Epitope analysis of human cytomegalovirus glycoprotein complexes using murine monoclonal antibodies.
Epitope analysis of human cytomegalovirus glycoprotein complexes using murine monoclonal antibodies.
复制标题
使用鼠单克隆抗体对人巨细胞病毒糖蛋白复合物进行表位分析。
DOI：
10.1016/0042-6822(88)90549-1
复制
发表时间：
1988
期刊：
影响因子：
3.7
通讯作者：
Kari,B
中科院分区：
文献类型：
--
作者：
Lussenhop,NO;Goertz,R;Wabuke-Bunoti,M;Gehrz,R;Kari,B
生成了一组 10 种与人巨细胞病毒 (HCMV) 糖蛋白反应的单克隆抗体。这些抗体免疫沉淀二硫键连接的复合物，其中含有分子量为 130,000、93,000 和 52,000 的糖蛋白。这些复合物被命名为gC-I。 gC-I 的表位分析通过同时进行的两种抗体结合测定进行。揭示了一个表位网络，该网络聚集在三个主要域中，分别称为 I、II 和 III。各个结构域 I 和 II 内的抗体表现出对彼此结合的强烈相互抑制。然而，结构域 I 和结构域 II 之间存在多种抗体相互作用。例如，结构域 I 中大多数抗体的结合在某种程度上被结构域 II 中的抗体增强。然而，仅来自结构域 II 的一种抗体的结合被来自结构域 I 的所有抗体增强。两种抗体之间的结合增强取决于 gC-I 的天然结构，并且对由于 gC-I 的非离子去污剂提取和/或二硫键破坏而导致的构象变化敏感。在病毒中和试验中，域 I 和 II 中的抗体之间也观察到了协同效应。在非中和抗体存在的情况下，中和抗体具有大得多的中和活性，这也增强了同时两种抗体结合测定中中和抗体的结合。此外，两种单独具有非中和性的抗体在组合使用时具有中和性。在同时进行的两种抗体结合测定中，此类抗体还增强了彼此的结合。最后，结构域 III 由非中和抗体组成，抑制结构域 I 和 II 中所有抗体的结合。该抗体还在病毒中和测定中抑制中和抗体的中和活性。
A panel of 10 monoclonal antibodies reactive with human cytomegalovirus (HCMV) glycoproteins was generated. These antibodies immunoprecipitated disulfide-linked complexes which contained glycoproteins with molecular weights of 130,000, 93,000, and 52,000. These complexes were designated gC-I. Epitope analysis of gC-I was done with a simultaneous two antibody binding assay. A network of epitopes was revealed which clustered in three major domains designated I, II, and III. Antibodies within individual domains I and II showed strong mutual inhibition of each other's binding. However, there were multiple antibody interactions between domains I and II. For example, the binding of most antibodies in domain I was augmented to some extent by antibodies from domain II. However, the binding of only one antibody from domain II was augmented by all antibodies from domain I. The augmentation in binding between two antibodies was dependent on the native structure of gC-I and was sensitive to conformational changes due to nonionic detergent extraction of gC-I and/or disruption of disulfide bonds. A synergistic effect was also observed between antibodies in domains I and II in a virus neutralization assay. A neutralizing antibody had a much greater neutralizing activity in the presence of a nonneutralizing antibody, which also enhanced the binding of the neutralizing antibody in the simultaneous two antibody binding assay. Also, two antibodies which were nonneutralizing individually were neutralizing when used in combination. Such antibodies also augmented each other's binding in the simultaneous two antibody binding assay. Finally, domain III consisted of a nonneutralizing antibody that inhibited the binding of all antibodies in domains I and II. This antibody also inhibited the neutralizing activity of a neutralizing antibody in a virus neutralizing assay.