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1.
Background Epstein-Barr virus (EBV) associated malignancies with a Type Ⅱ latency gene expression pattern, such as Hodgkin‘s disease, and nasopharyngeal carcinoma (NPC) , frequently express the EBV antigen latent membrane protein 2A (LMP2A). We expected to establish a highly expressing LMP2A yeast cell strain and get the high quality LMP2A protein, which was used for detection, analysis and characterization of its antibodies in various patients‘ sera of EBV associated malignancies. Methods The plasmid pPICZαA-LMP2A containing the full length of LMP2A cDNA was constructed and transformed to Pichia pastoris GS115 to express LMP2A protein. After fermentation and purification, the LMP2A protein was used as an antigen to detect anti-LMP2A antibodies (Abs) in the sera of patients with EBVassociated malignancies in enzyme linked immunosorbent assay (ELISA) or Western-blot. Results LMP2A was expressed successfully with an expected molecular weight of approximately 54 kD and Abs to LMP2A were strikingly specific to NPC. Two-thirds or more sera from NPC patients were positive for antiLMP2A immunoglobulin G (IgG) Abs. The antibodies were absent from the sera of other EBV-associated diseases except a small fraction of the gastric carcinoma. Comparing anti-viral capsid Ags (VCA) IgA and LMP2A IgA titers in the sera from 76 NPC patients, only 55% were positive for anti-LMP2A IgA Abs while 70% were positive for anti-VCA IgA. However, we found that 3 sera negative for VCA IgA were positive for LMP2A IgA. Conclusion The results suggested the potential significance of LMP2A specific Abs for the diagnosis of EBVassociated malignancies, especially NPC.  相似文献   

2.
Objectives To investigate the immunophenotypes of primary nasopharyngeal non-Hodgkin lymphoma(NPL) and their relationship to Epstein-Barr virus (EBV) infection.Methods The clinical data and biopsies of 73 patients with NPL were collected in Guangzhou. In situ hybridization was performed to detect the EBV-encoded small non-polyadenylated nuclear RNAs(EBERs) on biopsy slides. Immunohistochemistry was used to classify the immunophenotypes of NPL and detect EBV antigen expression.Results Forty-four (60. 27%) of the 73 NPLs were of B cell lineage (CD79α^ /CD3^-/CD56^-)while the 29 others (39.73%) were of non-B cell lineage. Seventy-three NPLs could be classified into 3 major immunophenotypes: B cell (CD79α^ /CD3^-/CD56 ^-, 44 cases), peripheral T cell (CD79α^-/CD3^ /CD56^-, 22) and NK/T cell (CD79α^-/CD3^ /CD56^ , 7). The percentages of EBV infection differed among the 3 major immunophenotypes ( B cell : 11.36%, 5/44 ; peripheral T cell: 81.82%,18/22; NK/T cell: 100%, 7/7). Both CD56-positive and CD56-negative immunophenotypes could further be divided into 4 subtypes: CD8^-/CD4^-, CD8^ /CD4^- , CD8^-/CD4^ and CD8^ /CD4^ All the CD8^-/CD4^- NPLs with CD56-positivity (7) or CD56-negativity (2) were infected with EBV. The neoplastic cells of a nasopharyngeal Burkitt‘ s lymphoma expressed EBV nuclear antigen 1 (EBNA1)and EBV RNA (EBERs) only. In the other 29 EBV-infected NPLs, most of the lymphoma cells harboring EBV also expressed EBNA1 and EBERs; 21 of the 29 NPLs had a considerable number of neoplastic cells expressing latent membrane protein 1 ( LMP1 ) (21/29, 72.41% ) and 23 of 29 NPLs expressed latent membrane protein 2A (LMP2A) (23/29, 79. 31%). A few lymphoma cells in 17( 17/29, 58. 62% ), 23 (23/29, 79.31% ) and 22 NPLs (22/29, 75.86% ) expressed Zta ( Barn HI Z transactivator), viral capsid antigen (VCA) and membrane antigen (MA), respectively.Conclusions The prevalence ratio of the 3 immunophenotypes, namely, B cell, peripheral T cell and NK/T cell lymphoma, is about 6:3: 1. However, the EBV infection ratio is reversed, 1:8: 10. All the NK/T cell (CD56^ ) and peripheral immature T cell (CD3^ /CD8^-/CD4^-) NPLs were EBV-infected.Except for one Burkitt‘ s lymphoma, the EBV harbored in both B cell and non-B cell NPLs was mainly latent infection, type It, expressing EBNA1, LMP1 and LMP2A. However, the EBV found in a few lymphoma cells could become replicative, expressing lytic proteins.  相似文献   

3.
The expression of DNA ploidy, the cell cycle and Ki67 antigen in nasopharyngeal carcinoma (NPC) were studied and their relationship with the clinical biological behaviors and prognosis of NPC was evaluated. Biopsied specimens of NPC were made into cell suspension. By using cytometric double labeling Ki67 and DNA method, the expression of DNA ploidy, the cell cycle and Ki67 antigen were analyzed. The patients were followed up for about 3 years and the relationship between the above mentioned parameters and the clinical biological behavior and prognosis of NPC were evaluated. Of the 62 cases of NPC, the DNA aneuploid accounted for 29.03 %. The S phase cells accounted for 0 to 54 % in the cell cycle and the positive expression of Ki67 ranged from 0 to 52%. There were 40 cases of LPI (64.5%) including 15 negative cases and 22 cases of HPI (35.5%) respectively. The DNA anneploid content was positively related to the S phase cells. The patients having a low expression of Ki67 or DNA aneuploid in tumor cells were not sensitive to chemotherapy, liable to metastasis to distant organs and had a poor prognosis, while Ki67 showed no correlation with DNA ploidy and the cell cycle. It was suggested that DNA ploidy and Ki67 could be used as an independent and objective marker to evaluate the radiosensitivity and prognosis of NPC.  相似文献   

4.
5.
Objective To study the effects of dendritic cells(DC) transfected with recombinant vaccinia virus encoding Epstein-Barr virus(EBV) latent membrane protein 2A(LMP2A) gene,and to provide evidence for further investigation on the therapeutic uaccines against EBV-associated malignancies.Methods Mature DC were transfected with EVB-LMP2A recombinant vaccinia virus(rVV-LMP2A).Before and after the transfection,the expression of surface antigens on mature DC including CD1a,CD83,CD40,CD80,HLA-DR was measured by fluorescence activated cell sorter(FACS) and the function of DC to stimulate allogeneic T cells proliferation was measured by mixed leukocyte reactions(MLR).Results LMP2A protein was highly expressed (66.1%) in DC after the transfection of rVV-LMP2A.No significant changes in the primary surface antigens expression and in the MLR were detected during the transfection.Transfected DC still had strong potential in stimulating the proliferation of allogeneic T cells.Conclusion Recombinant vaccinia virus was an effective and non-perturbing vector to mediate the transfection of LMP2A into DC.The functions of mature DC were not affected significantly by the transfection of Vac-LMP2A.This study could provide evidence for the further immunotherapy of EBV-associated malignancies,e.g.nasopharyngeal carcinoma(NPC).  相似文献   

6.
Background Epstein-Barr virus (EBV) is a herpesvirus commonly associated with several malignant diseases including nasopharyngeal carcinoma (NPC), which is a common cancer in Southeastern Asia. Previous studies showed that plasma levels of EBV-DNA might be a sensitive and reliable biomarker for the diagnosis, staging and evaluating of therapy for NPC. There are a few analyses of the levels of EBV-latent membrane protein 2 (LMP2)-specific cytotoxic T-lymphocytes (CTLs) in patients with NPC. This study was conducted to investigate the levels of EBV-LMP2-specific CTLs, EBV-DNA load and the level of CD4^+CD25^+T cells in such patients.
Methods From February 2006 to April 2006, 62 patients with NPC, 40 healthy virus carriers positive for EBV viral capsid antigen (EBV-IgA-VCA) and 40 controls were enrolled in the study. We used a highly sensitive ELISPOT assay, real-time polymerase chain reaction (PCR) and flow cytometry to measure the EBV-LMP2-specific CTL response, the EBV DNA load and the level of CD4^+CD25^+T cells, respectively.
Results The EBV-LMP2-specific CTL responses of the samples from the control, healthy virus carriers and patients with NPC were significantly different from the LMP2 epitopes, with the control and healthy virus carrier samples displaying a stronger response in three cases. There were significant differences in EBV DNA load in serum between NPC and the healthy groups; patients with NPC at stages Ⅲ or Ⅳ had significantly higher viral loads compared with those at stages Ⅰ or Ⅱ. A significantly higher percentage of CD4^+CD25^+ T lymphocytes were detected in the patients, compared with healthy virus carriers and healthy controls. Moreover, patients with advanced stages of NPC (Ⅲ and Ⅳ) had significantly higher percentages than the patients with early stages (Ⅰ and Ⅱ). Conclusions Patients with NPC are frequently unable to establish or maintain sufficient immunosurveillance to control proliferating B cells harboring EBV and to destroy the tumor cells that express immunodominant LMP2 proteins.
Controlling the activity of CD4^+CD25^+T cells and elevating CD8^+ cells specific for LMP2 epitopes could be an effective immunotherapy for patients with NPC.  相似文献   

7.
8.
The effect of vascular endothelial growth factor (VEGF) overexpression on matrix met-alloproteinase-2 (MMP-2) in nasopharyngeal carcinoma (NPC) cells in vitro and the possible mechanism involved were investigated, and the correlation between the expression of VEGF and MMP-2 in NPC evaluated. The NPC cells were transfected with PAd-trackVEGF165 plasmid. The expression levels of VEGF and MMP-2 mRNA and protein in NPC cells were detected by semi-quantitative RT-PCR and Western blot respectively. It was found that the expression of VEGF and MMP-2 mRNA and protein was significantly increased in NPC cells after transfection of VEGF165. It was concluded that the expression of VEGF was correlated to the in vitro invasion of NPC cells, and the induction of MMP-2 by VEGF was a key process of NPC cell invasion.  相似文献   

9.
Objective To compare the detection rates of Epstein-Barr virus (EBV) DNA in the serum/plasma between apparently healthy adults (AHAs) and nasopharyngeal carcinoma (NPC) patients in attempt to evaluate the efficiency of EBV DNA assay for serodiagnosis of NPC.Methods The plasma and serum were obtained from 58 AHAs and 66 untreated NPC patients. EBV DNA W-fragment was detected using nested ploymerase chain reaction (PCR). Immunoenzymatic assay for titration of IgA-VCA was also adopted. Results EBV DNA detection rate (84.85%) in the plasma/serum of 66 NPC patients was significantly higher than that (10.34%) in 58 AHAs. The sensitivity of plasma/serum EBV DNA assay (0.8485) was higher than that (0.8030) of titrating IgA-VCA (positive criterion≥1∶40) though the specificities of these two tests were the same (0.8966). The correct rate, predictive value of a positive test, and Odds ratio of dual positivity (0.8387, 0.9792 and 141.0, respectively) were higher than those of single positivity either to plasma/serum EBV assay (0.5242, 0.7333 and 1.1423, respectively) or to IgA-VCA≥1∶40 test (0.4839, 0.5385 and 1.0480, respectively). Conclusion The EBV DNA detection in the plasma/serum using nested PCR may be a useful indicator for serodiagnosis of NPC.  相似文献   

10.
Background Intestinal T-cell lymphoma (ITCL) is a heterogeneous lymphoid neoplastic group with variable clinical and pathological features. ITCL in oriental countries is different from enteropathy-type intestinal T-cell lymphoma (ETCL) in relation to celiac disease and Epstein-Barr virus (EBV). The objective of this study was to investigate the clinicopathological features, immunophenotype, expression of cytotoxic molecule (TIA-1), T-cell receptor (TCR)-γ gene rearrangement, and Epstein-Barr virus (EBV) latent infection in primary ITCL without celiac disease in Chinese.Methods The clinical data of 42 patients were analyzed, and the patients were followed up. Compared with human reactive lymphoid tissues, in situ hybridization for EBER1/2, polymerase chain reaction for TCR-γ gene rearrangement, and immunohistochemical staining for immunophenotypes, TIA-1 and EBV latent membrane proteins (LMP-1) were investigated. Survival curves of different clinicopathological features, immuno-phenotypes, expression of LMP1, TCR-γ gene rearrangement and therapy were analyzed.Results Three fourths of the patients suffered from ITCL in China were men with a peak age incidence in the 4th decade. Common presenting features included fever and hemotochezia. The prognosis was poor with a median survival of 3.0 months. The lesions were mostly localized in the ileocecum and colon. About 38/42 (90.5%) patients demonstrated pleomorphic medium-sized on large cells. Histological features of celiac disease were rarely seen. All 42 patients with ITCL revealed CD45RO positive. Neoplastic cells partially expressed T-cell differentiated antigens (CD3ε, CD4, CD8) and NK cell associated antigen (CD56). The positive frequency of CD3ε, CD4, CD8 and CD56 was 28/42 (66.7%), 7/42 (16.7%), 10/42 (23.8%) and 12/42 (28.6%) respectively. Thirty-nine cells (92.9%) expressed TIA-1, but none expressed CD20 and CD68. More than half of the patients (64.3%, 64.3% and 59.5%) revealed TCR-γ gene rearrangement by three different TCR-γ primers respectively. EBER1/2 was detected in 41 (97.6%) of the 42 patients. The expression frequency of LMP-1 was 38.1% (16/42).Conclusions Primary ITCL without celiac disease in Chinese is a special highly EBV-associated clinicopathological entity. There are few similarities in patients with celiac disease in western countries. A small proportion of primary ITCLs in Chinese and extranodal NK/T-cell lymphoma of nasal type belong to the same spectrum.  相似文献   

11.
目的:探讨凋亡抑制基因Survivin蛋白和EB病毒潜伏膜蛋白(LMP1)在鼻咽癌组织中的表达情况以及它们之间的关系.方法:应用免疫组织化学技术S-P法对68例鼻咽癌组织标本进行凋亡抑制基因Survivin蛋白与EB-LMP1的表达检测.结果:鼻咽癌组织Survivin蛋白产物的阳性率为82.4%(56/68),LMP1阳性率为67.6%(46/68);56例Sur-vivin蛋白表达的阳性鼻咽癌标本中,有42例LMP1阳性,共同阳性率75.0%(42/56),12例Survivin蛋白表达的阴性鼻咽癌标本中,9例LMP1阴性,共同阴性率66.7%(9/12),两者之间高度相关(P〈0.01).结论:鼻咽癌组织中凋亡抑制基因Survivin蛋白的表达与LMP1的表达密切相关.  相似文献   

12.
应用抗EB病毒膜抗原(EBV/MA)的单克隆抗体检测51例病理确诊的鼻咽癌(NPC)活检标本的冰冻切片,其中49例有EBV/MA的表达。经细胞形态学分析首次发现鼻咽癌癌细胞,增生上皮细胞以及正常鼻咽部上皮细胞都有EBV/MA。以Southern杂交法检查20例鼻咽癌活检组织中EB病毒核酸(EBV/DNA),结果证实鼻咽癌组织中不仅有EB病毒核酸的重复序列W片段,还有与细胞转化有关的K片段的序列存在。进一步表明EB病毒在鼻咽癌的发生中并非“过客”,很可能是其病原因素之一。  相似文献   

13.
目的:探讨P53和P21蛋白在鼻咽癌(NPC)、癌旁组织和鼻咽慢性炎症中表达的差异,及其与肿瘤生物学行为的关系。材料与方法:首程放疗前的NPC病人100例,癌旁组织和鼻咽慢性炎症对照组各20例,用SP法进行P53和P21蛋白免疫组化染色,P53阳性细胞的标准为:癌巢内肿瘤细胞核有棕色沉着的细胞;P21阳性细胞的标准为:癌巢内肿瘤细胞浆有棕色沉着的细胞。结果:在鼻咽癌、癌旁组织和鼻咽慢性炎症组织中,P53蛋白的阳性率分别为:78.0%、35.0%和0.0%,P=0.001;P21蛋白的阳性率分别为:83.0%、75.0%和65.0%,P=0.170。在鼻咽癌原发灶早期(T1+2)和晚期(T3+4),颈结阴性和阳性及临床早期(Ⅰ Ⅱ)和临床晚期(Ⅲ Ⅳ)各组中,P53蛋白和P21蛋白的阳性率均无差异(P>0.05)。结论:P53基因的突变与鼻咽癌的发生有一定的关系。P53蛋白可能是鼻咽癌癌病变的指标;P21的突变可能与鼻咽癌的发生无明显的关系,P53蛋白和P21蛋白的阳性率与鼻咽癌原发灶的早晚,颈结转移,和临床分期无关。  相似文献   

14.
目的:对重组克隆表达抗原血清学分析(SEREX)法筛选出的鼻咽癌(NPC)系列肿瘤抗原进行特异性检测,判断新检测出的NPC抗原的相应抗体在NPC病人血清及正常人血清中的阳性率,为NPC抗原对NPC的免疫诊断、免疫治疗积累基础资料。方法:运用噬菌体转染-蛋白印迹、免疫组织化学技术,检测3种NPC肿瘤抗原(GX-NPC-1,GX-NPC-2,GX-NPC-4)在20个NPC病人及15个正常人血清中的阳性表达率。结果:3种NPC肿瘤抗原在NPC病人血清中的阳性表达率分别为55%(11/20)、55%(11/20)、50%(10/20);正常人各为33.3%(5/15)、26.7%(4/15)、6.7%(1/15),经统计学分析,前两种肿瘤抗原在NPC及正常人血清中的表达差异无统计学意义,而GX-NPC-4在两种血清中的表达差异有统计学意义(P<0.01)。结论:3种NPC肿瘤抗原中GX-NPC-4抗原在NPC患者的血清中的表达率明显增高,可能为NPC特异性抗原,值得进一步研究。  相似文献   

15.
鼻咽癌组织中EBER-1表达及其临床病理学意义   总被引:3,自引:0,他引:3  
目的:探讨爱泼斯坦-巴尔病毒(EBV)与鼻咽癌发生、发展的关系及其临床病理学意义。方法:应用免疫组化技术检测46例鼻咽癌、28例鼻咽癌癌旁组织、36例慢性鼻咽炎石蜡切片组织中爱泼斯坦-巴尔病毒受体(EBER-1)的表达。结果:鼻咽癌组织中EBER-1表达率为74.O%,且在转移灶中不丢失,癌旁组织、慢性鼻咽炎组织中EBER-1表达率分别为32.1%、16.7%。结论:检测EBER-1的表达对鼻咽癌诊断与鉴别诊断具有重要价值。慢性鼻咽炎组织、癌旁组织中:EBER-1阳性者提示为高危癌前病变,应密切随访监测,可能有利于早期发现鼻咽癌。  相似文献   

16.
 【目的】 检测EB病毒潜伏膜蛋白1 (LMP1) 远端启动子L1-TR在鼻咽癌(NPC)组织中的突变情况并探讨突变对启动子活性的影响。【方法】 用聚合酶链反应(PCR)方法分别扩增出B95.8细胞和新鲜鼻咽癌组织细胞中EB病毒(EBV)的L1-TR片段,并测序比较其核苷酸序列的差异。构建含原型和突变型L1-TR启动子的荧光素酶报告质粒并分别转染HaCat细胞、B95.8细胞和C666-1细胞,比较两种启动子的活性。【结果】 和B95.8原型比较,鼻咽癌组织中的EB病毒L1-TR启动子区域有多处缺失。插入和点突变,且启动子活性明显降低(P < 0.05)。【结论】 鼻咽癌组织中LMP1启动子 L1-TR的活性与B95.8原型相比明显降低,这在EB病毒的免疫逃避机制中可能有一定的意义  相似文献   

17.
EB病毒感染与宿主细胞恶性转化的关系   总被引:2,自引:0,他引:2  
罗春艳  甘润良 《医学综述》2009,15(11):1613-1617
EB病毒(EBV)是一种与多种肿瘤发生密切相关的DNA肿瘤病毒。EBV基因表达产物有潜伏膜蛋白(LMP1、LMP2A、LMP2B),6种核抗原(EBNA1、EBNA2、EBNA3A、EBNA3B、EBNA3C、EBNA-LP),以及EBER1、EBER2和TP等。EBV通过影响和调控宿主细胞基因的表达,干扰细胞信号转导系统,使宿主细胞产生细胞因子表达失衡。EBV能上调IRF-4、IL-6、Bcl-6、Bcl-2、A20等细胞生长因子、抗凋亡因子,还能抑制p53和PTPPK等肿瘤抑制基因,促使宿主细胞转化为肿瘤细胞。  相似文献   

18.
鼻咽癌患者血清中EB病毒四种特异性IgG抗体的测定   总被引:2,自引:0,他引:2  
用间接免疫荧光法检测了24例鼻咽癌患者与119例正常人血清中EB病毒4种特异性IgG抗体,结果表明,正常人血清中EB病毒衣壳抗原(WCA)的抗体效价为1:160以下,而鼻咽癌患者血清抗体效价为l:160~1:5120,抗体阳性率均为100%;正常人血清中EB病毒核抗原(EBNA)的抗体效价为1:80以下,而鼻咽患者血清抗体价为1:80~1:2560,阳性率均为100%;正常人血清中EB病毒早期抗原(EA)的抗体效价为l:10,阳性率只有13.3%,而鼻咽癌患者血清抗体效价为l:20~1:1280,阳性率为100%;正常人血清中EB病毒抗原(MA)的抗体效价为1:10~1:40,阳性率为71%,而鼻咽癌患者血清抗体效价为1:10~1:320,阳性率为100%.以上4种抗体中.VCA、EBNA和EA抗体的效价和分布在正常人与鼻咽癌患者之间有明显差异(P<姜勇男,男,31岁,讲师.延吉市局子街21号(133000)0.0l),EA抗体的阳性率也比正常人明显升高(P<0.0l),MA抗体的效价和阳性率与正常人相比无显著差异(P>0.05)。  相似文献   

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