Cianciolo G J, Lostrom M E, Tam M, Snyderman R
J Exp Med. 1983 Sep 1;158(3):885-900. doi: 10.1084/jem.158.3.885.
Murine tumors contain low molecular weight factors that inhibit macrophage accumulation at inflammatory foci. Certain oncogenic murine leukemia viruses contain similar inhibitory activity and the active component of the retroviruses was shown to be the envelope protein P15E. A number of murine malignant and nonmalignant cell lines, as well as primary tumors, have now been examined to determine whether production of retroviral P15E or a related protein is characteristic of neoplastic cells. Tumor lines examined included the Hep 129 hepatocarcinoma, BP8 fibrosarcoma, RL1 lymphoma, and three variants of the B16 melanoma. Tumor lines were virus negative by electron microscopy. Nonmalignant cells examined included ST0, 3T3/BALB, and 3T3/L1 fibroblasts and unstimulated, as well as mitogen-stimulated murine splenocytes. Cells were pulse-labeled with [35S]methionine, proteins immunoprecipitated with two monoclonal antibodies to P15E and analyzed by SDS-PAGE and gel fluorography. All tumor lines synthesized a approximately 19,000-dalton protein that co-migrated with retroviral P15E on SDS-PAGE. None of the nonmalignant cells synthesized this protein. Two-dimensional gel electrophoresis of the proteins precipitated from two B16 melanoma lines by monoclonal anti-P15E showed them to be physicochemically similar to P15E from Rauscher leukemia virus. A competition ELISA assay for P15E was developed and confirmed the results obtained by metabolic labeling and demonstrated P15E-related antigens in the tumor cell lines and also in the ascites fluid of mice injected with Hep 129 cells. More importantly, P15E antigens were expressed in both a spontaneous mammary adenocarcinoma and in a primary methylcholanthrene-induced fibrosarcoma. Nonmalignant tissues from animals bearing these tumors contained no detectable P15E antigen. Extracts from the primary fibrosarcomas, when injected into the thighs of mice, inhibited the intraperitoneal accumulation of inflammatory macrophages. The inhibitory activity was specifically removed by absorption with monoclonal antibody to P15E. These results suggest that synthesis of the immunosuppressive retroviral protein P15E, or a very similar protein, routinely occurs during the growth of murine neoplastic cells. This P15E-related protein is present in spontaneous murine primary tumors as well as in all murine tumor cell lines tested. The expression of such proteins by transformed cells in vivo could confer a selective advantage for their sustained growth since they would be more likely to escape immune surveillance.
小鼠肿瘤含有低分子量因子,可抑制巨噬细胞在炎症灶的聚集。某些致癌性小鼠白血病病毒具有类似的抑制活性,并且逆转录病毒的活性成分被证明是包膜蛋白P15E。现在已经检测了许多小鼠恶性和非恶性细胞系以及原发性肿瘤,以确定逆转录病毒P15E或相关蛋白的产生是否是肿瘤细胞的特征。检测的肿瘤细胞系包括Hep 129肝癌、BP8纤维肉瘤、RL1淋巴瘤以及B16黑色素瘤的三个变体。通过电子显微镜检查,肿瘤细胞系均未检测到病毒。检测的非恶性细胞包括ST0、3T3/BALB和3T3/L1成纤维细胞,以及未受刺激和经丝裂原刺激的小鼠脾细胞。用[35S]甲硫氨酸对细胞进行脉冲标记,用两种抗P15E单克隆抗体免疫沉淀蛋白质,然后通过SDS-PAGE和凝胶荧光成像进行分析。所有肿瘤细胞系均合成了一种分子量约为19,000道尔顿的蛋白质,该蛋白质在SDS-PAGE上与逆转录病毒P15E迁移位置相同。非恶性细胞均未合成这种蛋白质。用单克隆抗P15E抗体沉淀自两个B16黑色素瘤细胞系的蛋白质进行二维凝胶电泳,结果显示它们在物理化学性质上与劳氏肉瘤病毒的P15E相似。开发了一种针对P15E的竞争ELISA检测方法,证实了代谢标记获得的结果,并证明肿瘤细胞系以及注射了Hep 129细胞的小鼠腹水中存在P15E相关抗原。更重要的是,P15E抗原在自发性乳腺腺癌和原发性甲基胆蒽诱导的纤维肉瘤中均有表达。携带这些肿瘤的动物的非恶性组织未检测到可检测到的P15E抗原。原发性纤维肉瘤的提取物注射到小鼠大腿后,可抑制炎症巨噬细胞在腹腔内的聚集。用抗P15E单克隆抗体吸附可特异性去除抑制活性。这些结果表明,免疫抑制性逆转录病毒蛋白P15E或非常相似的蛋白在小鼠肿瘤细胞生长过程中经常合成。这种与P15E相关的蛋白存在于自发性小鼠原发性肿瘤以及所有测试的小鼠肿瘤细胞系中。体内转化细胞表达此类蛋白可能为其持续生长赋予选择性优势,因为它们更有可能逃避免疫监视。