Monoclonal antibodies against simian virus 40 T antigens: evidence for distinct sublcasses of large T antigen and for similarities among nonviral T antigens
Monoclonal antibodies against simian virus 40 T antigens: evidence for distinct sublcasses of large T antigen and for similarities among nonviral T antigens
复制标题
抗猿猴病毒 40 T 抗原的单克隆抗体：大 T 抗原不同亚型以及非病毒 T 抗原之间相似性的证据
作者：
E. Gurney;R. Harrison;J. Fenno
我们已经分离出三个克隆的杂交细胞，合成抗体的决定簇对猿猴病毒40（SV 40）的T抗原。将小鼠骨髓瘤NS 1细胞与来自用SV 40转化的小鼠细胞免疫的小鼠的脾细胞融合。在HAT培养基中选择杂交细胞并在软琼脂中克隆。我们使用酶联免疫吸附试验检测和定量小鼠抗SV 40 T抗原的抗体。通过免疫沉淀法验证了在酶联免疫吸附试验中评分为阳性的24个克隆中3个克隆的单克隆抗体对SV 40 T抗原具有特异性。两个克隆（7和412）产生的抗体，识别变性敏感的抗原决定簇独特的大T抗原。来自克隆7的抗体似乎对大T抗原具有低亲和力。来自克隆412的抗体对大T抗原具有更高的亲和力，但不识别被肿瘤血清识别的大T抗原的亚类。第三个克隆（克隆122）的抗体识别SV 40转化细胞中53，000-道尔顿小鼠非病毒T抗原的变性稳定抗原决定簇。克隆122的抗体也能识别来自5种哺乳动物的SV 40转化或裂解感染细胞和4种未感染小鼠品系的相似（51，000 - 56，000-道尔顿）非病毒T抗原。根据这些观察，我们得出结论：（i）94，000-道尔顿的SV 40大T抗原可能作为免疫学上可区分的亚类存在，（ii）5种哺乳动物的非病毒T抗原至少共享一个抗原决定簇。
We have isolated three clones of hybrid cells which synthesize antibodies specific for determinants on simian virus 40 (SV40) T antigens. Mouse myeloma NS1 cells were fused with spleen cells from mice that had been immunized with SV40-transformed mouse cells. Hybrid cells were selected in HAT medium and cloned in soft agar. We used an enzyme-linked immunosorbent assay for detection and quantification of mouse antibodies against SV40 T antigens. Monoclonal antibodies from 3 of the 24 clones that scored as positive in the enzyme-linked immunosorbent assay were verified by immunoprecipitation to be specific for SV40 T antigens. Two clones (7 and 412) produced antibodies that recognized denaturation-sensitive antigenic determinants unique to large T antigen. Antibodies from clone 7 appeared to have a low affinity for large T antigen. Antibodies from clone 412 had a higher affinity for large T antigen but did not recognize a subclass of large T antigen that was recognized by tumor serum. Antibodies of the third clone, clone 122, recognized a denaturation-stable antigenic determinant of the 53,000-dalton mouse nonviral T antigen in SV40-transformed cells. Antibodies from clone 122 also recognized similar (51,000- to 56,000-dalton) nonviral T antigens in SV40-transormed or lytically infected cells from five mammalian species and in four uninfected mouse lines. From these observations, we have concluded that (i) the 94,000-dalton SV40 large T antigen may exist as immunologically distinguishable subclasses, and (ii) the nonviral T antigens of five mammalian species share at least one antigenic determinant.