Lin P S, Schmidt-Ullrich R, Wallach D F
Proc Natl Acad Sci U S A. 1977 Jun;74(6):2495-9. doi: 10.1073/pnas.74.6.2495.
Nucleus- and mitochondrion-free membranes from hamster lymphocytes transformed by simian virus 40 (SV40), GD248 cells, cause guinea pigs to produce immune sera that reveal the presence in GD248 plasma membranes and mitochondria of two types of glycoprotein that are not detected in membranes of normal lymphocytes [Schmidt-Ullrich, R., Thompson, W. S. & Wallach, D. F. H. (1977) Proc. Natl. Acad. Sci. USA 74, 643-647]. Indirect immune fluorescence of living, SV40-transformed T19 hamster reticulum cells, Balb/c 3T3 mouse fibroblasts, and W18 VA2 human fibroblasts, using the antisera against GD248 membrane, at 4 degrees produced a distinct cell surface fluorescence; however, above 20 degrees , staining at the nuclear perimeter, the SV40 U-antigen reaction, becomes equally prominent. In SV40-transformed cells that had been fixed in cold acetone, as well as in purified GD248 nuclei, thermostable U-antigen staining is dramatic, but there is no reaction for nuclear T-antigen. Rabbit antisera against T19 cells gave immunofluorescence reactions equivalent to those obtained with the antisera against GD248 cells. Normal guinea pig or rabbit sera and cells that had not been transformed by SV40 gave no reaction. Our sera from tumor-bearing hamsters gave only nuclear T-antigen fluorescence. The results indicate the presence of related, SV40-specific antigens in the surface membranes, nuclear envelope, and possibly other intracellular organelles of SV40-transformed cells.
由猿猴病毒40(SV40)转化的仓鼠淋巴细胞(GD248细胞)中不含细胞核和线粒体的膜,能使豚鼠产生免疫血清,该血清显示在GD248细胞膜和线粒体中存在两种在正常淋巴细胞膜中未检测到的糖蛋白[施密特 - 乌尔里希,R.,汤普森,W. S. & 瓦拉赫,D. F. H.(1977年)美国国家科学院院刊74,643 - 647]。使用针对GD248膜的抗血清,在4℃对活的、SV40转化的T19仓鼠网状细胞、Balb/c 3T3小鼠成纤维细胞和W18 VA2人成纤维细胞进行间接免疫荧光检测,产生了明显的细胞表面荧光;然而,在20℃以上,核周染色,即SV40 U抗原反应,变得同样明显。在冷丙酮中固定的SV40转化细胞以及纯化的GD248细胞核中,热稳定的U抗原染色很显著,但对核T抗原则无反应。针对T19细胞的兔抗血清产生的免疫荧光反应与针对GD248细胞的抗血清所获得的反应相当。未被SV40转化的正常豚鼠或兔血清以及细胞无反应。我们从荷瘤仓鼠获得的血清仅产生核T抗原荧光。结果表明在SV40转化细胞的表面膜、核膜以及可能的其他细胞内细胞器中存在相关的、SV40特异性抗原。