Sato N, Yagihashi A, Okubo M, Torigoe T, Takahashi S, Sato T, Kikuchi K
Cancer Res. 1987 Jun 15;47(12):3147-51.
The molecular nature of a tumor-specific transplantation antigen (TSTA) of a chemically induced BALB/c mouse colon tumor C-C26 was investigated. The antigen was noncytolytically extracted by 2.5% n-butanol treatment of the cells. Crude butanol extract from C-C26, but not from colon tumor C-C51 and fibrosarcoma Meth-A of BALB/c mice could provide protection against the challenged C-C26 tumor in the transplantation experiment. Crude butanol extract from another syngeneic colon tumor C-C36 also induced a cross-protection against the challenged C-C26 tumor. C-C26 crude butanol extract was characterized by biochemical procedures including the Sephadex G200 column, lens culinaris affinity column, and anion-exchange Mono Q fast protein liquid chromatography column, and by the enzyme digestion study of the antigens and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The data indicated that C-C26 TSTA was eluted into fractions containing molecules of approximately Mr 200,000 on Sephadex G200 column chromatography. This antigen was also found in unbound fractions on a lens culinaris affinity column. The antigen was further separated into the fraction that was eluted with 0.4 M NaCl in an ionic strength on Mono Q fast protein liquid chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of this fraction showed the molecule with a molecular weight of 30,000. The enzyme digestion study indicated that the immunogenicity of the antigen was inactivated by papain but probably not by neuraminidase treatment. These data suggest that the immunogenic moiety of C-C26 TSTA molecules is located in the peptide portions rather than in sialic acid residues or carbohydrate portions. Furthermore, there are several similarities of the molecular characteristics between C-C26 TSTA and previously reported C-C36 TSTA, such as the amenability to n-butanol extraction. Lens culinaris lectin inaffinity, and ionic strength.
对化学诱导的BALB/c小鼠结肠肿瘤C-C26的肿瘤特异性移植抗原（TSTA）的分子性质进行了研究。通过用2.5%正丁醇处理细胞以非细胞溶解方式提取该抗原。在移植实验中，来自C-C26的粗正丁醇提取物，而非来自BALB/c小鼠的结肠肿瘤C-C51和纤维肉瘤Meth-A的粗正丁醇提取物，能够提供针对受攻击的C-C26肿瘤的保护作用。来自另一种同基因结肠肿瘤C-C36的粗正丁醇提取物也诱导了针对受攻击的C-C26肿瘤的交叉保护作用。通过包括Sephadex G200柱、刀豆球蛋白A亲和柱和阴离子交换Mono Q快速蛋白质液相色谱柱在内的生化方法，以及对抗原的酶消化研究和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，对C-C26粗正丁醇提取物进行了表征。数据表明，在Sephadex G200柱色谱上，C-C26 TSTA被洗脱到含有分子量约为200,000的分子的级分中。在刀豆球蛋白A亲和柱上的未结合级分中也发现了这种抗原。在Mono Q快速蛋白质液相色谱上，该抗原进一步被分离到在离子强度为0.4M NaCl时洗脱的级分中。该级分的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示分子量为30,000的分子。酶消化研究表明，木瓜蛋白酶可使抗原的免疫原性失活，但神经氨酸酶处理可能不会使其失活。这些数据表明，C-C26 TSTA分子的免疫原性部分位于肽段，而非唾液酸残基或碳水化合物部分。此外，C-C26 TSTA与先前报道的C-C36 TSTA在分子特征上有几个相似之处，如对正丁醇提取的适应性、刀豆球蛋白A凝集素不亲和性和离子强度。
