Monoclonal antibodies to human cytomegalovirus: three surface membrane proteins with unique immunological and electrophoretic properties specify cross-reactive determinants
Monoclonal antibodies to human cytomegalovirus: three surface membrane proteins with unique immunological and electrophoretic properties specify cross-reactive determinants
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人巨细胞病毒单克隆抗体：具有独特免疫学和电泳特性的三种表面膜蛋白指定交叉反应决定簇
作者：
L. Pereira;M. Hoffman;D. Gallo;N. Cremer
以人巨细胞病毒(CMV)AD169株免疫小鼠脾细胞与小鼠骨髓瘤细胞融合，经免疫荧光筛选获得77个与CMV感染细胞有反应的杂交瘤细胞克隆。根据从CMV感染细胞提取液中免疫沉淀的多肽的电泳性，将克隆分为七组。针对CMV表膜抗原的三组单抗的研究结果如下：根据免疫球蛋白亚类、中和活性和与暴露在完整CMV感染细胞表膜上的蛋白的抗原域的反应性来区分每组克隆。每组中的单抗沉淀出一种慢速迁移蛋白和多种快速迁移形式，它们具有相同的抗原决定簇。第一组单抗沉淀了4个糖基化多肽，表观分子量分别为130,000,110,000,100,000和60,000。该组单抗CH51能中和感染性病毒，但不能与感染细胞表面的抗原区发生反应。第二组单抗沉淀了四个表观分子量分别约为66,000，55,000，50,000和46,000的多肽。该组中的单抗CH65和CH134具有中和活性，并与暴露在CMV感染细胞表面的蛋白的抗原域发生反应。第三组单抗沉淀了4个表观分子量分别为49,000，48,000，34,000和25,000的多肽。用一组抗CMV膜蛋白的单抗对15个自然发生的CMV株系进行了血清学分析，结果表明，所有株系与所测抗体反应的抗原决定簇都是保守的。抗CMV感染细胞表面膜蛋白的单抗可能成为鉴定病毒分离物的有价值的试剂。
Seventy-seven clones of hybridomas selected for reactivity by immunofluorescence with human cytomegalovirus (CMV)-infected cells were produced by fusing mouse myeloma cells with the spleen cells of mice immunized with CMV strain AD169. The clones were classified into seven groups on the basis of the electrophoretic properties of the polypeptides immune precipitated from extracts of CMV-infected cells. Studies on the three groups of monoclonal antibodies directed against CMV surface membrane antigens showed the following. Clones in each group were differentiated by immunoglobulin subclass, neutralizing activity, and reactivity with the antigenic domains of proteins exposed on the surface membranes of intact CMV-infected cells. Monoclonal antibodies in each group precipitated one slowly migrating protein and multiple faster migrating forms which shared antigenic determinants. The first group of monoclonal antibodies precipitated four glycosylated polypeptides with apparent molecular weights of 130,000, 110,000, 100,000, and 60,000. Monoclonal antibody CH51 of this group neutralized infectious virus but failed to react with antigenic domains on the surfaces of infected cells. The second group of monoclonal antibodies precipitated four polypeptides with apparent molecular weights of approximately 66,000, 55,000, 50,000, and 46,000. Monoclonal antibodies CH65 and CH134 in this group had neutralizing activity and reacted with antigenic domains of proteins exposed on the surface of CMV-infected cells. The third group of monoclonal antibodies precipitated four polypeptides with apparent molecular weights of 49,000, 48,000, 34,000, and 25,000. Serological analysis of 15 naturally occurring CMV strains with a panel of monoclonal antibodies to surface membrane proteins showed that the antigenic determinants reactive with the antibodies tested were conserved in all of the strains. Monoclonal antibodies to surface membrane proteins on CMV-infected cells may prove to be valuable reagents for identification of virus isolates.