Absence Of Epstein-Barr Virus Research Articles

Epstein-Barr Virus and multiple sclerosis in a Spanish cohort: A two-years longitudinal study.

Objectives1. To analyze the prevalence and levels of anti-EBNA-1 and anti-VCA IgG antibodies of Epstein-Barr virus (EBV) in a Spanish cohort of multiple sclerosis (MS) patients and their interactions with other environmental and genetic risk factors. 2. To analyze the association of the evolution of these antibodies with the clinical response to different disease modifying therapies (DMTs) after two-years of follow-up. 3. To assess their possible correlation with the class II HLA alleles as well as with several SNPs identified in GWAS related to disease susceptibility.Materials and methodsWe recruited 325 MS patients without DMT (serum samples were collected 1-3 months before starting a therapy) and 295 healthy controls (HC). For each patient we also collected serum samples 6, 12, 18 and 24 months after starting the DMT. EBNA-1 and VCA IgG titers were analyzed by ELISA; 25(OH)D levels were analyzed by immunoassay; HLA DRB1*15:01 allelic variant was analyzed by Taqman technology.Results1. 97.8% (318/325) vs. 87.1% (257/295) positives for EBNA-1 in MS patients and HC, respectively (p<0.0001; O.R. = 6.7); 99.7% (324/325) vs. 94.6% (279/295) for VCA in MS patients and HC, respectively (p=0.0001; O.R. = 18.6). All MS patients were positive for EBNA-1 and/or VCA IgG antibodies vs. 280/295 (94.9%) HC (p<0.0001). IgG titers were also significantly higher in MS patients than in HC. 2. We did not find any statistical correlation in the variation of the EBNA-1 and VCA IgG titers between baseline and 24 month visits with the number of relapses, progression, clinical response, NEDA-3 condition or therapeutic failure. 3. When we compared different epidemiological and clinical variables between those with genetic factors associated with lower EBNA-1 IgG titers and all other MS patients, we found MS started 3.5 years later among the first.ConclusionsThese results confirm that MS occurs rarely in absence of EBV. An intriguing association between genetic burden and lower EBNA-1 IgG titers was associated with an earlier age of disease onset. Similar studies with B-cell–targeted therapies should be performed.

Dimorphism in the TCRγ-chain repertoire defines 2 types of human immunity to Epstein-Barr virus

Humans form 2 groups based on their innate immunity to Epstein-Barr virus (EBV). Group 1 makes a strong natural killer (NK)-cell and γδ T-cell response, whereas group 2 makes a strong NK-cell response, but a weak γδ T-cell response. To investigate the underlying basis for this difference in γδ T-cell immunity to EBV, we used next-generation sequencing to compare the γδ T-cell receptor (TCR) repertoires of groups 1 and 2. In the absence of EBV, group 1 TCRγ chains are enriched for complementarity determining region 3 (CDR3s) containing JγP, whereas group 2 TCRγ chains are enriched for CDR3s containing Jγ2. In group 1 donors, EBV activates many γδ T cells expressing Vγ9JγP, inducing proliferation that produces a large population of activated effector cells. The TCRs using Vγ9JγP are closely related to the TCRs of γδ T cells that respond to phosphoantigens. In group 2 donors, EBV activates a small subpopulation of γδ T cells, most expressing Vγ9JγP. In conclusion, we find that differences in the TCRγ-chain repertoire underlie the differential response of group 1 and group 2 to EBV.

The prognosis of chronic erosive gastritis associated with Helicobacter pylori and Epstein-Barr

Аim. To study the severity of chronic erosive gastritis associated with Helicobacter pylori and Epstein-Barr virus depending on the number of copies of Epstein-Barr virus in biopsies of gastric mucosa. Materials and methods. The severity of chronic erosive gastritis associated with Helicobacter pylori in the presence and absence of Epstein-Barr virus in the gastric mucosa was compared in 65 patients, divided into 4 groups depending on the detection of Epstein-Barr virus in the gastric mucosa. Result. the presence of Epstein-Barr virus in the gastric mucosa aggravates the course of chronic erosive gastritis associated with Helicobacter pylori.

Non-Destructive Post-Transplant Lymphoproliferative Disorder Shows Distinct Clinical Features with HHV-6 Reactivation in a Subset

Background: Post-transplant lymphoproliferative disorders (PTLD) are a diverse group of lymphoid neoplasms arising after solid organ (SOT) or hematopoietic stem cell transplant (HSCT). They may resemble lymphomas in immunocompetent patients (monomorphic); present as destructive, heterogeneous lesions (polymorphic); or be non-destructive but mass-forming (non-destructive, previously known as 'early lesions'). PTLDs are often associated with Epstein-Barr virus (EBV) infection or reactivation, which likely functions as a driver in these cases. Non-destructive lesions may be challenging to diagnose in the absence of EBV. However, a significant fraction of PTLDs are EBV(-), including a subset of non-destructive lesions. The pathogenesis of these remains to be fully elucidated. Human herpesvirus 6 (HHV-6) reactivation is common during transplantation, occurring in up to 70% of HSCT and 80% of SOT patients. While it has been hypothesized that HHV-6 may play a role in PTLD, particularly in EBV(-) cases, the role of HHV-6 in the pathogenesis of PTLD remains underexplored. Methods: Following IRB approval, pathology archives at Weill Cornell/New York Presbyterian Hospital were searched to identify patients with a tissue diagnosis of PTLD and correlated with clinical information. A tissue microarray was constructed from blocks with sufficient tissue. Standard immunohistochemistry (IHC) and in situ hybridization for EBV-encoded RNA (EBER) were performed, and IHC conditions were optimized for antibodies against HHV-6 gp60/110 (Millipore Sigma, MAB853) and p41 (Santa Cruz Biotechnologies, 9A5D12). Statistical comparisons utilized Student's t-test and Fisher's exact test. Results: We identified 79 patients from 6/23/94-6/6/19 with a tissue diagnosis of PTLD. Morphologic subtype could be determined in 74, including 8 patients with non-destructive lesions. Compared to monomorphic or polymorphic, patients with non-destructive PTLDs were significantly younger (median age 9, p&lt;0.01) and more likely status post heart transplant (5/8 cases, p&lt;0.001, Table 1). By contrast, polymorphic PTLDs occurred relatively more frequently in HSCT patients (25/33 cases, p&lt;0.01). All morphologic subtypes were frequently EBV+ (63-88%) but EBV positivity was greater in polymorphic than monomorphic lesions (p&lt;0.05). While tissue testing failed on blocks &gt; 20 years old, 13 PTLD cases from 12 patients had sufficient evaluable material for HHV-6 testing (Table 2). 9/12 patients had a SOT (8 renal, 1 heart) and 3 had HSCT. HHV-6 was identified in 5/13 lesions (38%), all of which were in SOT patients (Figure 1). EBER was detected in 8/13 lesions (62%). When stratified by PTLD subtype, 3/3 non-destructive PTLDs were HHV-6+ (100%). Two were EBER(-) and the remaining case only showed scattered EBER+ cells. Unfortunately, material was not available for testing the EBER+ non-destructive lesions. 2/9 of the monomorphic lesions tested (22%) were positive for both HHV-6 and EBER. While the series is limited, the association between HHV-6 and non-destructive lesions, compared to other types of PTLD, appears significant (p&lt;0.05). EBV did not appear to be associated with HHV-6 status (p = 0.29). Conclusions: Non-destructive, monomorphic and polymorphic PTLDs show significant differences in timing, patient demographics and transplant type, suggesting that they may differ in associated risk factors affecting their pathogenesis. Confirming prior studies, we find non-destructive lesions tend to occur in younger patients, often following SOT as compared to HSCT and with a median time from transplantation of ~50 months. In addition, the presence of HHV-6 in the absence of (or with limited) EBV in the 3 non-destructive lesions tested raises the possibility that HHV-6 may play a role in the pathogenesis of this PTLD subtype that has yet to be defined. These findings are intriguing given the frequent reactivation of HHV-6 seen in transplant populations. HHV-6 has been variably associated with post-transplant complications including pneumonitis, delayed hematopoietic engraftment and graft versus host disease and thus received relatively more attention in the HSCT population. A causative role for HHV-6 in PTLD remains speculative, but these findings raise the possibility that clinicians treating the SOT population should also consider aggressively identifying and treating HHV-6 reactivation. Disclosures Rutherford: AstraZeneca: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; Heron: Consultancy, Honoraria; Janssen Scientific Affairs: Consultancy, Honoraria; Juno Therapeutics Inc: Consultancy, Honoraria; Seattle Genetics: Consultancy, Honoraria; Verastem: Consultancy, Honoraria; Karyopharm: Honoraria, Membership on an entity's Board of Directors or advisory committees.

Traumatic ulcerative granuloma with stromal eosinophilia\xa0\u2014\xa0clinical case report, literature review, and differential diagnosis

BackgroundTraumatic ulcerative granuloma with stromal eosinophilia (TUGSE) is a rare self-limiting condition of the oral mucosa. The lesion manifests as an isolated ulcer that can be either asymptomatic or associated with mild to severe pain, and in most cases, it affects the tongue. TUGSE lesions may mimic malignancy such as squamous cell carcinoma, CD30 positive lymphoproliferative disorder, or infectious diseases such as primary syphilis, tuberculosis, or Epstein-Barr virus mucocutaneous ulcer. Histologically dominating cells are lymphocytes, histiocytes, and eosinophils.Case presentationWe describe a TUGSE case of a patient with a solitary ulcer on the lower left retromolar buccal plane. Upon presentation, the patient reported a swelling on the buccal mucosa of the left lower jaw since 1 year with rapid growth over the last days and mild pain while chewing. The diameter of the intraoral lesion on the lower left retromolar buccal plane was approximately 4 × 3 cm; the lesion presented as indurated base with a central superficial ulceration of 2 × 1 cm, indicative for a malignant process. Histologically, the ulceration showed an expanding, infiltrative, and vaguely granulomatous morphology, involving the superficial mucosa and the fatty tissue, and extended between the deep striated muscle fibers. The lesion was rich in lymphocytes, histiocytes, and eosionophils intermingled with activated T-blasts without phenotypic abnormalities. TUGSE was then diagnosed based on the phenotype (especially the lacking expression of CD30, the retained T-cell phenotype, and the absence of Epstein-Barr virus), the clinical presentation, and the morphology. Twenty-six months after diagnosis, no recurrence of the ulceration was seen.ConclusionsAs TUGSE may mimic malignancy or infectious diseases, biopsy is mandatory and should be combined with thorough clinical examination. A screening for infectious diseases (mainly syphilis, Epstein-Barr virus, and HIV infections) must be performed routinely. In most cases, the lesions resolve spontaneously, obviating the need of further actions other than clinical follow-up. The pathogenesis of TUGSE lesions is still under debate, although local traumatic events and a locotypic immune response have been suggested to be major contributing factors.

DNA Damage Signaling Is Induced in the Absence of Epstein-Barr Virus (EBV) Lytic DNA Replication and in Response to Expression of ZEBRA.

Epstein Barr virus (EBV), like other oncogenic viruses, modulates the activity of cellular DNA damage responses (DDR) during its life cycle. Our aim was to characterize the role of early lytic proteins and viral lytic DNA replication in activation of DNA damage signaling during the EBV lytic cycle. Our data challenge the prevalent hypothesis that activation of DDR pathways during the EBV lytic cycle occurs solely in response to large amounts of exogenous double stranded DNA products generated during lytic viral DNA replication. In immunofluorescence or immunoblot assays, DDR activation markers, specifically phosphorylated ATM (pATM), H2AX (γH2AX), or 53BP1 (p53BP1), were induced in the presence or absence of viral DNA amplification or replication compartments during the EBV lytic cycle. In assays with an ATM inhibitor and DNA damaging reagents in Burkitt lymphoma cell lines, γH2AX induction was necessary for optimal expression of early EBV genes, but not sufficient for lytic reactivation. Studies in lytically reactivated EBV-positive cells in which early EBV proteins, BGLF4, BGLF5, or BALF2, were not expressed showed that these proteins were not necessary for DDR activation during the EBV lytic cycle. Expression of ZEBRA, a viral protein that is necessary for EBV entry into the lytic phase, induced pATM foci and γH2AX independent of other EBV gene products. ZEBRA mutants deficient in DNA binding, Z(R183E) and Z(S186E), did not induce foci of pATM. ZEBRA co-localized with HP1β, a heterochromatin associated protein involved in DNA damage signaling. We propose a model of DDR activation during the EBV lytic cycle in which ZEBRA induces ATM kinase phosphorylation, in a DNA binding dependent manner, to modulate gene expression. ATM and H2AX phosphorylation induced prior to EBV replication may be critical for creating a microenvironment of viral and cellular gene expression that enables lytic cycle progression.

Simultaneous Occurrence of Angioimmunoblastic T-cell Lymphoma and Plasma Cell Leukemia

A 73-yr-old man was evaluated for fever and multiple palpable nodules in his neck and inguinal areas. Biopsy of a cervical lymph node demonstrated polymorphic infiltrates including T-cells with abundant clear cytoplasm (clear cells), follicular dendritic cells, and plasma cells in the background of prominent arborizing high endothelial venules. The involved lymph node had typical histological and immunohistochemical properties characteristic of angioimmunoblastic T-cell lymphoma (AITL), except for the absence of CD10 expression. Lymphoplasmacytic lymphoma was excluded. A staging computed tomographic scan showed generalized lymphadenopathy in his chest, abdomen, and pelvis, as well as hepatosplenomegaly. Laboratory findings demonstrated the presence of leukocytosis (30.5×109/L). A peripheral blood (PB) smear revealed rouleaux formations and numerous circulating plasma cells, with plasmacytoid lymphocytes accounting for 25% of all nucleated cells. Protein electrophoresis and immunofixation studies of serum indicated IgMκ monoclonal gammopathy (IgM level, 53.8 g/L; normal range, 0.4-2.3 g/L). A bone marrow (BM) biopsy revealed 80% hypercellularity with focal clusters of CD3+ malignant T-cells and diffuse CD138+ clonal plasma cells at proportions of 10% and 30%, respectively (Fig. 1). The CD3+ T-cells were medium-sized lymphoid cells with moderate amounts of clear cytoplasm and slightly indented nuclei with coarse and condensed chromatin. In concordance with the monoclonal gammopathy found in the serum, immunohistochemical analysis of mixed plasma cells revealed a pattern restricted to κ light chain reactivity. Rare scattered cells were found positive for Epstein-Barr virus (EBV) by using in situ hybridization. Analysis for the expression of CD20 indicated relatively few mature B-cells. Fig. 1 Histologic findings of the bone marrow biopsy. (A) Bone marrow biopsy showed nodular lymphocytic infiltrates with malignant cells (Hematoxylin and Eosin, ×400). (B) The malignant cells were positive for CD3 (×400). (C, D) Plasma cells ... Flow cytometry analysis of circulating lymphocytes revealed a B-cell population expressing CD19, but the cells lost CD20 without surface Ig expression (25.12% of total events). Instead, these cells intensely expressed CD38 with a fraction of the cells also expressing CD138. Almost all CD38+ cells exhibited cytoplasmic κ restriction. In addition to the clonal plasma cells, T-cells that co-expressed CD3 and CD4 were also found to be positive for the known T-cell antigens CD2 and CD5 but negative for CD7. Consistent with the immunohistological findings from primary lymph node, T-cells were found to be negative for CD10. T-cells with immunophenotypic variation were considered AITL cells and comprised up to 0.25% of the total gated events. Representative scatter plots of the immunophenotypic findings obtained from the flow cytometry analysis are shown in Fig. 2. Fig. 2 Flow cytometry analysis of circulating lymphocytes. (A) and (B) CD45 and side scatter (SSC) gating: T-cells were positive for CD3, and B-cells were positive for CD19. (C) and (D) The CD19+ B-cells expressed high density of CD38 and cytoplasmic monoclonal ... PCR analysis of the T-cell receptor gamma chain gene (TCRγ) demonstrated a clonal rearrangement that confirms T-cell clonality. Molecular analyses for the TCRγ rearrangement was performed through PCR by using a mixture of consensus V and J region primers. The PCR products were analyzed by nonradioactive single-strand confirmation polymorphism. Karyotypic analysis of the BM cell culture using conventional G-banding showed 46,XY in all metaphase cells analyzed (n=20). Consequently, the patient was diagnosed as having BM involvement of AITL and plasma cell leukemia (IgMκ) simultaneously. Following initial diagnosis, the patient received the first cycle of chemotherapy consisting of vincristine, doxorubicin, cyclophosphamide, and prednisolone. The patient also suffered from respiratory failure combined with septic shock and was managed at the intensive care unit. Following discharge, the patient was followed up to monitor the disease in an outpatient clinic, without further chemotherapy. AITL, one subtype of peripheral T-cell lymphoma, is frequently associated with polyclonal B-cell or plasma cell proliferation [1]. The B-cell expansion observed in AITL has been proposed to be related to the functions of neoplastic follicular helper T-cells. However, proliferation of clonal B-cells or plasma cells has rarely been reported [2, 3, 4]. We hypothesized that the clonal plasma cell population is unrelated to the patient's AITL status. In over 20% of clonal plasma cells in PB and BM, the presence of M-protein in serum and absence of EBV could support this possibility. Factors contradicting plasma cell neoplasm include the followings: negative organ impairment including hypercalcemia; renal insufficiency; anemia; bone lesions; and IgM paraprotein (non-IgG or IgA) that could be associated with a clone of lymphoplasmacytic cells [5]. However, there was no evidence of CD20+ neoplastic B-cells in BM. In conclusion, the current case emphasizes that AITL could present with marked proliferation of clonal plasma cells, thereby potentially masking underlying T-cell lymphoma. Being aware of this possible effect is clinically important when determining treatment strategies and prognostic stratification, especially in the patients with a clonal B-cell population.

Epstein-barr virus-negative post-transplant lymphoproliferative diseases: three distinct cases from a single center.

Three cases of Epstein-Barr virus (EBV)-negative post-transplant lymphoproliferative disease that occurred 6 to 8 years after renal transplantation are reported. The patients respectively had gastric mucosa-associated lymphoid tissue lymphoma, gastric diffuse large B-cell lymphoma, and atypical Burkitt lymphoma. Absence of EBV in the tissue samples was demonstrated by both in situ hybridization for EBV early RNA and polymerase chain reaction for EBV DNA. Patients were treated with reduction in immunosuppression and combined chemotherapy plus an anti-CD20 monoclonal antibody, rituximab. Despite the reduction in immunosuppression, patients had stable renal functions without loss of graft functions. The patient with atypical Burkitt lymphoma had an abnormal karyotype, did not respond to treatment completely, and died due to disease progression. The other patients are still alive and in remission 5 and 3 years after diagnosis, respectively. EBV-negative post-transplant lymphoproliferative diseases are usually late-onset and are reported to have poor prognosis. Thus, reduction in immunosuppression is usually not sufficient for treatment and more aggressive approaches like rituximab with combined chemotherapy are required.

Atypical interfollicular hyperplasia of tonsils resembling mucosa-associated lymphoid tissue lymphoma: a clinicopathological, immunohistochemical study and epstein-barr virus findings in 12 cases.

This study attempted to clarify the clinicopathological and immunohistochemical findings and presence or absence of Epstein-Barr virus (EBV) in tonsillar atypical interfollicular hyperplasia (AIFH). A total of 597 consecutive specimens from tonsillectomies performed in Dokkyo University School of Medicine between 1999 and July 2013 were reexamined. Using formalin-fixed, paraffin-embedded sections, histological and immunohistochemical analyses, and in situ hybridization (ISH) were performed. AIFH was identified in the tonsils in 12 (2.0%) cases. These included 7 males and 5 females, aged 3 to 19 years (mean, 7). Histologically, there was expansion of the interfollicular areas by polymorphous infiltration resulting in distortion, but not obliteration of the normal tonsillar architecture. In some areas, the lymphoid follicles had hyperplastic germinal centers with ill-defined borders surrounded by sheet-like proliferation of polymorphous infiltrate showing a marginal zone distribution pattern. The infiltrate was composed of small to medium-sized (transformed) lymphocytes and immunoblasts accompanied by numerous plasma cells and plasmacytoid cells, and resembling monocytoid B cells. The numerous immunoblasts were MUM1(+), CD10(-), BCL-6(-). An ISH study demonstrated EBV-encoded small RNA-1 (EBER-1)(+) cells in 9 lesions. Moreover, in 3 lesions, numerous EBER-1(+) cells were present in germinal centers as well as in interfollicular areas. The present study indicated that EBV may cause at least a portion of tonsillar AIFH in children and adolescents. In conclusion, an atypical lymphoid infiltration resulting in distortion of tonsillar architecture with numerous MUM1(+), CD10(-), BCL-6(-) immunoblasts should raise the suspicion of a reactive process.

Screening of Ameloblastoma Cases in Ibadan for HPV and EBV Genes

This study investigated the presence or absence of Epstein-Barr virus and Human papilloma virus genes in ameloblastoma, in order to elucidate the genetic origin of this tumour. Methods and Material: 89 archived formalin-fixed paraffin-embedded specimens of ameloblastoma from the year 2000-2010 were genotyped by polymerase chain reaction for Epstein-Barr virus and Human papilloma virus genes Results: None of the 89 ameloblastoma cases used for this study tested positive for Epstein-Barr virus and Human papilloma virus genes. Conclusions: This study suggested that Epstein-Barr virus and Human papilloma virus genes may likely not be involved in the aetiopathogenesis of ameloblastoma in Ibadan environment. Key Messages: Studies have suggested that HPV might be implicated in the pathogenesis of ameloblastoma. In this study none of the extracted DNA samples from the 89 ameloblastoma cases amplified for HPV primer in the PCR, as this may possibly clear the role of this virus in the etiopathogenesis of ameloblastoma or may indicate need for other primers. Further investigation should be done to clarify EBV role in ameloblastoma.

Epithelioid Cell Granulomatous Response of Waldeyer's Ring among Japanese : A Clinicopathological and Immunohistochemical Study of 16 Cases

The tonsils are uncommonly affected by granulomatous inflammation. This study attempted to clarify the clinicopathological and immunohistochemical findings and presence or absence of Epstein-Barr virus (EBV) in tonsilar granulomatous inflammation. A total of 537 consecutive specimens from tonsillectomies performed at Dokkyo University School of Medicine between 1999 and March 2012 were reexamined. Using formalin-fixed, paraffin-embedded sections, histological, immunohistochemical, and in situ hybridization (ISH) studies were performed. Epithelioid granulomas (EPGs) were identified in the tonsils in 16 (3.0%) cases. There were 8 males and 8 females, aged 4 to 57 years (mean, 23). In 11 patients, EPGs were located in the germinal center (GC), whereas they were located in the interfollicular area as well as GC in the remaining 5 cases. Three types of EPG have been delineated : (i) poorly demarcated small epithelioid cell granulomas (n = 6) ; (ii) well-demarcated non-caseating sarcoid-like granulomas (n = 5) ; and (iii) EPGs within GC showing suppurations at the center (n = 5). An ISH study demonstrated EBV-encoded small RNA (EBER)(+) cells in 4 lesions. The present study demonstrated that the majority of EPGs were located in the GC and tonsilar EPGs showed histological variation.

P52 Activation in Monomorphic B-Cell Posttransplant Lymphoproliferative Disorder/Diffuse Large B-Cell Lymphoma without BAFF-R Expression

Posttransplantation lymphoproliferative disorders (PTLD) are associated with Epstein-Barr virus (EBV) and activate the NF-κB pathway. B-cell activating factor (BAFF) modulates cell growth and survival in non-Hodgkin's lymphomas. However, there are few studies of EBV, BAFF/BAFF-R signaling, and NF-κB1 and NF-κB2 pathway activation in PTLD. Diffuse large B-cell lymphomas (DLBCL) in two different clinical contexts, immunocompetent patients (DLBCL/IC; n = 30) or posttransplantation solid-organ recipients (DLBCL/PTLD; n = 21), were characterized histogenically as germinal center (GC) or non-germinal center (NGC). Expression of BAFF, BAFF-R, and NF-κB proteins p50 and p52 and the presence or absence of EBV were compared in these clinical contexts. Regardless of the GC or NGC pattern of DLBCL, BAFF-R was expressed in 37% of DLBCL/IC but in only 4.8% of DLBCL/PTLD. p52 was expressed in DLBCL/PTLD/NGC (12 of 19 cases) as compared with DLBCL/IC/NGC (0 of 18 cases). This pattern might be related to the presence of EBV and latent membrane protein 1 because p52 expression was observed primarily in EBV-positive DLBCL/PTLD cases expressing latent membrane protein 1. Thus, the activation profile or NGC pattern of DLBCL/PTLD was not associated with BAFF/BAFF-R expression, whereas nuclear p52 related to NF-κB2 pathway activation might be linked to EBV.

Bortezomib induces apoptosis in T lymphoma cells and natural killer lymphoma cells independent of Epstein‐Barr virus infection

Epstein-Barr virus (EBV), which infects not only B cells, but also T cells and natural killer (NK) cells, is associated with multiple lymphoid malignancies. Recently, the proteasome inhibitor bortezomib was reported to induce apoptosis of EBV-transformed B cells. We evaluated the killing effect of this proteasome inhibitor on EBV-associated T lymphoma cells and NK lymphoma cells. First, we found that bortezomib treatment decreased the viability of multiple T and NK cell lines. No significant difference was observed between EBV-positive and EBV-negative cell lines. The decreased viability in response to bortezomib treatment was abrogated by a pan-caspase inhibitor. The induction of apoptosis was confirmed by flow cytometric assessment of annexin V staining. Additionally, cleavage of caspases and polyadenosine diphosphate-ribose polymerase, increased expression of phosphorylated IκB, and decreased expression of inhibitor of apoptotic proteins were detected by immunoblotting in bortezomib-treated cell lines. We found that bortezomib induced lytic infection in EBV-positive T cell lines, although the existence of EBV did not modulate the killing effect of bortezomib. Finally, we administered bortezomib to peripheral blood mononuclear cells from five patients with EBV-associated lymphoproliferative diseases. Bortezomib had a greater killing effect on EBV-infected cells. These results indicate that bortezomib killed T or NK lymphoma cells by inducing apoptosis, regardless of the presence or absence of EBV.

Absence of Epstein-Barr virus in the brain and CSF of patients with multiple sclerosis

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus that becomes latent in B-lymphocytes and has been implicated in the pathogenesis of multiple sclerosis (MS). We searched for latent and active EBV infection in MS brain and CSF. Nested and non-nested real-time PCR were used to detect cell-specific and EBV-specific transcripts in 15 fresh-frozen and 5 formalin-fixed paraffin-embedded MS plaques and in single MS CSF B-lymphocytes and plasma cells. Intrathecal anti-EBV antibody synthesis was measured by ELISA. Immunocytochemistry was used to detect binding of MS CSF and recombinant antibodies (rAbs) generated from clonally expanded plasma cells in MS CSF to EBV-infected cells. No EBV RNA was found in MS CSF B-lymphocytes or plasma cells. In active MS plaques, EBV-encoded RNA (EBER)-1 was the only and rarely detected transcript. The frequency of detected intrathecal anti-EBV antibody synthesis in patients with MS did not differ from that in non-MS inflammatory CNS disease control patients. Anti-EBV antibodies were detected in the CSF of patients with MS, but MS rAbs did not react with EBV. Application of real-time PCR to multiple sclerosis brain and single B-lymphocytes in CSF did not reveal any evidence of active Epstein-Barr virus infection.

Cryptogenic organizing pneumonia after rituximab therapy for presumed post-kidney transplant lymphoproliferative disease

Cryptogenic organizing pneumonia (COP, formerly bronchiolitis obliterans organizing pneumonia) is rare in children. We describe an 11-year-old girl with Epstein-Barr virus (EBV) reactivation/presumed post-transplant lymphoproliferative disease (PTLD) 15 months after undergoing a deceased donor kidney transplantation. Treatment with reduced immunosuppression, ganciclovir, and cytomegalovirus immunoglobulin was complicated by severe graft rejection, prompting therapy with methylprednisolone, anti-thymocyte globulin and four weekly doses of rituximab (total 1500 mg/m(2)). Tacrolimus- and prednisone-based anti-rejection prophylaxis was complemented with low-dose sirolimus. When the lactate dehydrogenase and uric acid levels rose 10 weeks after the first rituximab infusion and bilateral pulmonary nodules were detected by computerized tomography, recurrence of PTLD was suspected. Open lung biopsy of the clinically asymptomatic patient identified the nodules as COP, characterized by abundant CD3(+) T-cells, few B-cells, and the absence of EBV, cytomegalovirus, or adenovirus antigens. With normalization of the peripheral B-cell count, EB viremia reappeared and persisted, despite minimal immunosuppression. Four years later, the patient was diagnosed with classical Hodgkin lymphoma-type PTLD with multiple pulmonary and abdominal nodes. This first report of rituximab-associated, pediatric COP highlights the risk of pulmonary complications after treatment with B-cell depleting agents in solid organ transplant recipients, and the importance of a histopathologic diagnosis and vigilant follow-up of such lesions.

Detection of the Epstein–Barr Virus (EBV) by In situ Hybridization as Definitive Diagnosis of Hairy Leukoplakia

Histopathological findings in cases of hairy leukoplakia (HL) are not exclusive to this lesion. A total of 36 tissue samples from patients previously diagnosed with HL based solely on morphological aspects were used in this study. Our purpose was to confirm the presence of Epstein-Barr virus (EBV) in these tissue samples by in situ hybridization (ISH), and to compare the detection of EBV with specific histopathological findings observed in each case. Among the 36 specimens, 80.55% were EBV positive, confirming the previous clinical and histhophatological diagnosis. None of the histopathological findings analyzed correlated with the presence or absence of EBV. This shows that a definitive diagnosis of HL cannot be established based on histopathological findings alone. Because there are many important implications on the establishment of definitive diagnosis of HL, the detection of EBV by ISH is obligatory.

Absence of Epstein-Barr virus in olfactory neuroblastoma

The sinonasal region is a frequent site for Epstein-Barr virus (EBV) related tumours, including nasopharyngeal carcinoma, sinonasal undifferentiated carcinoma and malignant lymphoma. Olfactory neuroblastoma is a rare neoplasm, arises from olfactory epithelium, is found in the upper nasal cavity, and may be confused morphologically with the above tumours. Our aim was to determine whether EBV is associated with olfactory neuroblastoma. We collected tissue samples for 16 olfactory neuroblastomas over an 11-year period at Kaohsiung Medical University Hospital. Diagnoses were confirmed from pathological review and immunohistochemistry. We used in situ hybridisation for EBV encoded small RNAs in those tumours. None of the epithelial cells in the 16 cases of olfactory neuroblastoma was positive for EBV. EBV does not play a major role in the aetiology of olfactory neuroblastoma.

Lymphoepithelioma-like carcinoma of the uterine cervix: absence of Epstein-Barr virus and high-risk human papilloma virus infection

Lymphoepithelioma-like carcinoma (LELC) is a rare variant of squamous cell carcinoma of the uterine cervix. In contrast to Asian women, Caucasian women have a low prevalence of cervical LELC with no infection of the Epstein-Barr virus (EBV) and sporadic appearances of human papilloma viruses (HPV) in Caucasian women. We report a 36-year-old female Caucasian patient with LELC of the cervix. In contrast to the present studies, our case of LELC of the cervix is not associated with EBV or high-risk HPV appearances. This is the first study, that showed no relationship between LELC of the uterine cervix with EBV and high-risk HPV infections in a Caucasian woman.

Absence of Epstein-Barr Virus, Human Papillomavirus, and Simian Virus 40 in Patients of Central European Origin With Lymphoepithelioma-like Carcinoma of the Skin

The authors report 10 cases of lymphoepithelioma-like carcinoma (LELC) of the skin and the results of a molecular biological study for HPV, EBV, and SV40 in lesional tissues. All patients originated from Central Europe. There were seven men and three women, ranging in age from 57 to 86 years. Locations included the face (n = 4), scalp (n = 2), penis (n = 2), and retroauricular area (n = 1); location was unknown for one subject. All but two patients presented with a tumor confined to the skin; in both patients with the penile carcinoma, the tumors had metastasized to an inguinal lymph node. Six patients with available follow-up included four individuals with no evidence of tumor metastasis or recurrences at 2, 3, 4, and 5 years, one patient who died with metastatic disease 7 years after diagnosis, and one patient who died of an unrelated course. Microscopically, all cases showed distinctive features of LELC characterized by variably sized and shaped nodules or syncytial sheets of epithelial cells that contained vesicular chromatic and prominent nucleoli and that were permeated and surrounded by small, well-differentiated lymphocytes and plasma cells. Because all 10 cases studied proved negative for EBV, HPV, and SV40, these viruses seem to play no causal role in LELC of the skin in patients from Central Europe.

Benign, atypical and malignant lymphoproliferative disorders in rheumatoid arthritis patients

Lymphadenopathy, which may be associated with systemic symptoms, is frequently associated with rheumatoid arthritis (RA). Reactive non-neoplastic tissue comprises the majority of the lymph node lesions. However, several cohort studies have demonstrated that RA has an increased risk of non-Hodgkin's lymphomas (NHLs). Since the early 1990s, an atypical or malignant lymphoproliferative disorders (LPD) in patients immunosupressed with methtorexate (MTX) therapy for RA has been emphasized, namely MTX-associated LPDs. Epstein–Barr virus (EBV) has received attention in connection with the etiology of RA. The present review describes the clinicopathologic and immunohistochemical findings of reactive, atypical and malignant LPDs associated with RA along with the presence or absence of EBV in LPDs using the in situ hybridization (ISH) method. The majority of reactive lymph node lesions exhibit reactive follicular hyperplasia with interfollicular polyclonal plasmacytosis. Atypical LPDs rarely appears in RA patients. However, these cases occasionally pose difficult problems in the differential diagnosis from malignant lymphomas associated with RA or atypical and malignant LPDs showing RA-like clinicopathological findings. Clinicopathologically, three types of atypical LPDs have delineated, i.e. (i) resembling multicentric Castleman’s disease (MCD); (ii) atypical paracortical hyperplasia with lymphoid follicles (APHLF) and; (iii) atypical lymphoplasmacytic immunoblastic proliferation. Malignant lymphoma associated with RA is characterized by; (i) predominance of elderly cases; (ii) usually female predominance, reflecting the sex ratio of RA; (iii) longstanding history of RA; (iv) relatively frequent advanced stage of disease; (v) majority of the patients had the B-cell phenotype; and (vi) an increased frequency of diffuse large B-cell lymphoma (DLBCL) in RA. It is unlikely that EBV is the causative agent of either reactive or atypical LPD. Among malignant lymphomas, EBV-associated lymphoma comprised only a small fraction of all NHLs in the general RA patient population.