The relationship between atypical lymphocytosis and serological tests in infectious mononucleosis

The relationship between serological tests for infectious mononucleosis was compared with atypical lymphocytosis in one hundred cases with clinical features of the disease. All blood samples showed atypical lymphocytosis greater than 20 per cent of the total leucocyte count. Positive serological tests were obtained in 69 of these patients, whereas in patients with more than 40 per cent atypical lymphocytosis all had positive serology. Of the 31 patients with negative serological tests for infectious mononucleosis, significant antibody titres to Toxoplasma gondii were obtained in five and to cytomegalovirus in two. Twenty-four patients remained undiagnosed. The Paul-Bunnell-Davidsohn positive patients had a significantly higher white cell count, lymphocyte count and atypical lymphocyte count than the toxoplasmosis group or the undiagnosed group.     

Effect of Epstein-Barr virus infection on response to chemotherapy and survival in Hodgkin's disease.

We have analyzed paraffin sections from 190 patients with histologically confirmed Hodgkin's disease (HD) for the presence of Epstein-Barr virus (EBV) using in situ hybridization to detect the EBV-encoded Epstein-Barr virus early RNAs (EBERs) and immunohistochemistry to identify latent membrane protein-1 (LMP1) expression. EBV was present in the tumor cells in 51 HD cases (27%) and was mainly confined to the mixed cellularity and nodular sclerosis subtypes. There was no difference between EBV-positive and EBV-negative HD patients with regard to age, clinical stage, presentation, and the number of alternating chemotherapy cycles of ChIVPP and PABIOE received. The complete remission rate after study chemotherapy was 80% in EBV-positive patients versus 69% in EBV-negative patients (P =.05). The 2-year failure-free survival rate was significantly better for EBV-positive patients when compared with the EBV-negative HD group (P =.02). Although 2-year and 5-year overall survival rates were better for EBV-positive HD patients, the differences were not statistically significant (P =.18 and P =.40, respectively). In conclusion, the results confirm the favorable prognostic value of EBV in the tumor cells of HD patients and suggest important differences in response to chemotherapy between EBV-positive and EBV-negative patients.     

Heterogeneity of risk factors and antibody profiles in epstein-barr virus genome-positive and -negative hodgkin lymphoma

BACKGROUND: Hodgkin lymphoma (HL) tumors that contain the Epstein-Barr virus (EBV) genome may differ etiologically from EBV-negative HL tumors. METHODS: A case-case study examining heterogeneity of risk factors between disease subgroups compared personal characteristics and EBV antibodies between 95 EBV-positive and 303 EBV-negative patients with HL. RESULTS: We confirmed previous associations of EBV-positive HL with older age, male sex, and mixed-cellularity (MC) histological subtypes. EBV-positive patients were less educated and more likely to have smoked cigarettes and had more prevalent and higher EBV antibody titers, compared with EBV-negative patients. After adjustment for all independent risk factors, those most strongly associated with EBV-positive HL were histological subtypes (odds ratio [OR] for MC vs. nodular sclerosis histology, 3.2; 95% confidence interval [CI], 1.4-7.2), elevated anti-viral capsid antigen level (OR, 3.1; 95% CI, 1.6-6.0), and less education (OR, 0.7; 95% CI, 0.5-1.0). Cigarette smoking and a low anti-Epstein-Barr nuclear protein (EBNA) 1 : anti-EBNA-2 ratio were also marginally associated with EBV-positive HL. CONCLUSIONS: EBV-positive HL is more common among individuals who have markers of diminished cellular immunity and an abnormal EBV antibody response. EBV appears to participate in the etiology of EBV-positive HL but may not be involved in EBV-negative HL.     

Impact of tumor Epstein-Barr virus status on presenting features and outcome in age-defined subgroups of patients with classic Hodgkin lymphoma: a population-based study

The association between tumor Epstein-Barr virus (EBV) status and clinical outcome in Hodgkin lymphoma (HL) is controversial. This population-based study assessed the impact of EBV status on survival in age-stratified cohorts of adults with classic HL (cHL). Data from 437 cases were analyzed with a median follow-up of 93 months. Overall survival (OS) was significantly better for EBV-negative compared with EBV-positive patients (P < .001), with 5-year survival rates of 81% and 66%, respectively; disease-specific survival (DSS) was also greater for EBV-negative patients (P = .03). The impact of EBV status varied with age at diagnosis. In patients aged 16 to 34 years, EBV-associated cases had a survival advantage compared with EBV-negative cases, but differences were not statistically significant (P = .21). Among patients 50 years or older, EBV positivity was associated with a significantly poorer outcome (P = .003). Excess deaths occurred in EBV-positive patients with both early- and advanced-stage disease. In multivariate analysis of OS in the older patients, EBV status retained statistical significance after adjusting for the effects of sex, stage, and B symptoms (P = .01). Impaired immune status may contribute to the development of EBV-positive cHL in older patients, and strategies aimed at boosting the immune response should be investigated in the treatment of these patients.     

Serum levels of lymphokines and soluble cellular receptors in primary Epstein-Barr virus infection and in patients with chronic fatigue syndrome.

The immunopathology in primary Epstein-Barr virus (EBV) infections and in chronic fatigue syndrome was studied by examining serum levels of interleukins (IL) and of soluble T cell receptors in serum samples. Serum samples were from patients during and 6 months after primary EBV-induced infectious mononucleosis and from patients with chronic fatigue syndrome and serologic evidence of EBV reactivation. Markers for T lymphocyte activation (soluble IL-2 and CD8) and for monocyte activation (neopterin) were significantly elevated during acute infectious mononucleosis but not in patients with chronic fatigue syndrome. Interferon-alpha, IL-1 beta, and IL-6 levels were not significantly increased in any patient group but inferferon-gamma levels were significantly increased during the acute phase of infectious mononucleosis. The levels of IL-1 alpha were significantly higher than in controls both in patients with infectious mononucleosis and in those with chronic fatigue syndrome. In the latter, the lack of most markers for lymphocyte activation found in patients with infectious mononucleosis makes it less likely that EBV reactivation causes symptoms.     

High prevalence of Epstein-Barr virus in gastric remnant carcinoma after Billroth-II reconstruction

BACKGROUND: Epstein-Barr virus (EBV) has been detected in about 10% of gastric carcinoma cases worldwide, and a high prevalence of EBV involvement in gastric remnant carcinoma has been reported recently. Details of the background remnant stomach of EBV-positive lesions, however, have not been well clarified. METHODS: We screened 17 consecutive gastric remnant carcinoma lesions resected surgically. To detect EBV, we used in situ hybridization (ISH) for EBV-encoded small RNA1 (EBER-1) and we compared the clinicopathologic feature between EBV-positive and -negative gastric remnant carcinoma cases. RESULTS: EBV was detected in 41.8% (7 of 17) of the lesions by EBER-1 ISH. All 7 EBV-positive lesions developed in the anastomotic site had undergone Billroth-II reconstruction excess 20 years previously (mean 26.4 years). Histologically, all EBV-positive lesions were poorly differentiated adenocarcinomas with intense lymphocyte infiltration. In the adjacent mucosa of carcinomas, moderate or marked intestinal metaplasia was found in 85.7% (6 of 7) of EBV-positive lesions and in 40% (4 of 10) of EBV-negative lesions. CONCLUSIONS: EBV infection is strongly associated with gastric remnant carcinoma. Atrophic change of remnant gastritis in Billroth-II anastomoses is considered to be the carcinogenic background for EBV-positive gastric remnant carcinoma.     

胃癌患者EB病毒感染与c-erbB-2基因表达的研究

目的研究胃癌患者癌组织中EB病毒(Epstein-Barrvirus,EBV)感染与c-erbB-2表达的关系。方法原位杂交法检测胃癌患者石蜡标本中EBV编码的小RNA(EBER1);免疫组化法检测EBV阳性胃癌(EBVaGC)及EBV阴性胃癌(EB-VnGC)标本C-erbB-2蛋白的表达状况。结果217例胃癌患者癌组织标本中检测到23例EBER1阳性;EBVaGC组C-erbB-2蛋白表达显著高于EBVnGC组(P<0.05)。结论EBV感染可能上调了癌基因c-erbB-2表达。     

GI Cancer High Prevalence of Epstein-Barr Virus in Gastric Remnant Carcinoma after Billroth-II Reconstruction

Background : Epstein-Barr virus (EBV) has been detected in about 10% of gastric carcinoma cases worldwide, and a high prevalence of EBV involvement in gastric remnant carcinoma has been reported recently. Details of the background remnant stomach of EBV-positive lesions, however, have not been well clarified. Methods : We screened 17 consecutive gastric remnant carcinoma lesions resected surgically. To detect EBV, we used in situ hybridization (ISH) for EBV-encoded small RNA1 (EBER-1) and we compared the clinicopathologic feature between EBV-positive and-negative gastric remnant carcinoma cases. Results : EBV was detected in 41.8% (7 of 17) of the lesions by EBER-1 ISH. All 7 EBVpositive lesions developed in the anastomotic site had undergone Billroth-II reconstruction excess 20 years previously (mean 26.4 years). Histologically, all EBV-positive lesions were poorly differentiated adenocarcinomas with intense lymphocyte infiltration. In the adjacent mucosa of carcinomas, moderate or marked intestinal metaplasia was found in 85.7% (6 of 7) of EBV-positive lesions and in 40% (4 of 10) of EBV-negative lesions. Conclusions : EBV infection is strongly associated with gastric remnant carcinoma. Atrophic change of remnant gastritis in Billroth-II anastomoses is considered to be the carcinogenic background for EBV-positive gastric remnant carcinoma.     

EB病毒感染与系统性红斑狼疮患者外周血中B淋巴细胞异常活化的相关性研究

目的 研究EB病毒(EBV)感染与系统性红斑狼疮(SLE)患者外周血B淋巴细胞异常活化及疾病活动性的关系,探讨EBV在SLE病因学研究中的意义.方法 应用聚合酶链反应(PCR)-Southern杂交技术检测52例SLE患者和23名健康对照组外周血中EBV特异性BamH I-W基因片段;同时应用流式细胞术检测患者及对照组外周血中CD19+、CD23+/CD19+B淋巴细胞的表达情况.结果 ①PCR-Southern杂交检测结果显示SLE患者组EBV特异性BamH I-W片段阳性率明显高于健康对照组(P<0.01).②活动期SLE患者组CD23+/CD19+双阳性细胞的表达率与稳定期及健康对照组比较差异均有统计学意义(P<0.01).③EBV-DNA阳性患者组CD19+及CD23+/CD19+细胞的表达率明显高于EBV-DNA阴性SLE组(P<0.05);EBV-DNA阳性患者组CD19+B淋巴细胞表达CD23的平均荧光强度明显高于E-BV-DNA阴性患者组(P<0.01).结论 SLE患者体内EBV感染与B淋巴细胞异常活化有一定的相关关系,EBV可能通过上调B淋巴细胞持续高水平表达CD23从而感染并活化B淋巴细胞;EBV与SLE的发生发展有一定关系.     

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

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

Bronchoalveolar lavage cell data in amiodarone-associated pneumonitis. Evaluation in 22 patients.

To assess the value of bronchoalveolar lavage (BAL) for diagnosis, understanding, and treatment of amiodarone-associated pneumonitis, we examined the results of BAL total and differential cell counts and phenotyping of lymphocytes in 22 patients with this lung disorder and in 33 normal subjects. Overall, the total cell count was found to be almost the same as that seen in control subjects; the macrophage population was significantly reduced, and the lymphocyte, neutrophil, and eosinophil populations were increased in absolute number and percentage. When results were analyzed individually, BAL data appeared to be distributed according to two patterns. In the first pattern, there was no abnormal lymphocytosis. In the second pattern a lymphocyte alveolitis was found in percentage and in absolute number. This lymphocyte alveolitis was present either alone or associated with neutrophil alveolitis or with eosinophil alveolitis. In the first pattern, despite the normal level of the lymphocyte population, the percentage of CD4 T-lymphocytes and the CD4:CD8 T-lymphocyte ratio were significantly lowered. In the second pattern the CD8 T-lymphocyte count was increased in absolute number and percentage, with a low CD4:CD8 ratio. In six patients relavaged two to four months after amiodarone withdrawal, there was a significant fall in alveolar lymphocytosis, but the progressive increase in the neutrophil population over time seemed to be associated with the seriousness and progression of the disease. Finally, these findings closely resembled those obtained in patients with hypersensitivity pneumonitis due to inhalation of organic dust and suggest that an underlying immunologic cell-mediated mechanism may play a role in this iatrogenic pulmonary disease.     

T cell leukemia I oncogene expression depends on the presence of Epstein-Barr virus in the virus-carrying Burkitt lymphoma lines

We used a modified subtractive suppression hybridization to identify cellular genes that show altered expression in Burkitt lymphomas (BLs) in the presence of Epstein-Barr virus (EBV). Comparison of the gene expression patterns of an EBV-negative clone of the originally EBV-positive BL line Akata, with its Neo(R)-EBV derivative, revealed a significant difference in the expression of the T cell leukemia 1 oncogene (TCL-1). Subsequent expression studies showed that the original EBV-positive Akata line and the EBV-reconstituted derivative expressed high levels of TCL-1, whereas the EBV-negative variant showed only a low level of expression. Two other independently established EBV-positive BLs (Mutu and OMA) that have also thrown off EBV showed a similar decrease in TCL-1 expression after virus loss. Reinfection with Neo(R)-EBV restored the TCL-1 expression levels in the EBV loss variants to as high a level as the originally EBV-positive lines. High-resolution immunostaining showed that TCL-1 was localized in both the cytoplasm and the nucleus. Our findings suggest that high expression of TCL-1 is necessary for the development of the BL phenotype. In view of the fact that germinal center B cells, regarded as the progenitors of BL, do not express TCL-1, we suggest that constitutive expression of this oncogene occurs by genetic or epigenetic changes in the EBV-negative BLs. In the originally EBV-positive BLs, the ability of the virus to switch on TCL-1 expression would obviate this need.     

Human leukocyte antigen class II alleles may contribute to the severity of hepatitis C virus-related liver disease

To investigate whether genetic differences in cytokine promoter polymorphisms effect various outcomes after exposure to Epstein-Barr virus (EBV) infection, 30 patients with EBV-positive gastric carcinoma (GC), 120 patients with EBV-negative GC, and 220 control subjects were enrolled. Promoter polymorphisms of tumor necrosis factor (TNF)-alpha at positions -238 and -308 and of interleukin (IL)-10 at position -1082 were determined. The frequency of the high-producer allele (-308A) in the TNF-alpha gene was significantly higher among EBV-positive GC patients compared with control subjects (23.3% vs. 12.0%, P<.05), whereas the frequency of the high-producer allele (-1082G) in the IL-10 gene was significantly higher among EBV-negative GC patients compared with control subjects (6.3% vs. 3.0%, P<.05). These data support the notion that genetic factors may modify the outcomes of infectious diseases through different TNF-alpha- or IL-10-producing capabilities.     

Regulation of transferrin receptors by iron in human erythroblasts

Previous reports of patients with persistent polyclonal B lymphocytosis have found associations with female sex, cigarette smoking, HLA-DR7 phenotype, and moderate elevation of peripheral blood polyclonal B lymphocytes. The presence of binucleated cells and atypical lymphocytes in the peripheral blood of these patients was highly suggestive of a viral infection, such as with the Epstein-Barr virus. We report a 47-year-old asymptomatic woman who was incidentally found to have persistent polyclonal B lymphocytosis and serum IgG against virus capsid antigen (VCA) and Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA) of EBV. The presence of EBV was investigated in the peripheral blood lymphocytes from this patient by in situ hybridization and polymerase chain reaction methods. EBV DNA was demonstrated in the lymphocyte fraction by polymerase chain reaction, and it was further located in lymphoid cells by in situ hybridization. These results indicate that persistent polyclonal B lymphocytosis is strongly associated with EBV.     

Quantification of circulating Epstein-Barr virus (EBV) DNA in the diagnosis and monitoring of natural killer cell and EBV-positive lymphomas in immunocompetent patients

In Epstein-Barr-virus (EBV)-positive lymphomas in immunocompetent patients, release of EBV DNA from tumor cells into the plasma might be useful for disease monitoring and prognostication. To test this hypothesis, we quantified serially plasma EBV DNA by quantitative polymerase chain reaction in 39 cases of EBV-positive (natural killer [NK] cell, n = 23; T cell, n = 8; B cell, n = 4; Hodgkin, n = 4) lymphomas. As control, EBV DNA was undetectable in 34 cases of EBV-negative lymphomas at diagnosis and during chemotherapy. In all cases of EBV-positive lymphomas, EBV DNA was detectable (10(5)-10(10) copies/mL) at diagnosis. It paralleled the clinical course, with EBV DNA becoming undetectable at remission and remaining elevated in refractory disease. On multivariate analysis, high-presentation EBV DNA (> 7.3 x 10(7) copies/mL) was significantly associated with an inferior overall survival (OS). Subgroup analysis of NK cell lymphomas, the largest cohort in this study, showed that presentation EBV DNA was correlated with disease stage and lactate dehydrogenase. On multivariate analysis, high-presentation EBV DNA (> 6.1 x 10(7) copies/mL) was significantly associated with an inferior disease-free survival. During treatment, patients with EBV DNA that showed further increases or failed to become undetectable had significantly inferior OS. In EBV-positive lymphomas, plasma EBV DNA is valuable as a tumor biomarker and for prognostication.     

Persistent lymphocytosis of natural killer cells in autoimmune thrombocytopenic purpura (ATP) patients after splenectomy

We report three patients with primary autoimmune thrombocytopenic purpura (ATP) who developed an absolute lymphocytosis (lymphocyte count > 5×10⁹/I) after splenectomy and with a lymphocyte count between 5.4 and 8.9×10⁹/I. An immunophenotype study showed that the peripheral blood lymphocytosis was a persistent NK cell expansion (CD2⁺, CD56⁺, CD3^-), and was characterized by a typical large granular lymphocytes (LGL) morphology. Two of these three ATP patients were refractory to splenectomy.     

Association between the Epstein-Barr virus and Hodgkin's lymphoma in the North-Eastern part of Hungary: effects on therapy and survival

This retrospective study included 109 patients with Hodgkin's lymphoma (HL; 45 females, 64 males). In 47 of the 109 HL patients (43%), immunohistochemical analysis of their formalin-fixed, paraffin-embedded histologic samples revealed Epstein-Barr virus (EBV) by latent membrane protein (LMP) 1. The highest virus association (50%) was found with the mixed cellularity histologic subtype, especially in patients aged 11-20 and >50 years. Virus positivity in nodular sclerosis was 35% (negative cases accumulated in patients aged 15-30 years). Regarding clinical stages, histologic subtypes, general symptoms, treatments employed and response to treatment, the EBV-positive group was not significantly different from the virus-negative group. During the mean follow-up time of 83 months (9-300 months), the overall or event-free survival of EBV-negative patients was more favorable than that of EBV-positive patients, although the difference was not significant (p = 0.16 and p = 0.24, respectively). EBV infection may be involved in the pathogenesis of HL in our Hungarian study cohort, but it does not significantly affect clinical symptoms, therapeutic results or complete and event-free survival of HL patients.     

Epstein-Barr virus-associated gastric carcinomas: relation to H. pylori infection and genetic alterations

BACKGROUND & AIMS: The association of Epstein-Barr virus (EBV) and gastric carcinomas (GCs) has been shown to vary among different populations and certain histological subtypes. Few studies have addressed the status of Helicobacter pylori infection and genetic alterations in these EBV-positive or -negative GCs. METHODS: Eleven gastric lymphoepithelioma-like carcinomas (LELCs) and 139 cases of common non-LELCs were evaluated for the presence of EBV DNA using polymerase chain reaction (PCR) and RNA in situ hybridization. H. pylori infection was determined by anti-H. pylori immunoglobulin G in preoperative sera. Immunostaining for p53, c-erbB-2, and E-cadherin was performed. Microsatellite instability was analyzed by PCR using 10 primers. RESULTS: EBV was detected in 11 (100%) LELCs and in 19 (13.7%) of 139 common GCs. Compared with EBV-negative GCs, gastric LELCs tended to have a relatively higher frequency of proximal location, diffuse histological subtype, p53 overexpression, and reduced E-cadherin expression but a lower frequency of lymph node metastasis, previous H. pylori infection, and c-erbB-2 overexpression. In contrast, no significant difference of clinicopathologic and genetic profiles was observed between EBV-positive non-LELC GCs and EBV-negative GCs. No correlation of microsatellite instability was found among these 3 subsets of GCs. CONCLUSIONS: Dissecting clinicopathologic characteristics and infection status of EBV and H. pylori provide additional evidence of etiological and genetic heterogeneity for GC. Distinct clinicopathologic and genetic pathways exist in gastric LELCs, in which EBV may play a more important role than H. pylori infection.     

Cellular expressions and serum concentrations of Fas ligand and Fas receptor in patients with infectious mononucleosis

Although lymphocytosis and neutropenia are commonly associated with infectious mononucleosis (IM), the precise mechanisms involved remain unclear. Accumulated evidence has revealed that the apoptosis-mediating system, Fas receptor/Fas ligand (Fas-R/Fas-L), plays an important role in this disease. Recently, lymphocytes, monocytes, and neutrophils have been reported to constitutively express Fas-R, and the Epstein-Barr virus (EBV) has been shown to activate, in addition to B cells, peripheral blood CD8+ T cells, monocytes, and neutrophils. We elucidated cell surface expression and serum concentrations of Fas-R and Fas-L in patients with IM in an effort to more clearly define the role and contribution of apoptosis in this disease. The expression of lymphocyte surface Fas-L and Fas-R was significantly increased in patients with IM (P < .005 and P < .001, respectively), and among lymphocytes, CD4+ or CD8+ populations contained Fas-R+ as well as Fas-R- subpopulations. The constitutive Fas-R expression levels of monocytes and neutrophils were also increased in IM. Moreover, serum levels of both soluble Fas-L and Fas-R were significantly higher in patients with IM than in healthy volunteers (P < .001 and P < .0001, respectively). Positive relationships between the number of peripheral blood CD95+ lymphocytes and white blood cell count, number of lymphocytes, or number of CD4+ or CD8+ lymphocytes were observed. Our results suggest that the Fas-R/Fas-L system might play a role in eliminating EBV-infected or -activated peripheral blood cells by cell-to-cell contact or in an autocrine and/or paracrine fashion in patients with IM.