Use of bacterially expressed GST/EBNA-1 fusion proteins for detection of antibodies in sera from patients with nasopharyngeal carcinoma and healthy donors.

M R Chen, J F Yang, T Y Hsu, M Y Liu, J Y Chen, C S Yang
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Abstract

Epstein-Barr virus nuclear antigen-1 (EBNA-1) is a protein expressed consistently in EBV infected cells and in EBV related malignant tissues. Antibodies against EBNA-1 may therefore possibly be used as a marker for disease screening. Western blot analysis of serum antibodies was performed using GST (glutathione-S-transferase) fusion proteins containing different regions of EBNA-1 as antigens. Serum samples were collected from 38 patients with nasopharyngeal carcinoma (NPC) and 38 healthy individuals in Taiwan. All samples were found IgG positive for EBNA-1 when a truncated protein GST/E1 (70-102, 325-641) was used as the antigen. Thirty-three out of 38 NPC sera (86.8%) were positive for IgA antibody against EBNA-1. The positive rate was higher in comparison with IgA antibody against VCA (65.7%) or antibody against DNase (60.5%). Only 2.6% of sera from normal individuals were positive for an IgA response against EBNA-1. The major antigenic determinants for NPC serum IgA response were between amino acid(aa) 390 to aa 459 when different portions of EBNA-1 were used as antigens. The results suggest that IgA response against EBNA-1 could be used in combination with other EBV serology markers for NPC screening.

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利用细菌表达的GST/EBNA-1融合蛋白检测鼻咽癌患者和健康供者血清中的抗体
eb病毒核抗原-1 (EBNA-1)是一种在EBV感染细胞和EBV相关恶性组织中一致表达的蛋白。因此,针对EBNA-1的抗体可能用作疾病筛查的标记物。采用含有EBNA-1不同区域的GST(谷胱甘肽- s -转移酶)融合蛋白作为抗原进行血清抗体的Western blot分析。本文收集了台湾地区38例鼻咽癌患者和38例正常人的血清样本。用截断的GST/E1蛋白(70- 102,325 -641)作为抗原,所有样本均发现EBNA-1 IgG阳性。38份NPC血清中有33份(86.8%)EBNA-1 IgA抗体阳性。IgA抗VCA抗体阳性率为65.7%,DNase抗体阳性率为60.5%。正常人血清中只有2.6%的人对EBNA-1的IgA反应呈阳性。当EBNA-1的不同部分作为抗原时,鼻咽癌血清IgA反应的主要抗原决定因子在氨基酸(aa) 390 ~ aa 459之间。结果表明,IgA对EBNA-1的反应可与其他EBV血清学标志物联合用于鼻咽癌筛查。
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