Active Epstein-Barr Virus Infection Research Articles

223 - Clinical Experience of Tabelecleucel in Patients with Life-Threatening Complications of Epstein–Barr Virus Viremia

Background Epstein-Barr virus (EBV) viremia - a condition of persistent active (vs latent) EBV infection - can be associated with a variety of life-threatening disorders. One such disorder is chronic active EBV (CAEBV), a type of primary immunodeficiency associated with high mortality rates. Another entity is EBV-driven (EBV+) hemophagocytic lymphohistiocytosis (HLH), characterized by a hyperactive immune response with elevated cytokines. Both can be associated with multi-organ dysfunction and failure. In addition, EBV viremia may be associated with complications following allogeneic hematopoietic cell transplantation (HCT) or can herald relapse of EBV+ malignancies. Tabelecleucel, an investigational off-the-shelf, allogeneic EBV-specific T-cell immunotherapy, has previously shown clinical activity in patients with EBV+ post-transplant lymphoproliferative disease (PTLD), EBV+ PTLD with central nervous system involvement and EBV+ leiomyosarcoma (Prockop S et al, JCI 2020; Kurlander et al, ESMO 2018) and is a potential therapy for CAEBV, HLH or EBVviremia in other life-threatening settings since it may control the EBV-based stimuli underlying these distinct diseases (Icheva V et al, J Clin Oncol 2013). Here, we report data in patients with complications of EBV viremia treated with tabelecleucel in an expanded access study (NCT02822495) within two non-overlapping protocols (EAP-201 [2016-2018] and EAP-901 [2018-present]). Methods Eligible patients received tabelecleucel at ~2 x 106 cells/kg/dose on Days 1, 8 and 15, with laboratory assessments on Day ~28 of each 5-week cycle. EAP-201 enrolled patients with persistent EBV viremia (defined as serum EBV DNA assays showing detectable EBV viral load at least 4 weeks apart and known or suspected immunodeficiency or history of prior EBV+ malignancy) as well as patients with EBV+ HLH. EAP-901 includes patients with persistent EBV viremia and known or suspected immunodeficiency. Patients who did not respond could switch to tabelecleucel with a different human leukocyte antigen (HLA) restriction (restriction switch). In EAP-201, response criteria were based on reduction in EBV viral load and/or on HLH response criteria (Table 1). As EAP-901 is intended to provide compassionate access to tabelecleucel, limited patient data are collected compared with EAP-201; however, response is assessed by the investigators based on reduction in EBV viral load to determine whether an HLA restriction switch is needed. In both protocols, patients continued treatment until unacceptable toxicity, maximal response (2 consecutive complete responses [CR] or 3 partial responses [PR]), or up to 4 different HLA restrictions. Both protocols collected serious AEs (SAEs). Results Patient characteristics, treatment exposure, and responses are reported in Table 1; 5 patients with EBV viremia participated in EAP-201 and 4 in EAP-901. Objective response rates were 80% (n=4/5) for EAP-201 and 50% (n=2/4) for EAP-901. For EAP-201, the median time to first response for the 4 patients who responded was 1.7 (min 1.0, max 4.7) months, and for EAP-901 it was approximately 1 month for both patients who responded. Table 2 summarizes the treatment-emergent SAEs (TESAEs) observed with both protocols. EAP-901 reported a fatal SAE of chronic hepatic failure that was assessed as unrelated to treatment. One patient in EAP-901 experienced a grade 3 TESAE of facial nerve disorder that was assessed as possibly related to tabelecleucel; there were no other fatal or treatment-related TESAEs reported in either protocol, and no safety concern has emerged for this patient population. Conclusions Tabelecleucel was well tolerated and showed evidence of clinical activity in patients with life-threatening EBV viremia. Based on these results, further clinical investigation of tabelecleucel in patients with HLH or CAEBV is planned within an upcoming study (ATA129-EBV-205). Download : Download high-res image (743KB) Download : Download full-size image Disclosures Ghobadi: Amgen: Consultancy, Research Funding. Stiff: CRISPR: Consultancy; Gamida-Cell: Research Funding; Atara Biotherapeutics: Research Funding; Amgen: Research Funding; Incyte: Research Funding; Takeda: Research Funding; Macrogenics: Research Funding; Eisai: Research Funding. Reshef: Kite: Consultancy, Research Funding; Atara Biotherapeutics: Consultancy, Research Funding; Bristol-Myers Squibb: Consultancy; Magenta: Consultancy; Incyte: Consultancy, Research Funding; Pharmacyclics: Consultancy, Research Funding; Pfizer: Consultancy; Celgene: Research Funding; Shire: Research Funding. Navarro: Atara Biotherapeutics: Current Employment, Current equity holder in publicly-traded company. Gamelin: Atara Biotherapeutics: Current Employment, Current equity holder in publicly-traded company. Dinavahi: Atara Biotherapeutics: Current Employment, Current equity holder in publicly-traded company. Sun: Atara Biotherapeutics: Current Employment, Current equity holder in publicly-traded company. Guzman-Becerra: Atara Biotherapeutics: Current Employment, Current equity holder in publicly-traded company. Prockop: Mesoblast: Consultancy, Research Funding; Atara Biotherapeutics: Research Funding; Jasper Pharmaceuticals: Research Funding; Memorial Sloan Kettering: Patents & Royalties: IP related to the development of third party viral specific T cells with all of my interests assigned to MSK.

Chronic Active Epstein-Barr Virus Infection: The Elucidation of the Pathophysiology and the Development of Therapeutic Methods.

Chronic active Epstein–Barr virus infection (CAEBV) is a disease where Epstein–Barr virus (EBV)-infected T- or NK-cells are activated and proliferate clonally. The symptoms of this dual-faced disease include systemic inflammation and multiple organ failures caused by the invasion of infected cells: inflammation and neoplasm. At present, the only effective treatment strategy to eradicate EBV-infected cells is allogeneic stem cell transplantation. Lately, the investigation into the disease’s pathogenic mechanism and pathophysiology has been advancing. In this review, I will evaluate the new definition in the 2017 WHO classification, present the advancements in the study of CAEBV, and unfold the future direction.

The road to treating chronic active Epstein-Barr viral infection

Chronic active Epstein-Barr virus (CAEBV) infection is a progressive disease characterized by persistent inflammatory symptoms accompanied by clonally proliferating EBV-positive T or NK cells. The optimal medical treatment for CAEBV to eradicate EBV-infected T or NK cells has not yet been established, with allogeneic hematopoietic stem cell transplantation as the only strategy currently available. Patients with CAEBV have been reported mainly from limited area of Japan and East Asia. However, CAEBV is drawing a global attention, and the number of reports is increasing worldwide after its definition was added to the EBV-positive T- or NK-cell neoplasms in the 2017 World Health Organization classification. We had previously discovered that STAT3 was constitutively activated in EBV-infected tumor cells in CAEBV inducing the immortalization and production of inflammatory cytokines. Based on these findings, an investigator-initiated clinical research of a JAK1/2 inhibitor ruxolitinib for CAEBV infection was initiated in January 2019. Japanese researchers have been expected to elucidate pathological mechanisms and to establish an effective treatment.

Spontaneous regression of chronic epstein –Barr virus infection-related lymphoproliferative disease

Chronic active Epstein–Barr virus infection (CAEBV) is relatively uncommon and can be associated with lymphoproliferative diseases (LPD's) of NK/T cell and B cell type. In the western world, the disease is less common, while it is found to be more prevalent in the East Asian, South, and Central American population. The presentation widely varies from skin rashes, fever, lymphadenopathy to major organ involvement and usually takes a fulminant course. Here, we describe a 70-year-old Indian male, who was diagnosed as CAEBV-related LPD with spontaneous regression with a brief review of the literature.

Pathology of macrophage activation syndrome in humanized NSGS mice

“Humanized” immunodeficient mice generated via the transplantation of CD34+ human hematopoietic stem cells (hHSC) are an important preclinical model system. The triple transgenic NOD.Cg-PrkdcscidIl2rgtm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ (NSGS) mouse line is increasingly used as recipient for CD34+ hHSC engraftment. NSGS mice combine the features of the highly immunodeficient NSG mice with transgenic expression of the human myeloid stimulatory cytokines GM-CSF, IL-3, and Kit ligand. While generating humanized NSGS (huNSGS) mice from two independent cohorts, we encountered a fatal macrophage activation syndrome (MAS)-like phenotype resulting from the transplantation of CD34+ hHSC. huNSGS mice exhibiting this phenotype declined clinically starting at approximately 10 weeks following CD34+ hHSC engraftment, with all mice requiring euthanasia by 16 weeks. Gross changes comprised small, irregular liver, splenomegaly, cardiomegaly, and generalized pallor. Hematological abnormalities included severe thrombocytopenia and anemia. Pathologically, huNSGS spontaneously developed a disseminated histiocytosis with infiltrates of activated macrophages and hemophagocytosis predominantly affecting the liver, spleen, bone marrow, and pancreas. The infiltrates were chimeric with a mixture of human and mouse macrophages. Immunohistochemistry suggested activation of the inflammasome in both human and murine macrophages. Active Epstein-Barr virus infection was not a feature. Although the affected mice exhibited robust chimerism of the spleen and bone marrow, the phenotype often developed in the face of low chimerism of the peripheral blood. Given the high penetrance and early lethality associated with the MAS-like phenotype here described, we urge caution when considering the use of huNSGS mice for the development of long-term studies.

CD8dimCD3+ lymphocytes in fever patients might be biomarkers of active EBV infection and exclusion indicator of T-LGLL.

Background: Massive monoclonal or oligoclonal expansion of CD8+ T cells is a notable feature of primary infections of the Epstein-Barr virus (EBV). However, the clinical significance of this expansion is not clear. Results: An increase in the CD8dimCD3+ lymphocyte subset in patients with active EBV infection was due to caspase-8-dependent apoptosis was found using flow cytometry in this study. The number of these cells was associated with the illness severity. Pan-T-cell antigen and receptor analyses were also compared in patients with active EBV infections and T-cell large granular lymphocytic leukemia to provide additional diagnostic information. Conclusion: The increase in CD8dimCD3+ cells could be a biomarker of active EBV infection and an exclusion indicator of T-cell large granular lymphocytic leukemia with flow cytometric analysis.

Chronic Active Epstein-Barr Virus Infection: Is It Immunodeficiency, Malignancy, or Both?

Simple SummaryChronic active Epstein–Barr virus (EBV) infection (CAEBV) is a rare syndrome of unknown etiology characterized by prolonged infectious mononucleosis-like symptoms and proliferation of EBV-infected T and/or natural killer cells. CAEBV has been primarily reported in East Asia and Latin America, suggesting a genetic predisposition in its pathogenesis. The clinical course of CAEBV is heterogeneous ranging from an indolent and occasionally self-limiting disease to an aggressive and fatal condition, but its prognosis is generally poor. This heterogeneous clinical picture does not suggest a simple etiology for the syndrome. Clinicopathological investigations of CAEBV suggest that it has aspects of both malignant neoplasm and immunodeficiency. This article summarizes the latest findings on CAEBV and discusses critical unsolved questions regarding its pathogenesis and disease concept.Chronic active Epstein–Barr virus (EBV) infection (CAEBV) is a rare syndrome characterized by prolonged infectious mononucleosis-like symptoms and elevated peripheral blood EBV DNA load in apparently immunocompetent persons. CAEBV has been primarily reported in East Asia and Latin America, suggesting a genetic predisposition in its pathogenesis. In most cases of CAEBV, EBV induces proliferation of its unusual host cells, T or natural killer (NK) cells. The clinical course of CAEBV is heterogeneous; some patients show an indolent course, remaining in a stable condition for years, whereas others show an aggressive course with a fatal outcome due to hemophagocytic lymphohistiocytosis, multiple organ failure, or progression to leukemia/lymphoma. The pathogenesis of CAEBV is unclear and clinicopathological investigations suggest that it has aspects of both malignant neoplasm and immunodeficiency. Recent genetic analyses of both viral and host genomes in CAEBV patients have led to discoveries that are improving our understanding of the nature of this syndrome. This article summarizes the latest findings on CAEBV and discusses critical unsolved questions regarding its pathogenesis and disease concept.

COMPLAINTS OF THE PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND ACTIVE CYTOMEGALOVIRUS AND EPSTEIN-BARR VIRUS INFECTION; THEIR DIAGNOSTIC VALUE

COMPLAINTS OF THE PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND ACTIVE CYTOMEGALOVIRUS AND EPSTEIN-BARR VIRUS INFECTION; THEIR DIAGNOSTIC VALUE

Clinical Characteristics of Hemophagocytic Lymphohistiocytosis Associated with Non-Hodgkin B-Cell Lymphoma: A Multicenter Retrospective Study

BackgroundHemophagocytic lymphohistiocytosis (HLH) associated with B-cell lymphoma is a highly aggressive disease with unclear clinical features and has no standard treatment. Patients and MethodsWe analyzed the clinical characteristics of 31 patients from two individual centers. ResultsThe median overall survival was only 1.5 months. Both univariate and multivariate analyses, based on lymphoma or HLH-related characteristics, revealed that patients with high Epstein-Barr virus (EBV) DNA load and ≥ 2 extranodal lesions, or hypofibrinogenemia, respectively, showed significantly poorer overall survival. Interestingly, some patients with high EBV DNA load had EBV-positive natural killer (NK) and/or T cells, which may be related to the coexistence of immunodeficiency and/or chronic active EBV infection. Molecular genetics examination confirmed that 47.4% (9/19) of patients had complex karyotypes, 37.5% (3/8) of patients had TP53 deletions, and 21.34% (3/14) of patients had TP53 mutation or alteration of malignancy-related pathways, including BCR/NF-κB, JAK-STAT, and epigenetic regulatory pathways, which may provide clues to choose targets for therapy. Treatment regimens containing etoposide, anti-CD20 monoclonal antibodies, or anthracyclines improved patient prognosis (P = .0183, .025, and .0436, respectively). Patients with infections had significantly shorter survival than those without infections (P = .00019). ConclusionThe patients’ performance status, number of extranodal lesions, high EBV DNA load, and hypofibrinogenemia are poor prognostic factors for HLH associated with B-cell lymphoma. Molecular genetic high-risk factors are of particular importance because these factors can provide information for prognosis prediction, treatment decisions, and disease surveillance.

CYTOMEGALOVIRUS AND VIRUS EPSTEIN- BARR INFECTION IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND ITS DEPENDENCE ON GENDER AND AGE OF PATIENTS

Introduction. Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by various manifestations and clinical course, many aspects of the etiology and pathogenesis of which remain unclear. Recently, the interest of researchers in studying the role of cytomegalovirus (CMV) and Epstein - Barr virus (EBV) has been growing in the occurrence and course of a number of human diseases due to their ability to affect almost all organs and systems of the body, causing the formation of latent, active or chronic infection, which can often cause temporary disability, disability or even death, however, for the patients with SLE, despite the possibility of approaching the difficult problem of diagnosis and treatment of this disease, this issue is given insufficient attention, as evidenced by isolated studies.The aim of the study. Detect cytomegalovirus and Epstein - Barr infection in patients with systemic lupus erythematosus and its dependence on gender and age of patients. Materials and methods of research. The study involved 120 patients (15 men (12.50%) and 105 women (87.50%) aged 18 to 69 years with SLE, who were in the rheumatology department of the Communal Non-Commercial Enterprise of the Lviv Regional Council "Lviv Regional Clinical Hospital" in 2014-2019. To diagnose CMV and EBV infection by enzyme-linked immunosorbent assay, antibodies of IgM and IgG to viruses were detected in blood serum, and viruses were detected by polymerase chain reaction. According to the results of virus detection, formed groups of the patients, namely: patients with active CMV infection, active EBV, active CMV and EBV, without active CMV and EBV. All patients with SLE included in the study were subsequently stratified by age according to the classification of the World Health Organization (2015), according to which the following age limits were determined: young age, middle-aged, elderly, senile. Statistical analysis was performed on a personal computer in MS Excel and Statistica 6.0 using descriptive statistics. The frequency of cases of active CMV and EBV infection was calculated mathematically by the binomial coefficient of I. Newton. Research results and their discussion. We found in the vast majority of patients with SLE (117 patients, 97.50%) increase in the titer of specific antibodies to CMV. Only in 3 patients (2.50%) the titer of antibodies to this virus was within normal limits. Analyzing the frequency of EBV infection in patients with SLE, we recorded an increase in the titer of specific antibodies to the virus in 119 patients (99.17%). Among the examined patients with SLE in all (100.00%) found an increase in the titer of antibodies to CMV and / or EBV, of which 97.50% - infected with CMV and 97.17% - infected with EBV. The active phase of CMV and / or EBV infection was detected in 54.17%, of which 23.33% - active CMV infection, 17.50% - active EBV infection and 12.50% - a combination of active CMV and EBV infection simultaneously, which indicates a high frequency of CMV and EBV infection in patients with SLE and reflects the urgency of the problem of diagnosing herpesvirus infection in them. We found that activeCMV, EBV infections and their combinations are present only in women (64 patients, which is 60.96% of the total number of women with SLE), of which 28 patients (26.67%) there was only active CMV infection, in 21 patients (20.00%) - only active EBV infection and in 15 patients (14.29%) – combination of active CMV and EBV infection. 41 women (39.05%) and all (100.00%) men were not found to have active CMV and EBV infection, which indicates that men at the time of the survey were significantly more likely to have this infection in the integration phase. The most frequently active EBV infection was detected in patients with SLE of young age (17 cases, 24.64%), and in middle-aged patients 3 cases (6.52%) were recorded, which indicates a significant (p <0.05) difference in the frequency of cases of active EBV infection in patients of both groups. Only 1 case (20.00%) of active EBV infection was detected in elderly patients. Conclusions. All patients with systemic lupus erythematosus are infected - 97.50% with cytomegalovirus and 97.17% with Epstein-Barr virus infection, that was confirmed by the increased titer of antibodies to them. Among the mentioned patients 53.33% of them had the active phase of infection (23.33% - cytomegalovirus infection in the replication phase, 17.50% - the Epstein- Barr virus infection in the replication phase and 12.50% - their combination). The prevalence of active viral infection in patients with systemic lupus erythematosus depends on gender (active cytomegalovirus, active Epstein-Barr virus infection and their combination are significantly more common in women) and age - they are probably more common in young patients.

Frequency of cytomegalovirus and Epstein–Barr infections, and their combination in patients with systemic lupus erythematosus depending on the duration of the disease and the degree of its activity

Annotation. Systemic lupus erythematosus (SLE) is a chronic disease characterized by a variety of clinical manifestations with phases of exacerbation and remission, which often complicate the clinician in determining treatment tactics or possible impact on its course of comorbid lesions, including those caused by viral infection. It is known that more than 90.0 % of patients with SLE are infected with cytomegalovirus (CMV) and about 99.5 % with Epstein–Barr virus (EBV). Because CMV and EBV can persist in the human body, more and more attention is being paid to elucidating the effects of active viral infection on the course and activity of SLE, the main pathogenetic mechanism of which is the autoimmune process. The aim of the study was to establish the frequency of cytomegalovirus and Epstein–Barr infections, and their combination in patients with systemic lupus erythematosus depending on the duration of the disease and the degree of its activity. The study involved 120 patients with SLE aged 18 to 69 years, who were in the rheumatology department of the Communal Non-Profit Enterprise of the Lviv Regional Council “Lviv Regional Clinical Hospital” in 2014–2019. To diagnose CMV and EBV infection, antibodies of immunoglobulins M and immunoglobulins G to viruses were detected in blood serum, as well as viruses were detected with polymerase chain reaction and groups of patients were formed: patients with active CMV infection; active EBV; active CMV and EBV; without active CMV and EBV. To achieve the goal, the study was conducted in two stages: determining the frequency of CMV and EBV infection and their combination depending on the duration of SLE (first) and the degree of its activity (second). Statistical analysis was performed in MS Excel using descriptive statistics, Student's t-test to compare samples with normal distribution, χ2-test, z-test to compare two particles; statistically significant relationship was considered when p<0.05. According to our results, active CMV infection is more common in patients with systemic lupus erythematosus duration less than 1 year, active EBV infection – in patients with disease duration greater than 10 years, a combination of active CMV and EBV infection – in patients with systemic lupus erythematosus duration less than 1 year. The increase in the degree of activity of systemic lupus erythematosus is accompanied by a significant (p<0.05) increase in the incidence of active CMV and EBV infection, as well as an increase in mean IgM and IgG antibody titers to these viruses in patients with increasing disease activity.

Antibody-Mediated Immune Subset Depletion Modulates the Immune Response in a Rabbit (Oryctolagus cuniculus) Model of Epstein-Barr Virus Infection.

Epstein-Barr Virus (EBV) is a γ-herpesvirus which infects over 90% of the adult human population. Most notably, this virus causes infectious mononucleosis but it is also associated with cancers such as Hodgkin and Burkitt lymphoma. EBV is a species-specific virus and has been studied in many animal models, including nonhuman primates, guinea pigs, humanized mice, and tree shrews. However, none of these animal models are considered the "gold standard" for EBV research. Recently, rabbits have emerged as a viable alternative model, as they are susceptible to EBV infection. In addition, the EBV infection progresses after immune suppression with cyclosporine A (CsA), modeling the reactivation of EBV after latency. We sought to refine this model for acute or active EBV infections by performing antibody-mediated depletion of certain immune subsets in rabbits. Fourteen 16 to 20-wk old, NZW rabbits were intravenously inoculated with EBV and concurrently treated with either anti-CD4 T-cell antibody, anti-pan-T-cell antibody (anti CD45), CSA, or, as a control, anti-HPV antibody. Rabbits that received the depleting antibodies were treated with CsA 3 times at a dose of 15 mg/kg SC once per day for 4 d starting at the time of EBV inoculation then the dose was increased to 20 mg/kg SC twice weekly for 2 wk. Weights, temperatures, and clinical signs were monitored, and rabbits were anesthetized once weekly for blood collection. When compared with the control group, anti-CD4-treated rabbits had fewer clinical signs and displayed higher levels of viral DNA via qPCR in splenocytes; however, flow cytometry results showed only a partial depletion of CD4 T-cells. Treatment with anti-pan-T-cell antibody did not result in noticeable T-cell depletion. These data suggest the EBV-infected rabbit is a promising model for testing antiviral medications and prophylactic vaccines for EBV.

Epstein-Barr virus (<i>Herpesviridae: Gammaherpesvirinae: Lymphocryptovirus: Human gammaherpesvirus 4</i>): replication strategies

The Epstein-Barr virus (EBV), one of the most common in the human population, is capable of lifelong persistence in resting memory B-cells, in T-cells in case of type 2 EBV, and in some undifferentiated epithelial cells. In most people, EBV persistence is not accompanied by significant symptoms, but frequent virus activations are associated with the increased risks of severe diseases, such as chronic active Epstein-Barr virus infection, hemophagocytic lymphohistiocytosis, multiple sclerosis, systemic lupus erythematosus, gastric and nasopharyngeal carcinomas, and a variety of T- and B-cell lymphomas. Therefore, the molecular viral and host cell processes during asymptomatic or low-symptom EBV persistence are of great interest. This review describes the behavior of the viral DNA in an infected cell and the forms of its existence (linear, circular episome, chromosomally integrated forms), as well as methods of EBV genome copying. Two closely related cycles of viral reproduction are considered. Lytic activation is unfavorable for the survival of a particular viral genome in the cell, and may be a result of differentiation of a latently infected cell, or the arrival of stress signals due to adverse extracellular conditions. The EBV has a large number of adaptive mechanisms for limiting lytic reactivation and reducing hostility of host immune cells. Understanding the molecular aspects of EBV persistence will help in the future develop more effective targeted drugs for the treatment of both viral infection and associated diseases.

Chronic Active Epstein-Barr Virus Infection in a Patient with Enteritis, Myositis, and Pericardial Effusion: A Case Report

Introduction: Chronic active Epstein-Barr virus infection (CAEBV) is a rare, life-threatening disease entity involving multiple organs and systems. The disease could be presented in a single organ with atypical symptoms, be misdiagnosed at the early phase, and resolved without aggressive treatment. Previous studies reported favorable outcomes with early diagnosis and treatment, while in other cases, disease progression and presenting in other organs could result in a very poor outcome. Clinicians from any department should be aware of the CAEBV entity when an unresolved systemic inflammation presents. Case Presentation: A 61-year-old female presented to the cardiology department due to edema and fatigue with the progression of symptoms, including fever, muscle weakness, and shortness of breath. MR images showed muscular edema, and ultrasound analysis revealed massive pericardial effusion. Results of laboratory tests indicated hematological involvement, liver damage, and positivity for EBV DNA, and EBER in muscle and intestine by staining. She was first misdiagnosed with ulcerative colitis, but the disease evolved to the muscles and cardiovascular system later on. Altogether, the CAEBV infection with enteritis, myositis, and pericardial effusion was confirmed in the case. Although diagnosed, the treatment of the elderly population remains rather challenging in severe cases. The patient died after disease onset for less than three years. Conclusions: Elderly patients with CAEBV infection tend to have a poor prognosis. Early diagnosis would facilitate early treatment and the utilization of available options, which might reduce the disease mortality rate eventually. Individualized therapy and promising approaches should be further developed for CAEBV infection.

Rapid identification and characterization of infected cells in blood during chronic active Epstein-Barr virus infection.

Epstein-Barr virus (EBV) preferentially infects epithelial cells and B lymphocytes and sometimes T and NK lymphocytes. Persistence of EBV-infected cells results in severe lymphoproliferative disorders (LPDs). Diagnosis of EBV-driven T or NK cell LPD and chronic active EBV diseases (CAEBV) is difficult, often requiring biopsies. Herein, we report a flow-FISH cytometry assay that detects cells expressing EBV-encoded small RNAs (EBERs), allowing rapid identification of EBV-infected cells among PBMCs. EBV-infected B, T, and/or NK cells were detectable in various LPD conditions. Diagnosis of CAEBV in 22 patients of Caucasian and African origins was established. All exhibited circulating EBV-infected T and/or NK cells, highlighting that CAEBV is not restricted to native American and Asian populations. Proportions of EBV-infected cells correlated with blood EBV loads. We showed that EBV-infected T cells had an effector memory activated phenotype, whereas EBV-infected B cells expressed plasma cell differentiation markers. Thus, this method achieves accurate and unambiguous diagnoses of different forms of EBV-driven LPD and represents a powerful tool to study their pathophysiological mechanisms.

Chronic active Epstein-Barr virus infection involving gastrointestinal tract mimicking inflammatory bowel disease

BackgroundChronic active Epstein-Barr virus infection (CAEBV) is a rare disease, which is difficult to be differentiated from inflammatory bowel disease (IBD). To cause the attention, we present twelve cases of CAEBV in immunocompetent patients with gastrointestinal tract involvement.MethodsTwelve patients who fulfilled the diagnostic criteria of CAEBV were enrolled in this retrospective study. The control group was consisted of twenty-four IBD patients with EBV-DNA value increased in peripheral blood. The clinicopathologic and endoscopic characteristics were reviewed and analyzed.ResultsThe major clinical presentations of CAEBV patients were intermittent fever (100%), hepatomegaly/splenomegaly (58%), lymphadenopathy (50%), diarrhea (50%) and hematochezia (50%). Compared with IBD patients, the incidence of intermittent fever and increased level of ferritin were significantly higher among CAEBV patients. The median values for EBV detected in peripheral blood were significantly higher in CAEBV group (1.42*10^6 copies/μg) than in IBD group (3.2*10^3 copies/μg, p<0.05). The main endoscopic findings of CAEBV included multifocal or isolated, irregular, multiform ulcers and diffuse inflammation, lacking of typical cobblestone appearance. Ten patients died within 5 years of disease onset. The average survival time is 21 months.ConclusionsSymptoms such as intermittent fever, increased level of ferritin and atypical endoscopic findings could be a sign for CAEBV. Early detections of EBV-DNA in serum and EBV-encoded small nuclear RNA (EBER) by in situ hybridization in intestinal tissue are essential for differential diagnosis between CAEBV and IBD.

Evaluation of Epstein-Barr Virus Indirect Immunofluorescence Assay Results

Aim: Epstein-Barr Virus (EBV) causes Infectious Mononucleosis (IM) and chronic active EBV infections, malignant diseases such as Burkitt’s lymphoma, nasopharyngeal carcinoma, and posttransplant lymphoproliferative disease. More than 90% of children are infected with EBV until the age of six. The aim of this study was to diagnose EBV infection using Indirect Immunofluorescence Assay (IIFA) method and to evaluate these patients clinically. Methods: The tests were studied by using EBV Indirect Immunofluorescence Assay (IIFA) method. A total of 247 patients, 186 adults, and 61 children, were included in the study. Results: Five (2.7%) of the adults were EBV-Capsid Antigen (EBV-CA) IgM positive, 175 (94.1%) were IgG positive and 6 (3.2%) were seronegative. 10 (16.4%) of the child patients were IgM positive, which is considered as an acute IM infection marker, whereas the child patients had a significantly higher IgM rate than adults (p &lt;0.001). 39 (63.9%) of the child patients were IgG positive and 12 (19.7%) of them were seronegative. The rate of IgG positivity in children was significantly lower than in adults (p &lt;0.001). There was no significant difference between the genders in terms of IgM and IgG positivity rates in both adults and children (p &gt;0.05 for each). Conclusion: These results suggest that the presence of an acute EBV infection should be considered when the patient has viral diseases with similar clinical picture especially for children.

Nationwide survey of systemic chronic active EBV infection in Japan in accordance with the new WHO classification

AbstractSystemic chronic active Epstein-Barr virus infection (sCAEBV) was defined as a T- or NK-cell neoplasm in the 2017 World Health Organization (WHO) classification. To clarify the clinical features of sCAEBV under this classification and review the effects of chemotherapy, we performed a nationwide survey in Japan from 2016 through 2018 of patients with sCAEBV newly diagnosed from January 2003 through March 2016. One hundred cases were evaluated. The patients were aged 1 to 78 years (median, 21) and included 53 males and 47 females. Spontaneous regression was not observed in patients with active disease. In the childhood-onset group (age, <9 years), 78% of the patients were male. In contrast, 85% of the patients in the elderly-onset group (age, >45 years) were female. The prognosis of the childhood-onset group was better than those of the adolescent/adult- and elderly-onset groups. The main chemotherapies used were a combination of cyclosporine A, steroids, and etoposide (cooling therapy) in 52 cases and cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP) in 45 cases. The rate of complete response (CR), defined as complete resolution of disease activity, was 17% for cooling therapy and 13% for CHOP. Virological CR was not observed. The 3-year overall survival rates in patients treated with chemotherapy only (n = 20), chemotherapy followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT; n = 47), and allo-HSCT only (n = 12) were 0%, 65%, and 82%, respectively. Distinct characteristics were observed between childhood- and elderly-onset sCAEBV, and they appeared to be different disorders. Chemotherapy is currently insufficient to resolve disease activity and eradicate infected cells. The development of an effective treatment is urgently needed.

Clinical characteristics of primary and reactivated Epstein-Barr virus infection in children.

Epstein-Barr virus (EBV) infection occurs commonly in children and presents as a primary or reactivated infection, which are difficult for clinicians to distinguish. This study investigated the clinical characteristics of the two types of infections. Children with detectable plasma EBV-DNA were retrospectively enrolled and divided into primary and reactivated infection group by EBV-specific antibody. We analyzed the patients' characteristics, clinical manifestations, complications, inflammatory biomarkers, and viral load. A total of 9.3% of children with reactivation were immunocompromised over the long-term. The primary infection mostly appeared as infectious mononucleosis (99.8%), while reactivation occurred as an infectious mononucleosis-like disease (65.0%), hemophagocytic syndrome (22.6%), chronic active EBV infection (5.3%) and lymphoma (3.5%). The incidence of fevers, cervical lymphoditis, periorbital edema, pharyngotonsillitis, hepatomegaly and splenomegaly in primary infection were 93.3%, 93.0%, 51.5%, 66.0%, 76.2% and 63.9%, respectively; the incidence of those symptoms in reactivation was 84.0%, 46.9%, 15.4%, 18.5%, 18.5%, and 43.3%, respectively. The incidence of digestive, respiratory, cardiovascular, neurological, hematological, genitourinary complications and multiple serous effusion in primary infection was 68.8%, 18.1%, 8.0%, 0.8%, 2.9%, 0.0% and 2.3%; whereas the incidence of these complications in reactivation was 56.2%, 22.5%, 14.1%, 8.0%, 38.9%, 0.3% and 19.0%. Patients with reactivation were more prone to multi-systemic damage. B-cells were lower, and CD8+ T-cells were higher in primary infection. Viral load was correlated with the level of different cytokines in primary and reactivated infection. EBV primary infection often presents as infectious mononucleosis. The reactivated infection affects more immunocompromised subjects with diverse and complex manifestations. Various complications are more commonly associated with reactivation as a result of different inflammatory responses to different types of infection.

The aim of the measurement of Epstein-Barr virus DNA in hydroa vacciniforme and hypersensitivity to mosquito bites.

Epstein-Barr virus (EBV) DNA load in the blood increases in posttransplant lymphoproliferative disorders and chronic active EBV infection. In this report, we analyzed the EBV DNA load in the peripheral blood mononuclear cells (PBMCs) and plasma of patients with hydroa vacciniforme (HV) and/or hypersensitivity to mosquito bites (HMB) to understand the clinical significance of EBV DNA load. All 30 patients showed high DNA loads in the PBMCs over the cut-off level. Of 16 plasma samples, extremely high in two samples obtained from patients with hemophagocytic lymphohistiocytosis (HLH). The amount of cell-free DNA in plasma was correlated to the serum levels of lactate dehydrogenase and inversely correlated to platelet counts. These results indicate that the EBV DNA load in PBMCs can provide one of the diagnostic indicators for HV and HMB and marked elevation of cell-free EBV DNA in plasma might be related to cytolysis such as that observed in HLH.