Fowler W J, Garcia-Valcarcel M, Hill-Perkins M S, Murphy G, Harper D R, Jeffries D J, Burns N R, Adams S E, Kingsman A J, Layton G T
British Biotech Pharmaceuticals Ltd., Oxford, United Kingdom.
Virology. 1995 Dec 20;214(2):531-40. doi: 10.1006/viro.1995.0064.
The envelope proteins of varicella-zoster virus (VZV) are highly immunogenic and one of the most abundant is glycoprotein E (gE). However, its immunodominant regions and epitopes have not been identified. In this study, using human sera from individuals with recent varicella or zoster infections, we have localized antigenic sequences of gE using recombinant hybrid Ty-virus-like particles (VLPs) carrying overlapping fragments of the gE protein. gE(1-134)-VLPs (particles carrying amino acids 1-134 of gE) and, to a lesser extent, gE(101-161)-VLPs were found to be the most antigenic when tested by Western blotting and ELISA. Other fragments of gE (spanning residues 161-623) showed weak or no antigenicity. Pepscan analysis of human sera on overlapping synthetic peptides representing residues 1-135 of gE revealed that the most antigenic region was between residues 50 and 135. Three immunodominant sequences (residues 86-105, 116-135, and, to a lesser extent, 56-75) were detected using sera from both varicella and zoster patients. All sera from varicella, but not zoster, patients reacted strongly with an epitope in peptide 66-85. Other epitopes were recognized weakly by some varicella or zoster sera. More sera need to be tested to assess the potential disease specificity of these epitopes. The neutralizing monoclonal antibody (MAb) IF-B9 reacted with residues 71-90; however, another neutralizing MAb, SG1A, which bound to both gE(1-134)-VLPs and gE(101-161)-VLPs did not bind to any peptide. The identification of immunodominant sequences of gE will help toward the development of a subunit VZV vaccine.
水痘带状疱疹病毒（VZV）的包膜蛋白具有高度免疫原性，其中最丰富的一种是糖蛋白E（gE）。然而，其免疫显性区域和表位尚未确定。在本研究中，我们使用近期患水痘或带状疱疹个体的人血清，利用携带gE蛋白重叠片段的重组杂交Ty病毒样颗粒（VLP）定位了gE的抗原序列。通过蛋白质印迹法和酶联免疫吸附测定法检测发现，gE(1 - 134)-VLP（携带gE第1 - 134位氨基酸的颗粒）以及在较小程度上gE(101 - 161)-VLP具有最强的抗原性。gE的其他片段（跨越第161 - 623位残基）显示出较弱的抗原性或无抗原性。对代表gE第1 - 135位残基的重叠合成肽进行的人血清肽扫描分析表明，最具抗原性的区域在第50至135位残基之间。使用水痘和带状疱疹患者的血清检测到三个免疫显性序列（第86 - 105位残基、第116 - 135位残基以及在较小程度上第56 - 75位残基）。所有水痘患者的血清，但带状疱疹患者的血清未出现这种情况，与肽段66 - 85中的一个表位发生强烈反应。其他表位被一些水痘或带状疱疹血清微弱识别。需要检测更多血清以评估这些表位的潜在疾病特异性。中和单克隆抗体（MAb）IF - B９与第71 - 90位残基发生反应；然而，另一种中和单克隆抗体SG1A，它既能结合gE(1 - 134)-VLP，也能结合gE(101 - 161)-VLP，但不与任何肽段结合。gE免疫显性序列的鉴定将有助于亚单位VZV疫苗的开发。
