Keydar I, Ohno T, Nayak R, Sweet R, Simoni F, Weiss F, Karby S, Mesa-Tejada R, Spiegelman S
Proc Natl Acad Sci U S A. 1984 Jul;81(13):4188-92. doi: 10.1073/pnas.81.13.4188.
Clonal derivatives 8 and 11 of the T47D human breast carcinoma cell line release particles that have the biochemical characteristics of a retrovirus. Particles recovered from cultures of [3H]uridine-labeled clone 11 had a density of 1.18 g/ml and contained 60-70S and 35S RNAs associated with reverse transcriptase activity. The production of these particles was steroid-dependent. Clone 8 particles had a higher density, 1.195 g/ml, and their production was independent of steroid hormone. By RIA, antigens crossreactive with the 52,000-dalton envelope glycoprotein gp52, the major external protein of mouse mammary tumor virus, were found associated with these particles and in the media. Most of the gp52-related antigen was in soluble form, but it was enriched in the particle preparation. A lesser amount of antigen was distributed within the cultured cells. Absorption of rabbit antibody to gp52 with clone 11 particle preparations eliminated the ability of this antibody to detect immunocytochemically a crossreactive antigen previously localized in tissue sections of human breast carcinoma. These results indicate that the particle isolates from T47D contain the same gp52-related antigen found in human breast carcinomas and constitute an excellent source for the purification and characterization of this antigen.
T47D人乳腺癌细胞系的克隆衍生物8和11释放出具有逆转录病毒生化特性的颗粒。从[³H]尿苷标记的克隆11培养物中回收的颗粒密度为1.18 g/ml,含有与逆转录酶活性相关的60 - 70S和35S RNA。这些颗粒的产生是类固醇依赖性的。克隆8的颗粒密度更高,为1.195 g/ml,其产生与类固醇激素无关。通过放射免疫分析(RIA)发现,与小鼠乳腺肿瘤病毒主要外部蛋白52,000道尔顿包膜糖蛋白gp52交叉反应的抗原与这些颗粒及培养基相关。大多数与gp52相关的抗原呈可溶形式,但在颗粒制剂中富集。较少数量的抗原分布在培养细胞内。用克隆11颗粒制剂吸收兔抗gp52抗体后,该抗体无法通过免疫细胞化学方法检测到先前在人乳腺癌组织切片中定位的交叉反应抗原。这些结果表明,从T47D分离出的颗粒含有在人乳腺癌中发现的相同的与gp52相关的抗原,是纯化和鉴定该抗原的极佳来源。