripheral blood mononuclear cells. PAL cells injected into mice also attracted CXCR3-positive cytotoxic lymphocytes that expressed interferon-γ. The expression of CXCL9 and CXCL10 was detected in PAL tumor biopsy samples from patients, and CXCR3-positive lymphocytes were abundant in the tissue samples. Collectively, these findings suggest that CXCL9 and CXCL10 are produced by PAL cells and can elicit cytotoxic responses via CXCR3. This chemokine system is also likely to contribute to tissue necrosis, which is a signature histological feature of DLBCL-CI. Further studies are warranted to determine whether the CXCL9-CXCL10/CXCR3 axis exerts antitumor effects in DLBCL-CI.
2020-12-10 | IL-21 Stimulates the expression and activation of cell cycle regulators and promotes cell proliferation in EBV-positive diffuse large B cell lymphoma.
The clinical features of EBV-positive diffuse large B cell lymphoma (DLBCL) indicate a poorer prognosis than EBV-negative DLBCL. Currently, there is no efficacious drug for EBV-positive DLBCL. The cytokine interleukin-21 (IL-21) has been reported to be pro-apoptotic in DLBCL cell lines and is being explored as a new therapeutic strategy for this type of lymphomas. However, our previous studies showed that IL-21 stimulation of EBV-positive DLBCL cell lines leads to increased proliferation. Here, analysis of a rare clinical sample of EBV-positive DLBCL, in combination with a NOD/SCID mouse xenograft model, confirmed the effect of IL-21 on the proliferation of EBV-positive DLBCL cells. Using RNA-sequencing, we identified the pattern of differentially-expressed genes following IL-21 treatment and verified the expression of key genes at the protein level using western blotting. We found that IL-21 upregulates expression of the host MYC and AP-1 (composed of related Jun and Fos family proteins) and STAT3 phosphorylation, as well as expression of the viral LMP-1 protein. These proteins are known to promote the G1/S phase transition to accelerate cell cycle progression. Furthermore, in NOD/SCID mouse xenograft model experiments, we found that IL-21 treatment increases glucose uptake and angiogenesis in EBV-positive DLBCL tumours. Although more samples are needed to validate these observations, our study reconfirms the adverse effects of IL-21 on EBV-positive DLBCL, which has implications for the drug development of DLBCL.
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Drug Discovery Landscape
2 orphan drug designations for Epstein-Barr virus-positive diffuse large B-cell lymphoma.
2 orphan drug designations for Epstein-Barr virus-positive diffuse large B-cell lymphoma.
Drug | Therapy type | Regulator | Orphan designation | Approval | Sponsor |
|---|---|---|---|---|---|
nanatinostat and valganciclovir | small molecules | FDA | 2021-11-24 | — | Viracta Subsidiary, Inc. |
autologous Epstein-Barr virus specific T-cells | cell therapies | FDA | 2015-03-09 | — | BCM Center for Cell and Gene Therapy |
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