IMMUNOBIOCHEMICAL CHARACTERIZATION WITH MONOCLONAL-ANTIBODIES OF EPSTEIN-BARR VIRUS-ASSOCIATED EARLY ANTIGENS IN CHEMICALLY-INDUCED CELLS
IMMUNOBIOCHEMICAL CHARACTERIZATION WITH MONOCLONAL-ANTIBODIES OF EPSTEIN-BARR VIRUS-ASSOCIATED EARLY ANTIGENS IN CHEMICALLY-INDUCED CELLS
复制标题
DOI：
10.1128/jvi.50.2.372-379.1984
复制
发表时间：
1984-01-01
影响因子：
5.4
通讯作者：
EPSTEIN, AL
中科院分区：
文献类型：
--
作者：
EPSTEIN, AL
五个[小鼠]单克隆抗体，这是反应的早期抗原的EB病毒，通过使用体细胞杂交技术。通过间接免疫荧光证实了单克隆抗体对早期抗原的特异性，其显示抗原定位于早期抗原诱导的[人淋巴母细胞] Raji细胞的细胞核。另外的间接免疫荧光研究表明，像患者抗血清扩散染色早期抗原，单克隆抗体甲醇固定后阳性染色反应。其中一种抗体1150-4通过抗补体免疫荧光技术呈阳性，但与EB病毒相关核抗原阳性患者血清不同，因为它只染色诱导细胞。发现不同的固定方法显著改变了单克隆抗体产生的核染色反应的外观。免疫沉淀和免疫印迹实验显示，单克隆抗体1108-1和1129-1识别55，000和50，000道尔顿（p55;50）的2种多肽; 1173-6和1180-2仅识别p50，1150-4识别65，000道尔顿的核蛋白。这些病毒抗原的免疫生物化学表征表明，p55是一个磷蛋白和p55;50具有较强的DNA结合活性，优先单链DNA。阐明这些核蛋白在EB病毒感染中的作用以及与EB病毒定向淋巴细胞转化相关的事件可能为这种重要的人类病毒的致病性提供重要信息。
Five [mouse] monoclonal antibodies which were reactive to early antigens of Epstein-Barr virus were produced by using somatic cell hybridization techniques. The specificity of the monoclonal antibodies to early antigens was demonstrated by indirect immunofluorescence, which showed that the antigens were localized to the nucleus of early antigen-induced [human lymphoblastoid] Raji cells. Additional indirect immunofluorescence studies showed that like patient antisera to diffuse-staining early antigen, the monoclonal antibodies gave positive staining reactions after methanol fixation. One of the antibodies, 1150-4, was positive by the anticomplement immunofluorescence technique but differed with Epstein-Barr virus-associated nuclear antigen-positive patient sera in that it only stained induced cells. Different fixation methods were found to alter dramatically the appearance of the nuclear staining reactions produced by the monoclonal antibodies. Immunoprecipitation and immunoblot experiments revealed that monoclonal antibodies 1108-1 and 1129-1 recognized 2 polypeptides of 55,000 and 50,000 daltons (p55;50); 1173-6 and 1180-2 recognized just p50 and 1150-4 identified a 65,000 dalton nuclear protein. Immunobiochemical characterization of these viral antigens showed that p55 is a phosphoprotein and p55;50 has strong DNA-binding activity preferentially to single-stranded DNA. Elucidation of the role of these nuclear proteins in Epstein-Barr virus infection and the events associated with Epstein-Barr virus-directed lymphocyte transformation may provide significant information on the pathogenicity of this important human virus.