MONOCLONAL-ANTIBODIES TO NEWCASTLE-DISEASE VIRUS - DELINEATION OF 4 EPITOPES ON THE HN GLYCOPROTEIN
MONOCLONAL-ANTIBODIES TO NEWCASTLE-DISEASE VIRUS - DELINEATION OF 4 EPITOPES ON THE HN GLYCOPROTEIN
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DOI：
10.1128/jvi.48.2.440-450.1983
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发表时间：
1983-01-01
影响因子：
5.4
通讯作者：
BRATT, MA
中科院分区：
文献类型：
--
作者：
IORIO, RM;BRATT, MA
用新城疫病毒澳大利亚-维多利亚株紫外线灭活后免疫的BALB/c小鼠的脾淋巴细胞与SP2细胞融合，制备了18株产生抗新城疫病毒单抗的独立杂交瘤细胞。根据放射免疫沉淀、感染鸡胚成纤维细胞表面和胞浆荧光及同型，将其分为3组。抗HN[血凝素神经氨酸酶]组由9个抗体组成，这些抗体在感染细胞上发出表面荧光并免疫沉淀HN糖蛋白。这些抗体在硝基纤维素转移的十二烷基硫酸钠凝胶中与HN结合，但前提是它既没有还原也没有煮熟。所有这些HN抗体都在不同程度上中和了传染性。这些结果表明，它们识别天然HN分子上暴露的构象性质的决定因素。它们已被用于竞争抗体结合放射免疫分析和加法中和分析，在这些研究的基础上，它们识别的表位被细分为4个区域，其中2个区域重叠在HN糖蛋白上。其中一些抗体的中和活性相对较弱，不能用它们表位的亲和力较低来解释，因为估计的结合常数和中和活性之间没有负相关。在还原和未还原的十二烷基硫酸钠-聚丙烯酰胺凝胶的硝酸纤维素转移中，第2组的4种抗体均以胞浆荧光为主，免疫沉淀核衣壳蛋白并与核衣壳蛋白结合。与其他均为免疫球蛋白抗体不同，第3组中的5种抗体均为IgM类抗体。这个组的成员表现出不同的荧光模式，但没有一个能够免疫沉淀或结合从十二烷基硫酸钠-聚丙烯酰胺凝胶转移到硝酸纤维素纸上的特定病毒抗原。
Eighteen independent hybridomas producing monoclonal antibodies to Newcastle disease virus have been prepared by fusion of SP2 cells with spleen lymphocytes from a BALB/c mouse immunized with intact UV-inactivated Newcastle disease virus strain Australia-Victoria. They have been divided into 3 groups on the basis of radioimmunoprecipitation, infected chicken embryo fibroblast cell surface and cytoplasmic fluorescence and isotype. The anti-HN [hemagglutinin-neuraminidase] group is made up of 9 antibodies which give surface fluorescence on infected cells and immunoprecipitate the HN glycoprotein. These antibodies bind to HN in nitrocellulose transfers of sodium dodecyl sulfate gels, but only if it has been neither reduced nor boiled. To varying degrees, all of these HN antibodies neutralize infectivity. These results suggest that they recognize exposed determinants of a conformational nature on the native HN molecule. They have been used in competition antibody-binding radioimmunoassays and additive neutralization assays, and on the basis of these studies the epitopes they recognize have been subdivided into 4 domains, 2 of which are overlapping on the HN glycoprotein. The relatively weaker neutralizing activity observed with some of these antibodies cannot be explained by lower avidities for their epitopes because there is not an inverse correlation between estimated binding constant and neutralizing activity. The 4 antibodies in the 2nd group all give a predominantly cytoplasmic fluorescence pattern, immunoprecipitate the nucleocapsid protein and bind to nucleocapsid protein in nitrocellulose transfers of reduced and nonreduced sodium dodecyl sulfate-polyacrylamide gels. All 5 of the antibodies in the 3rd group are of the IgM class, unlike the others which are all IgG antibodies. Members of this group show variable fluorescence patterns, but none is able to immunoprecipitate or bind to a specific viral antigen transferred to nitrocellulose paper from sodium dodecyl sulfate-polyacrylamide gels.