Watanabe M, Hirohashi S, Shimosato Y, Ino Y, Yamada T, Teshima S, Sekine T, Abe O
Jpn J Cancer Res. 1985 Jan;76(1):43-52.
A monoclonal antibody, ST-4-39, was obtained by using a human gastric cancer xenograft, St-4, as an immunogen. Immunization was achieved by transferring immunocompetent mouse spleen cells into a nude mouse bearing St-4. Hybridomas were produced with the spleen cells of the mouse after rejection of the tumor and screened for immunohistochemical reactivity with cancers and normal tissues on formalin-fixed paraffin sections. ST-4-39 immunohistochemically reacted with various cancers including gastric, colorectal and pancreatic cancers as well as some normal tissues. ST-4-39 and NS 19-9 differed in immunohistochemical reactivity, although they reacted with some cancers and a few normal tissues in common. PBS extracts of normal and cancer tissues were examined for antigen reactive with ST-4-39 by sandwich enzyme immunoassay. Extractable antigen was detected in adenocarcinomas of colon, stomach and lung, while it was detected only in salivary gland and trachea among normal tissues examined. Gel filtration analysis of the antigen indicated a molecular weight of greater than or equal to 1 X 10(6), and the antigenic determinant was suggested to be a carbohydrate chain with terminal sialic acid by studies using periodic acid, neuraminidase and pronase treatments. Furthermore, the ST-4-39 antigen affinity-purified from two gastric cancer strains was shown to contain multiple carbohydrate determinants including sialyl-Lewisa and sialyl-Lewisx, suggesting the antigen to be a mucin.
以人胃癌移植瘤St-4作为免疫原获得了一种单克隆抗体ST-4-39。通过将具有免疫活性的小鼠脾细胞移植到携带St-4的裸鼠中来实现免疫。在肿瘤被排斥后,用小鼠的脾细胞制备杂交瘤,并在福尔马林固定石蜡切片上筛选与癌组织和正常组织的免疫组织化学反应性。ST-4-39与包括胃癌、结直肠癌和胰腺癌在内的各种癌症以及一些正常组织发生免疫组织化学反应。ST-4-39和NS 19-9在免疫组织化学反应性上有所不同,尽管它们与一些癌症和少数正常组织有共同反应。通过夹心酶免疫测定法检测正常组织和癌组织的PBS提取物中与ST-4-39反应的抗原。在结肠、胃和肺的腺癌中检测到可提取抗原,而在所检测的正常组织中仅在唾液腺和气管中检测到。对抗原的凝胶过滤分析表明其分子量大于或等于1×10(6),并且通过使用高碘酸、神经氨酸酶和链霉蛋白酶处理的研究表明抗原决定簇是带有末端唾液酸的碳水化合物链。此外,从两种胃癌菌株中亲和纯化的ST-4-39抗原显示含有多种碳水化合物决定簇,包括唾液酸化路易斯a和唾液酸化路易斯x,提示该抗原为粘蛋白。