Suda T, Shimizu J, Mizushima Y, Fujiwara H, Hamaoka T
Department of Oncogenesis, Institute for Cancer Research, Osaka.
Jpn J Cancer Res. 1988 Mar;79(3):365-74. doi: 10.1111/j.1349-7006.1988.tb01600.x.
The tumor antigen capable of inducing tumor resistance (tumor rejection antigen; TRA) was separated and some of its physicochemical properties were characterized. Cytosol and plasma membrane fractions were separated from Rous sarcoma virus (RSV)-induced CSA1M tumor cells. Immunization with membrane but not cytosol fraction of these tumor cells together with complete Freund's adjuvant resulted in complete protection against subsequent challenge with viable CSA1M cells. The TRA activity contained in the membrane fraction was recovered in the sodium dodecyl sulfate (SDS)-solubilized fraction after the SDS-extraction of CSA1M membranes. This CSA1M SDS-solubilized preparation gave protection against syngeneic RSV-induced CSA9F tumor cells as well as the homologous tumor cell type, but failed to induce resistance to RSV-unrelated tumor cells. The membrane or SDS-solubilized fraction from RSV-unrelated tumor cells was unable to generate anti-CSA1M protective immunity. Physicochemical analyses have demonstrated that TRA activity in the SDS-solubilized fraction was completely abolished by treatment with proteinase K but was only marginally affected after treatment with glycosidase mixture. When the SDS-solubilized preparation was applied to a Sephacryl S-300 superfine column, TRA activity was recovered in the range of molecular weight of 50-90 kD. Further fractionation of this TRA-positive fraction by SDS-polyacrylamide gel electrophoresis revealed that the molecular size of TRA is 56-68 kD. These results indicate that membrane proteins which were isolated from CSA1M tumor cells and have a molecular size of about 60 kD are capable of inducing RSV-induced tumor-specific in vivo protective immunity.
分离出了能够诱导肿瘤抗性的肿瘤抗原（肿瘤排斥抗原；TRA），并对其一些物理化学性质进行了表征。从劳氏肉瘤病毒（RSV）诱导的CSA1M肿瘤细胞中分离出胞质溶胶和质膜部分。用这些肿瘤细胞的膜部分而非胞质溶胶部分与完全弗氏佐剂一起免疫，可使小鼠对随后用活的CSA1M细胞进行的攻击产生完全保护。在对CSA1M膜进行十二烷基硫酸钠（SDS）提取后，膜部分中含有的TRA活性在SDS溶解部分中得以恢复。这种CSA1M的SDS溶解制剂能对同基因RSV诱导的CSA9F肿瘤细胞以及同源肿瘤细胞类型产生保护作用，但不能诱导对与RSV无关的肿瘤细胞的抗性。来自与RSV无关的肿瘤细胞的膜或SDS溶解部分无法产生抗CSA1M的保护性免疫。物理化学分析表明，SDS溶解部分中的TRA活性经蛋白酶K处理后完全丧失，但经糖苷酶混合物处理后仅受到轻微影响。当将SDS溶解制剂应用于Sephacryl S - 300超细柱时，TRA活性在分子量50 - 90 kD范围内得以恢复。通过SDS - 聚丙烯酰胺凝胶电泳对该TRA阳性部分进一步分级分离显示，TRA的分子大小为56 - 68 kD。这些结果表明，从CSA1M肿瘤细胞中分离出的、分子大小约为60 kD的膜蛋白能够诱导RSV诱导的肿瘤特异性体内保护性免疫。
