De novo infection of hepatitis B virus in patients with orthotopic liver transplantation: analysis by determining complete sequence of the genome

De novo infection of hepatitis B virus (HBV) occurs after liver transplantation from donors with HBV markers that suggest past infection. In the present study, the complete nucleotide sequences of HBV derived from a donor and recipients were determined to determine the clinical and virological characteristics. A total of 57 donor-recipient pairs, which underwent living-related orthotopic liver transplantation, were enrolled in the present study; all were negative for HBsAg before transplantation. HBV DNA was tested in serum, liver tissue, and peripheral blood mononuclear cells (PBMCs) by the polymerase chain reaction (PCR). The nucleotide sequence of HBV was determined based on PCR products and the phylogenetic analysis. De novo infection of HBV was found in 3 of the 57 recipients. Anti-HBc was positive in all donors of 3 recipients with the de novo infection but was positive only in 4 donors of the remaining 54 recipients (P=0.001). HBV DNA was detected in the liver but not in the serum or PBMCs in donor 3 whose recipient developed de novo HBV infection. The nucleotide sequence covering entire genome of HBV (3,215 bases) derived from the liver of donor 3 had a homology of 99.8-100% with that derived from the serum of corresponding recipient 3. The strain of recipient 3 showed the closest association with that of the donor 3 by phylogenetic analysis. Complete sequences from two recipients with de novo HBV infection including recipient 3 conserved the basic organisation of HBV genome. Analysis of the entire nucleotide sequence of HBV genome proved that HBV existed in the liver of the donor with anti-HBc, and it caused de novo infection in the corresponding recipient.     

Adoptive cellular immunotherapy for EBV lymphoproliferative diseases

Summary: Reactivation of EBV (Epstein-Barr virus) after bone marrow transplantation can result in EBV-associated lymphoproliferative disease (EBV-LPD), We have administered donor-derived EBV-specific cytotoxic T lymphocytes (CTL) to patients who are at high risk of this complication after receiving a T-cell-depleted allograft from a matched unrelated or mismatched related donor. The cells were marked with (he neo GENE U fore infusion so that we could evaluate their persistence and efficacy, CTL infusion produced a virus-specific immune response to EBV that persisted for up to 2 years. None of the 36 patients who received prophylactic CTLs have developed EBV-LPD compared with a cumulative risk of 14% in patients who did not receive this treatment. Strong evidence of clinically valuable immune activity conies from 6 of these 36 patients whose pre-CTL levels of EBV DNA were elevated to a degree strongly predictive of the onset of lymphoma. In each of these cases, the levels returned to baseline after CTL infusion. 2 patients who were treated for clinically evident EBV-LPD attained prolonged remission after CTL infusion and in situ hybridization and semiquantitative PCR showed that the gene-marked CTL had selectively accumulated at disease sites, The prophylactic CTL treatment lacked acute adverse effects, whereas 1 patient who received CTLs for bulky established disease developed initial tumor swelling and respiratory obstruction. We conclude that EBV-specific CTLs are a safe and effective prophylaxis for EBV lymphoma and can also eradicate established disease. This approach is now being extended lo other viruses that produce post-trans-plant morbidity and to other EBV-associated malignancies.     

A multiplex calibrated real-time PCR assay for quantitation of DNA of EBV-1 and 2

Accurate and highly sensitive tests for the diagnosis of active Epstein-Barr virus (EBV) infection are essential for the clinical management of individuals infected with EBV. A calibrated quantitative real-time PCR assay for the measurement of EBV DNA of both EBV-1 and 2 subtypes was developed, combining the detection of the EBV DNA and a synthetic DNA calibrator in a multiplex PCR format. The assay displays a wide dynamic range and a high degree of accuracy even in the presence of 1μg of human genomic DNA. This assay measures with the same efficiency EBV DNA from strains prevalent in different geographic areas. The clinical sensitivity and specificity of the system were evaluated by testing 181 peripheral blood mononuclear cell (PBMCs) and plasma specimens obtained from 21 patients subjected to bone marrow transplantation, 70 HIV-seropositive subjects and 23 healthy controls. Patients affected by EBV-associated post-transplant lymphoprolipherative disorders had the highest frequency of EBV detection and the highest viral load. Persons infected with HIV had higher levels of EBV DNA load in PBMCs and a higher frequency of EBV plasma viremia compared to healthy controls. In conclusion, this new assay provides a reliable high-throughput method for the quantitation of EBV DNA in clinical samples.     

High fatality rate of Epstein-Barr virus-associated lymphoproliferative disorder occurring after bone marrow transplantation with rabbit antithymocyte globulin conditioning regimens

Epstein-Barr virus (EBV)-associated lymphoproliferative disorder (EBV-LPD) following bone marrow transplantation can be fatal. The major risk factors for the development of EBV-LPD are ex vivo T-cell depletion or in vivo T-cell depletion with either antithymocyte globulin (ATG) or monoclonal anti-T-cell antibodies. Between March 1999 and January 2001, a total of 23 transplants with ATG of equine source (20 transplants) and ATG of rabbit source (3 transplants) used as part of the preparatory regimen were performed at the Barbara Ann Karmanos Cancer Institute in Detroit, Mich. The three patients who received rabbit ATG developed EBV-LPD between 60 and 90 days following bone marrow transplantation. However, there were no cases of EBV-LPD in the equine group. Treatment given in these cases consisted of tapering immunosuppression, antiviral therapy, unprocessed donor lymphocyte infusion, mobilized peripheral blood progenitor cell rescue infusion (one patient), and chemotherapy (one patient). All three patients died of complications from EBV-LPD. The association of rabbit ATG with the development of EBV-LPD suggests that patients receiving rabbit ATG as part of their preparatory regimens require close monitoring of the EBV viral load and possible early intervention with antiviral therapy.     

Assessment of automated DNA extraction coupled with real-time PCR for measuring Epstein-Barr virus load in whole blood, peripheral mononuclear cells and plasma.

BACKGROUND: Epstein-Barr virus (EBV) DNA load monitoring in blood has been shown to be essential for the diagnosis of EBV-associated diseases. However, the methods currently used to assess EBV DNA load are often time-consuming and require prior blood separation. OBJECTIVES: The aim of this study was to evaluate the relative diagnostic value of EBV DNA load monitoring in whole blood, peripheral blood mononuclear cells (PBMCs) and plasma after automated DNA extraction using the MagNA Pure extractor followed by LightCycler real-time quantitative PCR (LC-PCR). STUDY DESIGN: First, EBV DNA load was assessed retrospectively after automated or manual extraction on 104 PBMC specimens. Second, EBV DNA load was determined prospectively with the automated extraction procedure in the whole blood, PBMCs and plasma of 100 samples from patients with EBV-related diseases (group 1, n = 20), HIV-seropositive individuals (group 2, n = 66), and healthy EBV carriers (group 3, n = 14). RESULTS: A good correlation was observed between automated and manual extraction on 104 PBMC specimens (r = 0.956; P < 0.0001). In the prospective study, 67 samples were positive in both whole blood and PBMCs, with a good correlation between EBV DNA loads in whole blood and PBMCs (r = 0.936; P < 0.0001). Only 18/100 samples were positive in plasma. Higher viral loads were regularly observed in the three blood compartments from group 1 than from groups 2 and 3. CONCLUSION: This study demonstrated that an automated extraction of EBV DNA is easier to perform in whole blood or plasma than in PBMCs and facilitates the standardisation of EBV DNA measurement by real-time quantitative PCR. The quantitative detection of EBV DNA load in whole blood appeared more sensitive than in plasma for infectious mononucleosis in immunocompetent patients, probably because of a rapid loss of plasmatic EBV DNA. In transplant patients, EBV DNA load monitoring in whole blood and in plasma turned out to be equivalent in terms of feasibility and accuracy for the early diagnosis of post-transplant lymphoproliferative diseases (PTLDs).     

EB病毒感染及30 bp缺失潜伏膜蛋白1基因在鼻咽部癌不同组织学类型中的比较

目的 比较鼻咽部癌4种组织学类型中EB病毒的感染率以及野生型潜伏膜蛋白(LMP)1和缺失型LMPl EB病毒变异株单独或双重感染的检出频率,阐明缺失型LMPl基因在鼻咽癌变过程中的作用。方法 采用EBER原位杂交法检测117例鼻咽部癌,包括48例非角化性癌、25例角化性鳞状细胞癌、5例腺鳞癌、6例黏液表皮样癌和33例腺癌标本。采用巢式聚合酶链反应(PCR)法检测99例EBER阳性的癌组织和53个健康成人的外周血单个核细胞EB病毒LMPl基因。结果EBER原位杂交示,48例非角化性癌和25例角化性鳞状细胞癌的EB病毒感染率均为100％;而腺鳞癌和黏液表皮样癌的：EB病毒感染率为9／11,腺癌为51,5％(17／33)。阳性病例中大多数非角化性癌细胞呈EBER阳性,而在17例腺癌中仅见到少数EBER阳性肿瘤细胞。在非角化性癌中检测到单独缺失型LMPl：EB病毒变异株的百分率(85.4％,41／48)不但高于健康成人外周血单个核细胞(8．7％,4／46),而且高于角化性鳞状细胞癌(16．0％,4／25)。在角化性鳞状细胞癌中检出野生型LMPl和缺失型LMPl基因EB病毒的双重感染率(56．0％,14／25)高于非角化性癌的(12．5％,6／48)。在腺癌和健康成人单个核细胞中检出的大多数EB病毒为野生型LMPl和缺失型LMPl基因变异株同时并存。非角化性癌／角化性鳞状细胞癌、腺鳞癌／黏液表皮样癌和腺癌之间的：EB病毒感染率差异有显著性意义。非角化性癌和角化性鳞状细胞癌EB病毒的感染率相同,但非角化性癌中单独缺失型LMPl EB病毒变异株检出率远远大于角化性鳞状细胞癌,角化性鳞状细胞癌多数含有野生型LMPl和缺失型LMPl双重EB病毒变异株。结论由于分化导向不同而发生的鼻咽部癌不同组织学类型具有不同的EB病毒感染率。缺失型LMPl基因的EB病毒变异株可能在非角化性癌中呈选择性优势。     

Analysis of LMP1 variants of EBV in Southern Thailand: evidence for strain-associated T-cell tropism and pathogenicity.

BACKGROUND: The latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) has sequence heterogeneity. Some of the variants are associated with altered tumorigenic activity and show geographically specific localization. In Thailand, the EBV genome is frequently detected in circulating T cells of T-cell diseases. OBJECTIVE: To determine the role of EBV LMP1 variation in the genesis of T-cell diseases, we focused on virus factors and analyzed EBV strains in Thailand. STUDY DESIGN: EBV DNA was extracted from 18 healthy individuals and 45 patients with T-cell diseases in Southern Thailand and 30 healthy individuals in Central Thailand. By using PCR-direct sequencing method, nucleotide sequences corresponding to the carboxyl terminus of the LMP1 were determined. RESULTS: Four known strains, B95-8 prototype, China 1, China 2 and Mediterranean (Med) and two novel strains, Southeast Asia 1 (SEA 1) and Southeast Asia 2 (SEA 2) were identified. The prevalence of China 2 strain was significantly different (p=0.006) between Central and Southern Thailand. Higher prevalence (p=0.026) of 30-bp deletion type in the Southern Thais was observed. The LMP1 Med strain was associated with the worse prognosis (p=0.029). Among T-cell diseases patients, CD3(+)-cell oriented infection was recognized in SEA1 strain (p=0.025). CONCLUSION: The distribution of EBV strains may be associated with geographic/ethnic and clinical background in the Thai population. Certain EBV strains defined by their LMP1 sequence may influence cell tropism, disease association, or disease severity.     

Human herpesvirus 6 in human immunodeficiency virus-infected individuals: Association with early histologic phases of lymphadenopathy syndrome but not with malignant lymphoproliferative disorders

Preliminary evidence suggested that human herpesvirus-6 (HHV-6) may act as a cofactor in acquired immunodeficiency syndrome (AIDS) and may contribute to the pathogenesis of lymphoproliferative disorders occurring in individuals infected with the human immunodeficiency virus (HIV). To understand better the biological and clinical significance of HHV-6 infection in the context of HIV-related immunosuppression, the polymerase chain reaction was used to study the frequency and variant distribution of HHV-6 in peripheral blood mononucleated cells (PBMCs) from HIV-seropositive individuals, either asymptomatic or with lymphadenopathy syndrome (LAS) or with overt AIDS. Non-neoplastic and malignant lymphoproliferative disorders from both HIV-infected and HIV-seronegative patients were also investigated using the same series of samples for the presence of Epstein-Barr virus (EBV). When compared with healthy blood donors (12/42, 29%), HHV-6 prevalence in PBMCs showed a progressive decline in HIV-seropositive individuals with asymptomatic HIV infection (3/26, 11%) and in patients with LAS (1/13, 8%) and a significant reduction in patients with overt AIDS (1/20, 20%; P = 0.02). The decrease correlated with the number of CD4⁺ cells at the time of examination. In addition, HHV-6 DNA sequences were significantly more prevalent in LAS biopsies (13/20, 65%) than in HIV-unrelated reactive lymphadenopathies (2/10, 20%; P = 0.02) and the presence of HHV-6 sequences correlated closely with a histologic pattern of follicular hyperplasia (13/16, 81%; P = 0.003). Strikingly, HHV-6 prevalence decreased in PBMCs of LAS patients, suggesting that the likelihood of interactions between HHV-6 and HIV varies in different body districts. In particular, the demonstration that all HHV-6-carrying LAS samples were also positive for HIV infection suggests that LAS lymph nodes constitute one of the sites where biologically relevant interactions between the two viruses might occur. Also, the prevalence of EBV was higher in LAS (14/20, 70%) than in non-neoplastic lymph nodes from HIV-seronegative individuals (4/10, 40%), although the difference was not statistically significant. EBV was associated strongly with HIV-related malignant lymphoproliferative disorders, being detected in 100% of patients with Hodgkin's disease (HD) and 53% of B-cell non-Hodgkin's lymphomas (NHL). In contrast, the prevalence of HHV-6 DNA in HD and B-cell NHL arisen in HIV-infected patients (30% and 6%, respectively) was remarkably lower and similar to that observed in lymphoproliferative disorders from HIV-seronegative patients. Finally, as observed in healthy individuals, HHV-6 variant B was more prevalent than variant A in benign and malignant lymphoproliferative disorders from both HIV-infected and HIV-seronegative patients. These results suggest that the interactions between HHV-6 and HIV could be different in the various phases of HIV disease and in different districts; HHV-6 has probably no direct role in the pathogenesis of HIV-associated B-cell NHL and HD cases, and behave differently from EBV; and HIV-related immunosuppression does not alter the distribution of HHV-6 variants in these tissues, as observed in the case of EBV. © 1996 Wiley-Liss, Inc.     

Epstein-Barr virus and human papillomavirus infections and oropharyngeal squamous cell carcinomas

Epstein-Barr virus (EBV) and human papillomavirus (HPV) are known to exhibit oncogenic potential. Target cells for both viruses include oropharyngeal elements. The present study investigated whether EBV or HPV infection are associated with palatine tonsil carcinoma (PTC) and/or tongue carcinoma (TC). The study included 28 adult patients with oropharyngeal squamous cell carcinoma, including 14 patients with PTC and 14 patients with TC. The control group included 20 healthy adult volunteers. Sera of all patients and controls were tested for IgG anti-EA antibodies, IgG anti-VCA antibodies, and IgG anti-EBNA antibodies. DNA extracted from the tumors was tested for the presence of EBV DNAand HPV DNAusing PCR-ELISA. In parallel, the presence of EBV DNA was tested in the peripheral blood in all healthy individuals and patients. In addition, attempts were made to detect HPV 16 and HPV 18 using other PCR amplification techniques. Serum anti-EBV antibodies were detected in 24 patients (12 patients with PTC and 12 patients with TC). The frequency of detection of the antibodies did not significantly differ between the groups of patients and the control individuals. Most positive patients and controls demonstrated a serological pattern typical for past EBV infection. EBV DNA was identified in 12 cases of PTC and in 12 cases of TC (altogether 86% cases). In 10 PTC patients, 8 TC patients, and only 2 healthy individuals EBV DNA was detected in peripheral blood. HPV DNA was detected in only 3 cases (1 sample of PTC and 2 samples of TC). These results suggest that the etiopathogenesis of oropharyngeal cancers may be associated with EBV infection much more frequently than with HPV infection. Received: 26 March 2002 / Accepted: 23 September 2002 Correspondence to A. Szkaradkiewicz     

A monoclonal expansion of Epstein–Barr virus-infected natural killer cells after allogeneic peripheral blood stem cell transplantation

Here, we describe a Japanese woman showing a monoclonal expansion of EBV-infected natural killer (NK) cells after receiving allogeneic peripheral blood stem cell transplantation (PBSCT). The patient initially had T-cell-type chronic active EBV disease (CAEBV) and subsequently developed liver T-cell lymphoma. l-Asparaginase-containing chemotherapy led to a favorable lymphoma response. To eradicate CAEBV and the lymphoma, she further received allogeneic PBSCT from a human leukocyte antigen-matched sibling donor. After the PBSCT, the patient presented with transient lymphocytosis of NK cells, which were infected with a monoclonal EBV strain other than previously detected ones. These NK cells seemed to have been transmitted from the healthy donor to the recipient. The patient and donor remain well in spite of carrying these NK cells. This is the first report of an asymptomatic Japanese carrier harboring monoclonal EBV-infected NK cells.     

Specificities of IgE and IgG antibodies in patients with birch pollen allergy

58 sera from patients with established birch pollen allergy showed characteristic antibody-binding patterns in immunoblotting experiments. Regarding IgE, 56/58 patients recognized a protein of molecular weight (MW) 17 kilodaltons (kD), previously defined as Bet v I. 23/58 patients in addition reacted with a variety of 11 minor allergens with MWs ranging from 13 to 68 kD. A 13-kD protein was proved to represent an independent minor allergen. IgG binding in patients and healthy individuals was more pronounced on the minor allergens than on Bet v I. 3 different allergens were not detected by IgG of healthy individuals. In two-dimensional electrophoresis/immunoblot, a monoclonal antibody and human IgE (in both cases directed against Bet v I) detected a very similar cluster of spots, probably representing isoallergens of Bet v I.     

Human herpesvirus 6 (HHV-6) is transmitted from parent to child in an integrated form and characterization of cases with chromosomally integrated HHV-6 DNA

We obtained 7,566 peripheral blood mononuclear cell (PBMC) samples from 2,332 individuals and screened them for human herpesvirus infection. We identified five individuals who persistently harbored high copy numbers of human herpesvirus 6 (HHV-6) DNA in their PBMCs. HHV-6 DNA was also detected in other somatic tissues of these individuals. Five additional cases were identified among their family members. For two of these families, chromosomally integrated HHV-6 DNA (CIHHV-6) was detected in the PBMCs by fluorescence in situ hybridization. The prevalence of CIHHV-6 among all the subjects was 0.21%. The HHV-6 DNA was variant B in four families and variant A in one family. Antibodies to immediate early antigen and glycoprotein B were detected in 57 and 14% of individuals with CIHHV-6 and in 0 and 60% of healthy volunteers without CIHHV-6, respectively. HHV-6 could not be isolated from PBMCs with CIHHV-6. These cases shared no clinical features, and included three healthy individuals. Our data suggest that CIHHV-6 is rare but detectable in the general population and that hereditary transmission is one of the routes of HHV-6 transmission.     

Human herpesvirus 6 DNA in peripheral blood cells and saliva from immunocompetent individuals.

Human herpesvirus 6 (HHV-6) genome equivalents were quantitated in peripheral blood mononuclear cells (PBMCs) and saliva from 20 healthy individuals by the polymerase chain reaction (PCR). Nineteen of 20 subjects (95%) harbored HHV-6 DNA: 18 (90%) had HHV-6 in their PBMCs and 18 had HHV-6 in their saliva. Quantitative PCR revealed HHV-6 DNA levels ranging from negative to 4,000 HHV-6 genome equivalents per 10(6) PBMCs and from negative to 200,000 HHV-6 genome equivalents per ml of saliva. Longitudinal saliva samples from 15 HHV-6-seropositive subjects revealed salivary HHV-6 DNA persistence in 13 subjects. HHV-6 antibodies were detected in 17 of 19 subjects, with titers ranging from 1:400 to 1:51,200 (geometric mean titer, 1:2,500). Antibody titers did not correlate with HHV-6 DNA levels in PBMCs or saliva (P = 0.27 and P = 0.44, respectively). One subject with persistent HHV-6 DNA lacked detectable HHV-6 antibodies. The high prevalence of HHV-6 DNA in PBMCs and saliva supports the concept that HHV-6 exists at these sites in normal individuals.     

Detection of Epstein-Barr virus and human papillomavirus in head and neck tumors.

The presence of Epstein-Barr virus (EBV) DNA and human papillomavirus (HPV) DNA in 74 head and neck tumor tissues was examined by the polymerase chain reaction and DNA sequencing analysis. EBV DNA sequence was detected in all 30 nasopharyngeal-carcinoma tissue samples and in 30 of 44 other head and neck tumor samples. HPV DNA sequence was detected in 14 of 30 nasopharyngeal-carcinoma tissue samples and in 11 of 44 other tumor samples. Coinfection of both viruses was observed in 14 nasopharyngeal-carcinoma tissue samples but only in 5 other head and neck tumor samples including 3 hypopharyngeal-carcinoma tissue samples. Our data indicate that EBV is closely associated with nasopharyngeal- carcinoma and may also be related to hypopharyngeal-carcinoma. In addition, a relatively high percentage of EBV-positive nasopharyngeal- and hypopharyngeal-carcinoma tissue specimens contained HPV sequence. The significance of the coexistence of EBV and HPV in these tumor tissues requires further study.     

Cytomegalovirus,Epstein-Barr virus,and human herpesvirus-6 infections in patients with myalgic еncephalomyelitis/chronic fatigue syndrome

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a disabling multisystem chronic disease. The etiology and pathogenesis of ME/CFS are unknown. Infections of cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpesvirus-6 (HHV-6) are suspected as etiological agents for ME/CFS. This study aims to estimate prevalence and type (active/latent) of EBV, CMV, and HHV-6 infections in Bulgarian ME/CFS patients. In the study were included 58 patients with ME/CFS and 50 healthy controls. Virus-specific antibodies were detected by enzyme-linked immunosorbent assay and viral genomic sequences in peripheral blood mononuclear cell (PBMCs) and plasma samples by nested polymerase chain reaction (PCR). We did not observe any significant differences in virus-specific immunoglobulin G and immunoglobulin M positivity rates between patients with ME/CFS and control group. In ME/CFS plasma samples, EBV DNA was found in 24.1%, CMV DNA in 3.4%, and HHV-6 DNA in 1.7% of samples. EBV DNA was detected in 4%, and CMV and HHV-6 DNA were not found in plasma samples of controls. The frequency of viral genome detection in PBMCs of patients and controls was 74% vs 78% for CMV, 81% vs 84% for EBV, and 82.8% vs 82% for HHV-6. The difference in frequency of EBV active infection in ME/CFS and control group was statistically significant (P = .0027). No ME/CFS and control individuals with active CMV and HHV-6 infection were observed. In conclusion, this study using both serological and PCR-based techniques for distinguishing between active and latent infection showed high rate of active EBV infection among patients with ME/CFS indicating that at least in a subset of cases, EBV is important factor for the development of disease.     

Epstein-barr virus (EBV) in healthy carriers: Distribution of genotypes and 30 bp deletion in latent membrane protein-1 (LMP-1) oncogene

There are two types of Epstein Barr virus (EBV): EBV-1 and EBV-2, distinguished by genomic polymorphism in the genes encoding the nuclear antigens (EBNA-2, -3A, -3B, -3C). Latent membrane protein 1 (LMP-1) is an EBV protein with known oncogenic properties. Different variants had been described; among them, a 30 base pair (bp) deletion (del-LMP-1) had been reported in benign and malignant pathologies, but there is little information about its frequency in healthy populations. The aim of this study was to determine the distribution of the EBV genotypes and the 30 bp deletion frequency, in EBV healthy carriers from Argentina. Analysis of EBNA-3C and LMP-1 genes were done by polymerase chain reaction (PCR) followed by Southern blot hybridization on DNA of peripheral blood mononuclear cells (PBMCs) from blood bank donors. EBV-1 was present in 75.9% of samples, EBV-2 in 14.6%, and co-infections with both types in 6.5%. The deleted LMP-1 variant was found in 7.4% of analyzed samples, corresponding 3.2% to deleted variant alone and 4.2% to co-infections with non-deleted form. The non-deleted variant was found in 64.6% whereas in the remaining 28%, no PCR product was detected. These results showed that EBV-1 was the more prevalent type in healthy carriers of Argentina, similar to reports from others countries. A predominance of the non-deleted LMP-1 variant was observed. The presence of co-infections with both types and variants demonstrated that healthy individuals may also harbor multiple EBV infections.     

大鼠气管异位移植慢性排斥反应模型的建立及其与白细胞介素17的关系

背景：闭塞性细支气管炎是肺移植远期主要并发症之一,建立稳定可靠的动物模型是研究闭塞性细支气管炎的首要条件。 目的：建立大鼠气管异位移植慢性排斥反应模型,探讨白细胞介素17在该模型中的作用。 方法：按体质量配对,Wistar和SD大鼠随机分为2组(n=30)：Wistar(供体)→SD(受体)组和SD(供体)→SD(受体)组,颈前皮下组织包埋移植气管,不同时间点分别采样行组织病理学分析,对比观察移植气管上皮丢失、淋巴细胞计数、无核软骨细胞/软骨细胞总数及白细胞介素17表达量等指标。 结果与结论：两组受体全部存活。移植后第7,14,28天两组淋巴细胞计数对比差异均有显著性意义(P < 0.05);移植后第14,28天两组无核软骨细胞/软骨细胞总数比值相比差异均有显著性意义(P < 0.05);移植后第7,14,28天Wistar(供体)→SD(受体)组上皮丢失度分别为> 40%,100%,100%,SD(供体)→SD(受体)组未见明显丢失;移植后第28天Wistar(供体)→SD(受体)组闭塞性细支气管炎发生率为100%,SD(供体)→SD(受体)组未发生闭塞。移植后第28天Wistar(供体)→SD(受体)组白细胞介素17表达量显著高于SD(供体)→SD(受体)组(P < 0.05)。以上结果说明实验成功建立了Wistar(供体)→SD(受体)大鼠异位气管移植慢性排斥反应模型,为闭塞性细支气管炎研究提供了平台,白细胞介素17可能在其中发挥重要作用。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程      

鼻咽癌组织中人疱疹病毒6型感染及其临床意义

目的　探讨人疱疹病毒６ 型（ＨＨＶ６） 与鼻咽癌（ＮＰＣ） 发病的关系。方法　采用聚合酶链反应（ＰＣＲ） 和原位杂交同时检测正常鼻咽组织和ＮＰＣ 癌前鼻咽组织以及ＮＰＣ 组织中ＨＨＶ６ 和ＥＢＶＤＮＡ 的存在情况。采用免疫组化检测３６ 例ＮＰＣ 组织的ＥＢＶＬＭＰ１ 蛋白表达。结果　在４２ 例ＮＰＣ组织、２１ 例鼻咽癌前病变组织中,经ＰＣＲ检出ＨＨＶ６ ＤＮＡ,阳性率分别为３０－９％ （１３ 例） 和４－７ ％（１ 例） 。ＥＢＶＤＮＡ为９５－２％ （４０ 例） 和６６－６ ％ （１４ 例）。２７ 例正常鼻咽组织未检出ＨＨＶ６ ＤＮＡ,ＥＢＶＤＮＡ为２５－９％ （７ 例）。经ＩＳＨ,仅在１３ 例ＰＣＲＨＨＶ６ ＤＮＡ阳性的ＮＰＣ组织中检出１１ 例阳性,３６ 例ＮＰＣ组织、１０ 例鼻咽癌前病变组织ＩＳＨＥＢＶＤＮＡ 阳性。ＮＰＣ组织的ＥＢＶＬＭＰ１ 蛋白阳性率为４７－２ ％（１７３６）。结论　鼻咽癌组织中存在ＨＨＶ６ 感染,提示ＨＨＶ６ 感染与ＮＰＣ有关。ＨＨＶ６ 感染与ＥＢ病毒ＬＭＰ１ 蛋白表达上调呈正相关,提示在ＮＰＣ 的发生中ＨＨＶ６ 可能直接参与和（ 或） 通过上调ＥＢＶＬＭＰ１ 的表达而起一定作用     

EBNA-1 sequence variations reflect active EBV replication and disease status or quiescent latency in lymphocytes

EBV infects most of the global population, but only a small percentage of infected individuals develop EBV-associated malignancies. Host and viral factors may play a role in determining the clinical outcome. Because EBNA-1 functions as an oriP binding protein and links the episomal viral DNA to metaphase chromosomes, variation of EBNA-1 has been suggested to contribute to determining the tissue tropism of EBV and the development of various EBV-associated diseases. Five subtypes have been described, according to the amino acid at residue 487: P-ala (B95-8 prototype), P-thr (aa 487 ala to thr), V-val, V-pro and V-leu. Other studies, however, concluded that EBNA-1 sequence variation simply reflects the geographical distribution of EBV. To clarify these possibilities, we collected DNA samples from healthy individuals and patients with various EBV-associated diseases in Taiwan for PCR amplification and DNA sequencing. The results indicate that: 1) V-val EBNA-1 was detected in patients with nasopharyngeal carcinoma (NPC) and other EBV-associated malignant diseases; 2) the prototype P-ala strain was detected only in peripheral blood lymphocytes; 3) mixed populations of different subtypes of N-terminal and C-terminal sequences were observed in samples from one patient with nasopharyngeal carcinoma, one with T lymphoma and one with infectious mononucleosis sample; 4) intermediate variations between P-ala and V-val were observed in T-lymphoma, Hodgkin disease and infectious mononucleosis samples; and 5) in comparison with the major sequences identified in healthy carriers, the EBNA-1 sequences in peripheral lymphocytes from nasopharyngeal carcinoma were mixed types in 4 of 5 patients, implying increasing frequency of V-val might correlate with the progression of nasopharyngeal carcinoma.