Yoshikawa K, Furukawa K, Ueda R, Iwasa S, Lloyd K O, Notake K, Takahashi T
Department of Microbiology, Aichi Medical University, Japan.
Jpn J Cancer Res. 1989 Jun;80(6):546-53. doi: 10.1111/j.1349-7006.1989.tb01674.x.
Lymph-node lymphocytes of a patient with stomach cancer were fused with the mouse-human heterohybridoma, HM-5. A clone (2F9) was isolated that showed stable production of an IgM antibody reactive with NUGC-4 stomach cancer cell line. This antibody reacted predominantly with a cell surface antigen on cell lines originating from gastro-intestinal cancer and adenocarcinoma of lung, whereas it was not generally reactive with other types of cancers, or with normal kidney cells or fibroblasts. Biotin-labeled 2F9 antibody clearly stained cell smears and the nude mouse tumor of NUGC-4, but it did not show a positive reaction with stomach cancer tissues obtained from more than 10 patients, indicating that the antigen detected is very weakly expressed on tumor cells or on a limited number of stomach cancers. The antigen shed from NUGC-4 cell line was detected in the culture supernatant. 2F9 antibody precipitated a glycoprotein with a molecular weight of over 200 kilodaltons as well as a possible glycolipid, from NUGC-4 cells labeled with [3H]glucosamine or [35S]-H2SO4. Periodic acid treatment of the tissue section decreased reactivity with 2F9 antibody, but heat, neuraminidase or protease treatment did not. These results suggested that the epitope is present on a carbohydrate moiety not containing sialic acid, and that a part of the antigen molecule is sulfated.
将一名胃癌患者的淋巴结淋巴细胞与小鼠-人异源杂交瘤HM-5进行融合。分离出一个克隆（2F9），该克隆显示能稳定产生与NUGC-4胃癌细胞系反应的IgM抗体。这种抗体主要与源自胃肠道癌和肺腺癌的细胞系上的细胞表面抗原发生反应，而一般不与其他类型的癌症、正常肾细胞或成纤维细胞发生反应。生物素标记的2F9抗体能清晰地对NUGC-4的细胞涂片和裸鼠肿瘤进行染色，但对10多名患者的胃癌组织未显示阳性反应，这表明所检测到的抗原在肿瘤细胞上表达非常弱，或仅在少数胃癌上表达。在NUGC-4细胞系的培养上清液中检测到了从该细胞系脱落的抗原。2F9抗体从用[3H]葡糖胺或[35S]-H2SO4标记的NUGC-4细胞中沉淀出一种分子量超过200千道尔顿的糖蛋白以及一种可能的糖脂。对组织切片进行高碘酸处理会降低其与2F9抗体的反应性，但加热、神经氨酸酶或蛋白酶处理则不会。这些结果表明，表位存在于不含唾液酸的碳水化合物部分，且抗原分子的一部分被硫酸化。
