Antigenic structure of the E2 glycoprotein from transmissible gastroenteritis coronavirus.
Antigenic structure of the E2 glycoprotein from transmissible gastroenteritis coronavirus.
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DOI：
10.1016/0168-1702(88)90059-7
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发表时间：
1988-04
期刊：
影响因子：
5
通讯作者：
Enjuanes L
中科院分区：
文献类型：
--
作者：
Correa I;Jiménez G;Suñé C;Bullido MJ;Enjuanes L
传染性胃肠炎病毒（TGE）E2糖蛋白的抗原结构分为抗原位点、抗原亚位点和表位三个层次。用竞争性放射免疫分析法（RIA）确定了4个抗原位点（A、B、C和D）。约20%（197）的TGE病毒特异性杂交瘤产生了对位点A特异性的中和单克隆抗体，这是抗原显性决定簇之一。位点A被区分为三个抗原亚位点：a、B和c，通过表征11个MAB抗性（mar）突变体，分别由8、3和3个MAbs定义。这些亚位点在表位中进一步细分。在E2糖蛋白中共定义了11个表位，其中8个是病毒中和的关键。仅当使用天然病毒免疫小鼠时才获得中和MAb，尽管为了产生杂交瘤，小鼠免疫是用天然、变性或天然和变性形式的混合物的抗原进行的。所有中和单克隆抗体均与构象表位反应。E2-糖蛋白的抗原结构已经用鼠单克隆抗体确定，但抗原位点在猪（病毒的天然宿主）中是相关的，因为猪血清对这些位点有反应。TGE病毒C位点特异性单克隆抗体与非免疫猪血清反应。该反应性不针对猪免疫球蛋白。这些结果表明，TGE病毒含有的表位也存在于猪血清的一些非免疫球蛋白组分中。
The antigenic structure of transmissible gastroenteritis (TGE) virus E2 glycoprotein has been defined at three levels: antigenic sites, antigenic subsites and epitopes. Four antigenic sites (A, B, C and D) were defined by competitive radioimmunoassay (RIA) using monoclonal antibodies (MAbs) selected from 9 fusions. About 20% (197) of the hybridomas specific for TGE virus produced neutralizing MAbs specific for site A, which was one of the antigenically dominant determinants. Site A was differentiated in three antigenic subsites: a, b and c, by characterization of 11 MAb resistant (mar) mutants, that were defined by 8, 3, and 3 MAbs, respectively. These subsites were further subdivided in epitopes. A total of 11 epitopes were defined in E2 glycoprotein, eight of which were critical for virus neutralization. Neutralizing MAbs were obtained only when native virus was used to immunize mice, although to produce hybridomas mice immunizations were made with antigen in the native, denatured, or mixtures of native and denatured form. All neutralizing MAbs reacted to conformational epitopes. The antigenic structure of the E2-glycoprotein has been defined with murine MAbs, but the antigenic sites were relevant in the swine, the natural host of the virus, because porcine sera reacted against these sites. MAbs specific for TGE virus site C reacted to non-immune porcine sera. This reactivity was not directed against porcine immunoglobulins. These results indicated that TGE virus contains epitope(s) also present in some non-immunoglobulin component of porcine serum.
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