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EB病毒DNA聚合酶基因的表达与纯化及其在鼻咽癌病人诊断中的应用 总被引:2,自引:0,他引:2
以Epstein-Barr病毒(EBV)DNA聚合酶为抗原,建立了简便、快速、敏感和特异的鼻咽癌诊断方法.构建原核表达载体pRSET-DNA聚合酶及其亚克隆pRSET-A1和pRSET-A2,在BL21(DE3)中表达的产物,经Western-blot检测其抗原性并用于检测鼻咽癌(nasopharyngeal carcinoma,NPC)病人血清中的抗体.在NPC病人血清中存在抗EB病毒DNA聚合酶的IgG抗体,并证明DNA聚合酶的抗原性主要集中于后2/3片段(A2)上.Western-bolt检测A2/IgG抗体的敏感性和特异性分别为80%和100%.对46份NPC病人血清和46份非NPC头颈癌症病人血清,用ELISA检测A2/IgA,敏感度为89%,特异度为93%.初步建立了ELISA检测NPC病人血清中A2/IgG抗体的方法,获得较高的敏感性和特异性. 相似文献
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应用测定Epstein-Barr病毒(EB病毒)IgA/EA抗体的改进方法(见病毒学报,2:372,1986).将待检血清用与抗人IgG血清或葡萄球菌菌体蛋白(SPA)吸附,除去了竞争性IgG类抗体,增加了血清中IgA抗体与抗原相结合的机率,从而提高了免疫酶法的敏感性和鼻咽癌的检出率。用此法我们检查了来本实验室检查EB病毒相关抗体的2045人,大部份是进行体格检查、无自觉症状的健康人,其查出IgA/EA抗体阳性者42人,进一步做病理检查证实27人患鼻咽癌,其中属临床Ⅰ期者10人,Ⅱ期10人,Ⅲ期6人, 相似文献
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已有大量研究工作证实,Epstein-Barr(EB)病毒与鼻咽癌的关系十分密切。因而,EB病毒在鼻咽癌病因学中的作用引起了人们的广泛重视。在EB病毒与宿主细胞相互关系的研究中,抗原表达与细胞隐性感染和细胞转化的相互关系,是目前所确认的关键课题之一。近年来的研究已经表明,EB病毒感染宿主细胞后,有多种EB病毒的特异性抗原产生,包括早期抗原(EA)、壳抗原(VCA)、膜抗原(MA)、核抗原(EBNA)、淋巴细胞确定的膜抗原(Lymphocyte-defined membrane antigen,LYDMA)和EB病毒潜 相似文献
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分析EB病毒核抗原1-IgA抗体(EBNA1-IgA)的不同状态及变化趋势,并探讨人群的鼻咽癌发病风险。检测中山市小榄镇2009~2010年入组的16 695位30~59岁参加鼻咽癌筛查人群血清,按首次筛查和随访中EBNA1/IgA抗体状态将筛查人群进行分组,对照组为未参加筛查的两个镇区同年龄组人群,利用中山市肿瘤登记系统、死因登记系统随访至2014年12月31日,分析各组人群的鼻咽癌发病风险。与对照组相比,基线抗体阴性人群的发病风险比为0.46(95%CI 0.25~0.86),基线抗体阳性人群的发病风险比为31.1(95%CI 21.0~46.1);复查人群中上升组、持续阳性组的发病风险比分别为82.4(95%CI 36.1~188.2),26.4(95%CI 12.3~52.5),而下降组和波动组中未见病例。EBNA1/IgA基线阳性人群在5年中有很高的发病风险,复查人群中上升组和持续阳性组也有很高的发病风险,下降组和波动组发病风险较低。 相似文献
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用萄葡球菌菌体A蛋白(SPA)预先处理被检血清,以去除抗体IgG部份的竞争性结合,提高了间接免疫荧光法检查鼻咽癌病人血清中EB病毒膜抗原IgA(IgA/MA)抗体的敏感性及特异性,检查48例鼻咽癌病人血清IgA/MA抗体,阳性率为100%,血清几何平均滴度为141;40例其它恶性肿瘤病人和46例正常人都检不出IgA/MA抗体,免疫荧光法测得IgA/MA抗体阴性的6例鼻咽癌病人血清,SPA吸收后呈阳性反应,此改进方法可用以追踪观察鼻咽癌病的病程及预后。 相似文献
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Background
The impact of variation of Epstein-Barr virus (EBV) antibody titers before the development of nasopharyngeal carcinoma (NPC) is still unclear. We analyzed the fluctuations of antibodies against EBV before histopathological diagnosis to assess the risk of NPC and aimed to provide a reliable basis for screening in high risk populations.Methods
This study was based on a population-based screening program in Sihui County in Guangdong Province of China. A total of 18,986 subjects were recruited in 1987 and 1992, respectively. Baseline and repeated serological tests were performed for IgA antibodies against EBV capsid antigen (VCA/IgA) and early antigen (EA/IgA). Follow-up until the end of 2007 was accomplished through linkage with population and health registers. Cox proportional hazards regression model was used to estimate the relative risk of NPC in association with EBV antibodies. Time-dependent receiver operating characteristic curve (ROC) analysis was used to further evaluate the predictive ability.Results
A total of 125 NPCs occurred during an average of 16.9 years of follow-up. Using baseline information alone or together with repeated measurements, serological levels of VCA/IgA and EA/IgA were significantly associated with increased risks for NPC, with a striking dose-response relationship and most prominent during the first 5 years of follow-up. Considering the fluctuant types of serological titers observed during the first three tests, relative risk was highest among participants with ascending titers of EBV VCA/IgA antibodies with an adjusted hazard ratio (HR) of 21.3 (95% confidence interval [CI] 7.1 to 64.1), and lowest for those with decreasing titers (HR = 1.5, 95% CI 0.2 to 11.4), during the first 5 years of follow-up. Time-dependent ROC analysis showed that VCA/IgA had better predictive performance for NPC incidence than EA/IgA.Conclusion
Our study documents that elevated EBV antibodies, particularly with ascending titers, are strongly associated with an increased risk for NPC. 相似文献12.
Clinical and immunological effect of intravesical interleukin-2 on superficial bladder cancer 总被引:2,自引:0,他引:2
TheBamHI Z EBV replication activator (ZEBRA) protein is involved in the switch from latency to productive cycle of Epstein-Barr virus. A recombinant ZEBRA protein was synthesized and assessed in enzymelinked immunosorbent assay (ELISA) for serum IgG response in nasopharyngeal carcinoma (NPC) patients. In 100 NPC serum samples that were positive for IgA to the EBV viral capsid antigen (VCA), 75% had IgG anti-ZEBRA antibodies. In contrast, only 3/83 (3.6%) serum samples from healthy donors and 2/50 (4%) from other cancers were positive for IgG to ZEBRA. Interestingly, in a selected group of 100 NPC sera negative for IgA to VCA, 25% contained IgG anti-ZEBRA antibodies. This suggests that the ELISA for IgG anti-ZEBRA may also identify earlier cases of NPC not detected by the conventional immunofluorescence test for IgA to VCA. 相似文献
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G De-Thé 《The Yale journal of biology and medicine》1981,54(1):33-39
Nasopharyngeal carcinoma (NPC), the major cancer killer in Cantonese Chinese, represents one of the three cancer research priorities in China. Apart from some fundamental research projects on the Epstein-Barr virus (EBV), their main efforts are epidemiologically and clinically oriented. Major surveys in GuangDong and GuangXi provinces showed that high EBV/IgA/VCA serum levels can detect NPC at an early stage of the disease. Characterization of "precancerous lesions" in the nasopharynx and among the EBV/IgA positive individuals is in progress in the Zangwu county of the GuangXi Autonomous Region. Preventive anti-viral intervention in these individuals is being discussed. Search for genetic and environmental chemical carcinogens is also carried out. This Chinese epidemiological approach complements well the fundamental Western studies on EBV and NPC. 相似文献
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近年来,常用未经丙酮固定的、由丁酸和巴豆油激活的B95-8细胞或P3HR-1细胞为靶细胞,检测人血清中EB病毒IgA/MA抗体以早期诊断鼻咽癌,效果良好。但由于B95-8细胞含有多种EB病毒抗原,不能用丙酮固定,需多次离心沉淀,在浮悬状态下检测,技术比较复杂。 相似文献
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Serological follow-up studies for up to 4 years on the levels of IgG antibody to EBV-determined nuclear antigen (EBNA) were carried out on 36 patients with nasopharyngeal carcinoma (NPC). The serum levels of IgA antibody specific to EBV capsid antigen (VCA) were also measured in some of the patients. The titers of EBNA antibody were measured by enzyme-linked immunosorbent assay (ELISA) and those of IgA antibody to VCA were measured by the indirect immunofluorescence method. The EBNA antibody titers in most sera from the patients before radiation therapy were found to be at least 4 times the mean values in the sera of healthy control adults. Within 2 to 8 months after completion of therapy by 4-MV liniac X-ray irradiation with total doses of 60 to 80 Gy, the titers of EBNA antibody in the sera of 6 patients had returned to normal levels, and low levels of EBNA antibody were maintained for a long time after therapy. These serological data were associated with a good clinical prognosis without recurrence or metastases. But in 6 patients, the patterns of change in the EBNA antibody levels were different: the levels remained high after therapy or first decreased to the normal level and then rose to at least 4 times this level. These 6 patients showed recurrence or metastases. The patterns of change in the EBNA antibody levels were well correlated with those of change in the levels of IgA antibody specific to VCA.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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R Glaser E C Strain K L Tarr J E Holliday R L Donnerberg J K Kiecolt-Glaser 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1985,179(3):352-355
Antibody titers to the Epstein-Barr virus (EBV), early antigen (EA) IgG, and virus capsid antigen (VCA) IgG and IgA, were measured in 44 geriatric subjects to determine if the depression in cellular immunity known to be associated with aging affects the expression of latent EBV. Similar assays were performed on plasma obtained from a young adult (medical student) population as a control group. We found that 89% of the geriatric samples were positive for EA IgG, and 83% of the plasma obtained from medical students were positive for EA IgG. One hundred percent of the geriatric subjects were positive for VCA IgG, and 87% of the medical students were positive for VCA IgG. Seven percent of the medical student blood samples were positive for VCA IgA; in contrast, 36% of the blood samples obtained from the geriatrics subjects were positive. Significant differences were also found in the geometric mean titers (GMT) of antibodies to EBV antigens; the GMT to EBV EA and VCA were significantly higher in the geriatric group. The data suggest that there may be some loss of control over latent EBV by the cellular immune response in geriatric individuals. 相似文献
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Z X Zhang W P Dong H S Chen Z G Xu Z H Zhang S M Liu E De Clercq 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1985,178(4):616-622
When persistently Epstein-Barr virus (EBV)-infected lymphoblastoid (B-95-8) cells were transplanted subcutaneously or intracerebrally to nude mice of either BALB/c or NIH background, tumors developed, and the tumor cells spontaneously expressed viral capsid antigen (VCA). This model was used to evaluate the in vivo anti-EBV activity of (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU), a highly potent and selective antiherpes agent, which was recently shown to inhibit several parameters of EBV infection in vitro. When administered intraperitoneally at 200 mg/kg/day for 4 weeks, or 500 mg/kg/day for 2 weeks, starting immediately after B-95-8 cell inoculation, BVDU effectively reduced tumor growth and VCA expression of either subcutaneously or intracerebrally inoculated B-95-8 cells. 相似文献
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山东临朐人群HLA等位基因多态性 与幽门螺杆菌感染关系的研究 总被引:4,自引:1,他引:3
为了研究山东临朐地区健康人白细胞抗原(HLA)Ⅰ类等位基因多态性与幽门螺杆菌(Hp)感染的关系,运用序列特异性引物聚合酶链反应(PCR SSP)的方法,检测Hp阳性人群(90例)和Hp阴性人群(49例)的HLA Ⅰ类和Ⅱ类等位基因。结果在HLA Ⅰ类(A、B、CW)的共68个等位基因多态中,发现在感染及非感染人群中存在4个显著性差异的位点;在HLA Ⅱ类(DRB1、DQB1和DRB3、DRB4、DRB5)的共22个等位基因多态中,没有发现显著性差异的位点。A*02等位基因频率,Hp阳性低于阴性人群(OR,0.56;95%CI,0.33~0.94;P,0.029);B*48等位基因频率,Hp阳性低于阴性人群(OR,0.15;95%CI,0.03~0.72;P,0.007);CW*08〖STBZ〗等位基因频率,Hp阳性低于Hp阴性人群(OR,0.32;95% CI,0.15~0.69;P,0.003);CW*15等位基因频率,Hp阳性高于Hp阴性人群(OR,5.11;95%CI,0.63~40.90;P,0.024)。结果表明HLA Ⅰ类等位基因的多态性可能与山东临朐地区Hp的易感性有关;HLA Ⅱ类等位基因的多态性可能与其无关。HLA Ⅰ类等位基因中,CW*15可能是Hp感染的易感基因;A*02、B*48和CW*08可能是保护性基因。Abstract: In order to analyze the relationship of HLA polymorphisms and the infection of H.pylori in the population of Linqu County in Shandong Province,polymerase chain reaction with sequence specific primers(PCR SSP) was used to determine the alleles of HLA typeⅠand Ⅱ in 90 Hp positive persons and 49 Hp negative controls.The results showed that among the 68 alleles of HLA typeⅠ,4 alleles were found significantly different between Hp positive and Hp negative population,while no significant difference was found among the 22 alleles of HLA typeⅡ.Hp positive persons had a lower allele frequency of A*02(OR=0.56,95% CI=0.33~0.94;P=0.029), B*48(OR=0.15,95% CI=0.03~0.72;P=0.007),CW*08(OR=0.32,95% CI=0.15~0.69;P=0.003)and a higher allele frequency of CW*15(OR=5.11,95% CI=0.63~40.90;P=0.024)compared with Hp negative controls.Our results indicated that the polymorphisms of HLA typeⅠis involved in the genetic susceptibility of Hp infection in Linqu County,while the polymorphisms of HLA typeⅡ may have no relationship with the genetic susceptibility of Hp infection.It was shown that among the alleles of HLA typeⅠ,CW*15might be a susceptible gene of Hp infection while A*02,B*48 and CW*08might be protective genes. 相似文献
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鼻咽癌病人和正常人群中EB病毒特异性T细胞对靶抗原的识别和应答 总被引:7,自引:0,他引:7
为探讨痘苗病毒表达的Eoskein-Barr病毒(EBV)核抗原1、4(EBNA1、4)和潜伏膜蛋白1、2(LMP1、2),在不同人群的特异性T细胞杀伤9CTL)中的作用,采集EBV阴性正常人、未经治疗的鼻咽癌(NPC)病人的EBV-IgA/VCA阳性者各10人的周围血淋巴单核细胞(PBMC),用EBV转化B淋巴细胞,建立类淋巴母细胞(LCL),用LCL刺激自休的T淋巴细胞作为效应细胞,以LCL感染重组痘苗病毒表达的EBNA1、4和LMP1、2为靶细胞,以^51Cr释放法检测EBV特异性CTL所识别的靶抗原。结果表明,EBV-LMP1、2可能既是EBV特异性T细胞的刺激抗原,又是其识别的靶抗原。将采集的30例试验者的各5 单克隆T细胞株分别检测HLA-Ⅰ型(A、B、C),按照不同型别寻找相对应的EBNA1、4和LMP1、2的不同合成肽,应用酶免疫吸附斑点法(Elispot)检测EBV特异性CD8^ 的CTL应答。结果显示:10例正常人中9人有特异的LMP2应答,4人有特异的EBNA4应答;10例未治疗的NPC病人中3人有特异的LMP2,2人有特异的EBNA1,3个有特异的EBNA4应答;10例未治疗的NPC病人中3人有特异的EBNA1,3人有特异的EBNA4应答,在10例EBV-IgA/VCAbj ntg k ,6人有特异的LMP2,5人有特异的EBNA4应答。所有的试验者均未发现LMP1的特异性应答。 相似文献