Antigenic and immunogenic characteristics of nuclear and membrane-associated simian virus 40 tumor antigen.
Antigenic and immunogenic characteristics of nuclear and membrane-associated simian virus 40 tumor antigen.
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核和膜相关猿病毒 40 肿瘤抗原的抗原和免疫原性特征。
DOI：
10.1128/jvi.33.2.887-901.1980
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发表时间：
1980
影响因子：
5.4
通讯作者：
Butel,JS
中科院分区：
文献类型：
--
作者：
Soule,HR;Lanford,RE;Butel,JS
在同基因宿主中制备了针对猴40病毒(SV40)转化细胞亚细胞部分(MoαPM、MoαNuc)、戊二醛固定的SV40转化细胞(HaαH-50-G、MoαVLm-G)和电泳法纯化的变性SV40肿瘤抗原(T-ag)(RaαT)的抗血清。免疫血清来自Sv40转化细胞诱导的荷瘤动物(HaαT、MoαT、HAF)和免疫排斥Sv40转化细胞移植的动物(HaαS、MoαS)。针对细胞表面制备的免疫试剂(MoαPM、HaαS、MoαS、HaαH-50-G、MoαVLm-G)通过间接免疫荧光法或蛋白A表面抗原放射免疫法与转化细胞表面发生特异性反应。针对核组分(MoαNuc)或全细胞决定簇(HaαT、MoαT、HAF、RaαT)制备的免疫试剂可与病毒转化或感染细胞的核和表面反应。所有试剂均能从SV40转化细胞的可溶性全细胞提取液中免疫沉淀96,000个分子量的大T-Ag。HaαS在免疫荧光试验中表现出的独有表面反应性被亚细胞组分的增溶作用所消除，从而允许HaαS从核和质膜标本中免疫沉淀T-Ag。特异性是因为所有的T反应试剂在抗化学转化的小鼠细胞的血清学试验中都不能反应，而移植了化学转化的小鼠细胞(Moαdmba-2)的小鼠的血清也不能与SV40转化的小鼠或仓鼠细胞反应。抗SV40转化细胞的表面免疫荧光阳性和蛋白A放射免疫反应阳性的试剂在双抗体表面抗原放射免疫分析中能够阻断RaαT与SV40转化细胞的表面结合。这种封闭能力直接证明了每个表面反应试剂的成分特异性是针对SV40T-Ag的。提出了一个模型，假设不同血清学试剂对T-Ag的差异检测反映了定位于细胞核和质膜上的T-Ag多肽之间的免疫原性和抗原性差异。
Antisera were prepared in syngeneic hosts against subcellular fractions of simian virus 40 (SV40)-transformed cells (MoαPM, MoαNuc), glutaraldehydefixed SV40-transformed cells (HaαH-50-G, MoαVLM-G), and electrophoretically purified denatured SV40 tumor antigen (T-ag) (RaαT). Immune sera were also collected from animals bearing tumors induced by SV40-transformed cells (HaαT, MoαT, HAF) and from SV40-immunized animals that had rejected a transplant of SV40-transformed cells (HaαS, MoαS). Immunological reagents prepared against cell surface (MoαPM, HaαS, MoαS, HaαH-50-G, MoαVLM-G) reacted exclusively with the surface of SV40-transformed cells by indirect immunofluorescence or protein A surface antigen radioimmunoassay. Immunological reagents prepared against the nuclear fraction (MoαNuc) or whole-cell determinants (HaαT, MoαT, HAF, RaαT) reacted with both the nuclei and surface of SV40-transformed or -infected cells. All reagents were capable of immunoprecipitating 96,000-molecular weight large T-ag from solubilized whole cell extracts of SV40-transformed cells. The exclusive surface reactivity of HaαS exhibited in immunofluorescence tests was abolished by solubilization of subcellular fractions, which then allowed immunoprecipitation of T-ag by HaαS from both nuclear and plasma membrane preparations. Specificity was established by the fact that all T-reactive reagents failed to react in serological tests against chemically transformed mouse cells, and sera from mice bearing transplants chemically transformed mouse cells (MoαDMBA-2) failed to react with SV40-transformed mouse or hamster cells. Reagents demonstrating positive surface immunofluorescence and protein A radioimmunoassay reactions against SV40-transformed cells were capable of blocking the surface binding of RaαT to SV40-transformed cells in a double-antibody surface antigen radioimmunoassay. This blocking ability demonstrated directly that a component specificity of each surface-reactive reagent is directed against SV40 T-ag. A model is presented which postulates that the differential detection of T-ag by the various serological reagents is a reflection of immunogenic and antigenic differences between T-ag polypeptides localized in nuclei and plasma membranes.