Benjamin E Warner,Japan Patel,Renwei Wang,Jennifer Adams-Haduch,Yu-Tang Gao,Woon-Puay Koh,Ka Wo Wong,Alan K S Chiang,Jian-Min Yuan,Kathy H Y Shair
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Denatured lysates were evaluated by a refined IgA and IgG immunoblot assay in a case-control study using pre-diagnostic NPC sera from two independent cohorts in Singapore and Shanghai, P.R. China.\r\n\r\nRESULTS\r\nAt 95% sensitivity, 487V yielded a 94.9% specificity compared to 86.1% for 487A. EBNA1 deleted for the conserved glycine-alanine repeats (GAr) reduced false positives by 22.8%. NPC sera reacted more strongly to the C-terminus than healthy controls, but the C-terminal construct (a.a. 390-641) showed lower specificity (84.8%) than the EBNA1 GAr deleted construct (92.4%) at 95% sensitivity.\r\n\r\nCONCLUSION\r\nAlthough EBNA1 IgA was present in healthy sera, most epitopes localized to the immunodominant GAr. We conclude that a refined EBNA1 antigen deleted for the GAr but with residues consistently detected in Southeast Asian NPC tumors is optimal for risk prediction with an extended sojourn time of 7.5 years. Furthermore, distinct EBNA1 serologic profiles enhanced the utility of the EBNA1 IgA assay for risk stratification. 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引用次数: 0
摘要
目的选择性爱泼斯坦-巴氏病毒(EBV)蛋白抗体可诊断早期鼻咽癌(NPC)。我们以前的研究表明,针对 EBV 核抗原 1 (EBNA1) 的 IgA 可以预测高危和中危人群在诊断前 4 年发生的鼻咽癌。设计开发了哺乳动物表达的构建体来代表 EBNA1 变体 487V 和 487A,这两种变体的 N 端和 C 端相差≥15 个氨基酸。在一项病例对照研究中,使用来自新加坡和中国上海两个独立队列的诊断前鼻咽癌血清,通过精制的 IgA 和 IgG 免疫印迹分析法对变性裂解液进行了评估。结果在 95% 的灵敏度下,487V 的特异性为 94.9%,而 487A 为 86.1%。EBNA1 删除了保守的甘氨酸-丙氨酸重复序列 (GAr),使假阳性率降低了 22.8%。与健康对照组相比,鼻咽癌血清对 C 端的反应更强,但在 95% 的灵敏度下,C 端构建体(a.a. 390-641)的特异性(84.8%)低于 EBNA1 GAr 删除构建体(92.4%)。我们得出的结论是,删去了 GAr 但在东南亚鼻咽癌肿瘤中持续检测到残基的改良 EBNA1 抗原是风险预测的最佳抗原,其存活时间可延长至 7.5 年。此外,不同的 EBNA1 血清学特征增强了 EBNA1 IgA 检测对风险分层的作用。这说明了血清学相关的 EBNA1 序列对于鼻咽癌风险预测和早期检测的重要性。
The Epstein-Barr virus nuclear antigen 1 variant associated with nasopharyngeal carcinoma defines the sequence criteria for serologic risk prediction.
PURPOSE
Antibodies to select Epstein-Barr virus (EBV) proteins can diagnose early-stage nasopharyngeal carcinoma (NPC). We have previously shown that IgA against EBV nuclear antigen 1 (EBNA1) can predict incident NPC in high- and intermediate-risk cohorts 4 years pre-diagnosis. Here, we tested EBNA1 variants, with mutants, to define the sequence requirements for an NPC risk assay.
DESIGN
Mammalian-expressed constructs were developed to represent EBNA1 variants 487V and 487A which can differ by ≥15 amino acids in the N- and C-termini. Denatured lysates were evaluated by a refined IgA and IgG immunoblot assay in a case-control study using pre-diagnostic NPC sera from two independent cohorts in Singapore and Shanghai, P.R. China.
RESULTS
At 95% sensitivity, 487V yielded a 94.9% specificity compared to 86.1% for 487A. EBNA1 deleted for the conserved glycine-alanine repeats (GAr) reduced false positives by 22.8%. NPC sera reacted more strongly to the C-terminus than healthy controls, but the C-terminal construct (a.a. 390-641) showed lower specificity (84.8%) than the EBNA1 GAr deleted construct (92.4%) at 95% sensitivity.
CONCLUSION
Although EBNA1 IgA was present in healthy sera, most epitopes localized to the immunodominant GAr. We conclude that a refined EBNA1 antigen deleted for the GAr but with residues consistently detected in Southeast Asian NPC tumors is optimal for risk prediction with an extended sojourn time of 7.5 years. Furthermore, distinct EBNA1 serologic profiles enhanced the utility of the EBNA1 IgA assay for risk stratification. This illustrates the importance of serologically relevant EBNA1 sequences for NPC risk prediction and early detection.
期刊介绍:
Clinical Cancer Research is a journal focusing on groundbreaking research in cancer, specifically in the areas where the laboratory and the clinic intersect. Our primary interest lies in clinical trials that investigate novel treatments, accompanied by research on pharmacology, molecular alterations, and biomarkers that can predict response or resistance to these treatments. Furthermore, we prioritize laboratory and animal studies that explore new drugs and targeted agents with the potential to advance to clinical trials. We also encourage research on targetable mechanisms of cancer development, progression, and metastasis.