Lenstra J A, Kusters J G, Koch G, van der Zeijst B A
Institute of Infectious Diseases and Immunology, Veterinary Faculty, State University, Utrecht, The Netherlands.
Mol Immunol. 1989 Jan;26(1):7-15. doi: 10.1016/0161-5890(89)90014-x.
To study the antigenic structure of the peplomer protein of the avian coronavirus infectious bronchitis virus, fragments from the peplomer gene were generated by restriction-enzyme cleavage or by limited DNase digestion and inserted in the Escherichia coli expression plasmid pEX (Stanley and Luzio, 1984). The antigenicity of the expression products was tested using a number of polyclonal antisera and monoclonal antibodies. The polyclonal antisera recognized different sets of epitopes in the 1162-residue sequence. The N-terminal region of one of the two subunits, S2, was recognized by all polyclonal sera and by two monoclonal antibodies. This clearly immunodominant region contains at least two adjacent or overlapping epitopes, one of which has been localized within 18 residues. The epitopes found as antigenic pEX expression products do not coincide with the regions in the S1 subunit that have been found to contain hypervariable sequences. We suggest that these regions constitute conformation dependent neutralization epitopes that cannot be detected in the pEX system. The relevance of our findings for vaccine development is discussed.
为研究禽冠状病毒传染性支气管炎病毒纤突蛋白的抗原结构，通过限制性酶切或有限的脱氧核糖核酸酶消化产生纤突基因片段，并将其插入大肠杆菌表达质粒pEX（斯坦利和卢齐奥，1984年）。使用多种多克隆抗血清和单克隆抗体检测表达产物的抗原性。多克隆抗血清在1162个残基序列中识别不同的表位组。两个亚基之一S2的N端区域被所有多克隆血清和两种单克隆抗体识别。这个明显的免疫显性区域至少包含两个相邻或重叠的表位，其中之一已定位在18个残基范围内。作为抗原性pEX表达产物发现的表位与已发现含有高变序列的S1亚基区域不一致。我们认为这些区域构成构象依赖性中和表位，在pEX系统中无法检测到。讨论了我们的发现对疫苗开发的相关性。
