CHARACTERIZATION OF MONOCLONAL-ANTIBODIES REACTIVE TO SEVERAL BIOCHEMICALLY DISTINCT HUMAN CYTOMEGALOVIRUS GLYCOPROTEIN COMPLEXES
CHARACTERIZATION OF MONOCLONAL-ANTIBODIES REACTIVE TO SEVERAL BIOCHEMICALLY DISTINCT HUMAN CYTOMEGALOVIRUS GLYCOPROTEIN COMPLEXES
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DOI：
10.1128/jvi.60.2.345-352.1986
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发表时间：
1986-11-01
影响因子：
5.4
通讯作者：
GEHRZ, R
中科院分区：
文献类型：
--
作者：
KARI, B;LUSSENHOP, N;GEHRZ, R
三种单克隆抗体的特点是通过检查他们的反应性，人巨细胞病毒（HCMV）糖蛋白在还原和非还原条件下，其反应性的糖蛋白和二硫键连接的糖蛋白复合物分离的离子交换高效液相色谱。一种单克隆抗体9E10与分子量为93，000和450，000的糖蛋白复合物反应，并分别在0.3和0.9 M NaCl下从离子交换柱洗脱。在这些复合物中的所有糖蛋白可以在还原条件下由9E10免疫沉淀，表明它们彼此相关。通过9E10免疫沉淀的最丰富的糖蛋白具有50，000至52，000的分子量。与该抗体相反，另外两种单克隆抗体9B7和41C2与分子量为130，000和> 200，000的糖蛋白复合物反应，并在0.6 M NaCl下从离子交换柱洗脱。在还原条件下，这些复合物中的所有糖蛋白都可以被9B7或41C2免疫沉淀，这表明它们也彼此相关。由41C2或9B7免疫沉淀的最丰富的糖蛋白具有93，000的分子量。此外，还确定了不通过二硫键与其他糖蛋白结合的93，000分子量的糖蛋白不能被三种抗体中的任何一种沉淀，这表明它与其他糖蛋白不同。还检查了单克隆抗体的特异性和中和活性。单克隆抗体41C2和9B7是特异性的HCMV通过免疫荧光染色的皮肤成纤维细胞感染几种不同的病毒。然而，41C2不能中和Towne株HCMV，而9B7可以。9B7的中和活性确实需要称赞。这些结果表明，41C2和9B7与相同糖蛋白上的不同抗原位点反应。与41C2和9B7不同，发现单克隆抗体9E10与腺病毒和单纯疱疹病毒交叉反应，如通过感染的皮肤成纤维细胞的免疫荧光染色所确定的。此外，9E10在不存在补体的情况下中和HCMV的Towne和Toledo株。
Three monoclonal antibodies were characterized by examining their reactivity to human cytomegalovirus (HCMV) glycoproteins under reducing and nonreducing conditions and their reactivity to glycoproteins and disulfide-linked glycoprotein complexes isolated by ion-exchange high-performance liquid chromatography. One monoclonal antibody, 9E10, reacted with glycoprotein complexes which had molecular weights of 93,000 and 450,000 and eluted from the ion-exchange column at 0.3 and 0.9 M NaCl, respectively. All glycoproteins associated in these complexes could be immunoprecipitated under reducing conditions by 9E10, suggesting that they were related to one another. The most abundant glycoproteins immunoprecipitated by 9E10 had molecular weights of 50,000 to 52,000. In contrast to this antibody, two other monoclonal antibodies, 9B7 and 41C2, reacted with glycoprotein complexes which had molecular weights of 130,000 and > 200,000 and eluted from the ion-exchange column at 0.6 M NaCl. All glycoproteins associated in these complexes could be immunoprecipitated by 9B7 or 41C2 under reducing conditions, suggesting that they were also related to one another. The most abundant glycoprotein immunoprecipitated by 41C2 or 9B7 had a molecular weight of 93,000. In addition, it was also determined that a 93,000-molecular-weight glycoprotein which was not associated with other glycoproteins by disulfide bonds could not be precipitated by any of the three antibodies, suggesting that it was different from the other glycoproteins. The monoclonal antibodies were also examined for specificity and neutralizing activity. Monoclonal antibodies 41C2 and 9B7 were specific to HCMV as determined by immunofluorescent staining of skin fibroblast cells infected with several different viruses. However, 41C2 did not neutralize Towne strain HCMV, while 9B7 did. The neutralizing activity of 9B7 did require compliment. These results suggested that 41C2 and 9B7 reacted with different antigenic sites on the same glycoproteins. Unlike 41C2 and 9B7, monoclonal antibody 9E10 was found to cross-react with adenovirus and herpes simplex virus as determined by immunofluorescent stainging of infected skin fibroblast cells. Furthermore, 9E10 neutralized the Towne and Toledo strains of HCMV in the absence of complement.