Prospective observation: Clinical utility of plasma Epstein–Barr virus DNA load in EBV-associated gastric carcinoma patients

Epstein–Barr virus (EBV)-associated gastric carcinomas (EBVaGCs) may account for 8–9% of all gastric cancer (GC) patients. All previous reports on EBVaGC were retrospective. Prospective study is warranted to evaluate the exact role of EBV status in predicting the prognosis of GC. It is of special interest to figure out whether dynamic detection of plasma EBV-DNA load could be a feasible biomarker for the monitor of EBVaGC. From October 2014 to September 2017, we consecutively collected GC patients (n = 2,760) from Sun Yat-sen University Cancer Center for EBER examination. We detected EBV-DNA load in plasma and tissue samples of EBVaGC patients at baseline. Subsequently, plasma EBV-DNA load was dynamically monitored in EBVaGC patients. The overall prevalence of EBVaGC is 5.1% (140/2,760). The incidence rate of EBVaGC decreased with advanced AJCC 7th TNM stage (p < 0.001), with the corresponding percentages of 9.3, 9.9, 6.7 and 1.4% for Stage I, II, III and IV patients. EBVaGC patients were predominately young males with better histologic differentiation and earlier TNM stage than EBV-negative GC (EBVnGC) patients. EBVaGC patients were confirmed to had a favorable 3-year survival rate (EBVaGC vs. EBVnGC: 76.8% vs. 58.2%, p = 0.0001). Though only 52.1% (73/140) EBVaGC patients gained detectable EBV-DNA and 43.6% (61/140) reached a positive cutoff of 100 copies/ml, we found the plasma EBV-DNA load in EBVaGC decreased when patients got response, while it increased when disease progressed. Our results suggested that plasma EBV-DNA is a good marker in predicting recurrence and chemotherapy response for EBVaGC patients.     

Clinical significance of variations in levels of Epstein-Barr Virus (EBV) antigen and adaptive immune response during chronic active EBV infection in children

Abstract

Pediatric patients were recruited to analyze differences in Epstein-Barr virus (EBV) copy numbers and adaptive immune reactions in children with chronic active vs acute EBV infection (CAEBVI vs AEBVI), as well as to examine the relationship between these parameters and the pathogenesis of CAEBVI. Fluorescent qPCR was used to assess EBV-DNA levels, while ELISA, antibody affinity, flow cytometry, and heterophil agglutination (HA) assays were used to evaluate patient EBV-adaptive humoral and cellular immunity. Lastly, ELISPOT was employed to assess interferon (IFN)-γ secretory functions of EBV-specific cytotoxic T-lymphocytes (CTL) as a marker of subject EBV-specific adaptive cellular immunity. The results indicated that, compared with AEBVI patients or normal children, there was a dramatic elevation in viral copy levels, viral capsid antigen (VCA)-IgA, early antigen (EA)-IgA, and EA-IgG, but a lack of EBV nuclear antigen (EBNA)-IgG and a negative HA in CAEBVI patients (p?<?0.01). These subjects also had decreased CD4⁺, CD8⁺ (naïve), CD8⁺CD38⁺, and effective memory T-lymphocyte levels compared with AEBVI patients (p?<?0.01), and decreased EBV-specific CTL function compared with normal children (p?<?0.01). These results suggest that there is a disturbance in EBV antigen availability and in both the adaptive humoral and cellular immune responses in patients with CAEBVI, and that these outcomes may be associated with the chronic active re-infection process itself associated with CAEBVI.     

血浆EB病毒DNA 定量测定对鼻咽癌的诊断价值

目的分析血浆EB病毒（Epstein-Barr virus,EBV）DNA定量测定对鼻咽癌的诊断价值。方法采用荧光定量聚合酶链反应（fluorescence quantitative polymerase chain reaction,FQ-PCR）技术,测定35例经病理确诊的鼻咽癌患者血浆EB病毒DNA含量,并与50例健康对照者比较。结果鼻咽癌患者血浆EBV-DNA阳性率和复制量均高于健康对照者。两组阳性率和中位浓度的比较均有统计学差异。结论血浆EBV-DNA定量分析不能作为临床上确诊鼻咽癌的方法,但对鼻咽癌的筛查有实用价值,值得进一步研究。     

鼻咽癌患者血清中VCA-IgA、EA-IgA、EBV-DNA评价的比较

目的比较分析外周血EB病毒衣壳抗原抗体(VCA-IgA)、早期抗原抗体(EA-IgA)和EB病毒DNA(EBV-DNA)在鼻咽癌(NPC)诊断中的意义。方法对123例鼻咽癌患者、50例鼻炎患者、40例健康成人血清用酶联免疫吸附反应(ELISA)法检测VCA-IgA、EA-IgA抗体,荧光定量PCR法检测EBV-DNA,分析比较这3种检验方法的灵敏度与特异性。结果VCA-IgA、EA-IgA、EBV-DNA的灵敏度和特异性分别是35.77%和97.78%,71.54%和95.56%,26.83%和97.78%。EA-IgA的灵敏度最高,其次是VCA-IgA。VCA-IgA和EBV-DNA的特异性均稍高于EA-IgA。结论VCA-IgA和EA-IgA抗体测定与EBV-DNA联合检测可作为筛查鼻咽癌患者、观察鼻咽癌治疗效果和预测预后的辅助手段。     

血浆EBV-DNA与VCA-IgA联合检测对鼻咽癌诊断的意义

目的探讨血浆EBV-DNA水平与VCA-IgA联合检测对鼻咽癌诊断的临床意义。方法收集鼻咽癌患者88例,健康体检者45例作为对照组。采用荧光定量PCR方法检测EBV-DNA水平,采用酶联免疫吸附法检测VCA-IgA抗体滴度。结果鼻咽癌患者中,EBV-DNA阳性率81.82%,VCA-IgA阳性率89.77%,健康体检者中,EBV-DNA阳性率6.67%,VCA-IgA阳性率31.11%,两组阳性率有显著差异(P<0.01);早期(I、Ⅱ期)鼻咽癌患者EBV-DNA水平及VCA-IgA滴度与晚期(III、IV期)鼻咽癌患者EBV-DNA水平及VCA-IgA滴度有显著差异(P<0.05)。结论血浆EBV-DNA与VCA-IgA联合检测对鼻咽癌的诊断有极高价值,可作为监测病情的一种有效手段。     

EB病毒在复发鼻咽癌患者中的研究进展

目的 了解EB病毒（爱泼斯坦 - 巴尔病毒,Epstein - barr virus,EB病毒）与复发鼻咽癌的关系及目前研究进展。方法 查阅相关文献,就EB病毒与初治、复发鼻咽癌的关系,EB病毒在复发鼻咽癌中的致病机制进行综述。结果 EB病毒潜伏感染是鼻咽癌发病的最主要危险因素,95%以上的初治鼻咽癌与EB病毒感染相关。复发鼻咽癌患者中,在鼻咽组织及外周血中可检测到EBV - DNA（Epstein - barr virus - deoxyribonucleic acid,EBV - DNA）,鼻咽组织EBV - DNA阳性率甚至可高达100%,鼻咽癌复发时血浆EBV - DNA 平均浓度可用于预测肿瘤转移和再次复发。然而,EB病毒在复发鼻咽癌中的致病机制尚未明确,其分子通路需要进一步研究。结论 EBV - DNA作为复发鼻咽癌诊断的理想生物标志物,能实现早预测、早诊断、早治疗复发鼻咽癌,对EB病毒在复发鼻咽癌癌中作用机制的研究,具有广阔的发展前景。     

鼻咽癌患者放疗前后外周血EBV-DNA和细胞免疫水平的研究

摘要：目的比较鼻咽癌（NPC）患者放疗前后抗EBV特异性CTL细胞免疫功能、EBV-DNA、T细胞亚群及CD4+CD25+Tr淋巴细胞水平,为临床选择免疫治疗方案及预测预后提供理论依据及客观指标。方法收集标本105份（NPC患者放疗前35份、NPC患者放疗6月后40份和健康人30份）,ELISPOT检测外周血LMP2特异性CTL;荧光定量PCR检测EBV-DNA;流式细胞术检测T细胞亚群及CD4+CD25+Tr淋巴细胞,统计分析各指标变化的特点及相互关系。结果抗EBV-LMP2特异性细胞免疫反应最大频率见于健康对照组,NPC放疗前患者中则仅少数呈阳性反应,NPC放疗后组反应率介于NPC放疗前组和健康对照组之间;NPC放疗前组血清EBV-DNA的阳性率均高于其他两组,对EBV-LMP2特异性细胞免疫反应的阳性率与血清中EBV-DNA病毒载量的阳性率呈负相关;放疗前NPC患者外周血CD3+、CD4+T细胞百分率及CD4+/CD8+比值降低, CD8+T细胞百分率升高,CD4+CD25+Tr细胞水平升高,放疗后情况有所缓解。结论EBV-LMP2特异性CTL反应在NPC放疗前患者最低,EBV-LMP2特异性CTL反应阳性率与血清中EBV-DNA载量相关,增强CTL应答可改善EBV相关肿瘤的治疗效果,EBV-DNA载量水平一定程度上可反映NPC患者的肿瘤负荷。NPC患者外周血CD3+、CD4+T细胞降低,而CD8+T细胞升高,CD4+CD25+Tr细胞比例升高,监测患者T细胞亚群有助于对NPC患者的治疗及预后的判断。     

鼻咽癌患者血浆EBV-DNA检测的临床价值

目的 EB病毒与鼻咽癌的发生发展明确相关,多个研究显示血浆EBV-DNA对于鼻咽癌的诊断分期及预后判断有重要意义,因此对我院患者进行了血浆EBV-DNA的检测并分析其临床价值。方法 2013—2016年连续性鼻咽癌患者471例,分析其治疗前EBV-DNA水平与分期、肿瘤负荷的相关性,并对治疗前、治疗末EBV-DNA进行生存相关分析。结果 患者治疗前血浆EBV-DNA中位数137 copies/ml (0~494000),与T分期、N分期、M分期、总的临床分期、肿瘤负荷均有明显统计学相关性。生存分析显示治疗前血浆EBV-DNA拷贝数≤1300组比>1300组的患者有更好OS (P=0.007)、PFS (P=0.011)、DMFS (P=0.003)。治疗末血浆EBV-DNA不可检测到的患者有更好OS (P=0.016)、PFS (P=0.000)、DMFS (P=0.000)。Cox多因素分析T分期、治疗末是否可检测到EBV-DNA为OS (P=0.030、0.012)的预后因素,N分期(P=0.037、0.017)、放疗末是否可检测到EBV-DNA (P=0.006、0.001)为PFS及DMFS的预后因素。结论 鼻咽癌患者治疗前血浆EBV-DNA水平与分期及肿瘤负荷有明显相关性,治疗前EBV-DNA水平更高的患者可能治疗后的预后更差。治疗末血浆EBV-DNA是否可检测到对于OS、PFS、DMFS有较好的预测价值。     

EB病毒DNA在鼻咽病变中的表现

本文通过原位杂交技术检测鼻咽活检组织不同病理改变时EBV—DNA的表现,以探讨EB病毒在鼻咽癌发生过程中的可能作用。结果显示:(1)本实验所有鼻咽低分化癌癌组织均出现EBV—DNA片段,且绝大部分病例的阳性细胞数及细胞阳性强度均在中等以上;(2)无论是鼻咽癌或慢性鼻咽炎病例,所见到的中-重度异型改变上皮或淋巴组织,均有数量不等的细胞存在阳性强度不同的EBV-DNA片段,粘膜下小涎腺内亦常见到一定数量的EBV-DNA片段。结果提示:(1)EB病毒与鼻咽低分化癌关系密切。EBV作为致鼻咽癌病因多因素中的一员是有可能的;(2)患者血清中EB病毒相关抗体,尤其是IgA抗体阳性率和滴度的升高,可能与鼻咽局部EBV感染及/或EBV激活有关;(3)在异型改变上皮中有中等或以上阳性强度EBV-DNA片段的检出,将有可能作为推测鼻咽癌癌前病变的有效指标。     

鼻咽癌患者EBV、HPV共感染状况真实世界初步研究

目的:初步分析四川南充地区鼻咽癌患者EB病毒（EBV）与人乳头瘤病毒（HPV）共感染状况及其临床意义。方法:收集经鼻咽镜活检证实的初诊鼻咽癌患者的石蜡包埋组织,采用分子杂交技术以及PCR荧光定量行EBV核酸定量（EBV-DNA）检测及HPV分型检测,分析EBV、HPV共感染状况与人口学特征、分期、治疗相关副反应、近期疗效等指标的相关性。结果:鼻咽癌患者总体EBV感染率为81.7%,HPV感染率为40%,28.3%的鼻咽癌患者存在HPV和EBV共感染;中晚期患者EBV感染高于HPV感染,EBV单感染患者CR率为29.4%,HPV感染（HPV单感染+EBV/HPV共感染）患者CR率为63.6%,差异接近有统计学差异（P=0.074）,四种感染状态在性别、年龄、吸烟状况、T分期等方面无差异。结论:鼻咽癌EBV感染率高于HPV,中晚期患者EBV感染率更高,<60岁患者HPV感染更常见;鼻咽癌患者存在一定比例EBV/HPV共感染状态;鼻咽癌EBV感染背景下,合并HPV感染可能是预后良好的一个分子标记物。