ISOLATION AND CHARACTERIZATION OF A MONOCLONAL-ANTIBODY, K1, REACTIVE WITH OVARIAN CANCERS AND NORMAL MESOTHELIUM
ISOLATION AND CHARACTERIZATION OF A MONOCLONAL-ANTIBODY, K1, REACTIVE WITH OVARIAN CANCERS AND NORMAL MESOTHELIUM
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DOI：
10.1002/ijc.2910500308
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发表时间：
1992-02-01
影响因子：
6.4
通讯作者：
WILLINGHAM, MC
中科院分区：
文献类型：
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作者：
CHANG, K;PASTAN, I;WILLINGHAM, MC
我们已经分离到一种新的单克隆抗体（MAb），K1，它与人卵巢癌细胞表面的表位反应。该抗体通过用高碘酸盐处理的人卵巢癌（OVCAR-3）细胞免疫小鼠产生。这些小鼠先前已经对正常的人类肾膜产生了耐受性。在融合之前，使用淘选纯化方法在活OVCAR-3细胞上选择来自这些小鼠的脾淋巴细胞。在一天内使用免疫荧光对活OVCAR-3细胞进行表面反应性克隆的初步筛选，并使用免疫过氧化物酶组织化学对正常人组织和人肿瘤的冷冻切片进行二次筛选。将K1克隆亚克隆并鉴定为IgM同种型，但随后使用淘选选择方法将同种型转换为IgG 1 K。当通过免疫组化进行评价时，在许多卵巢非粘液性肿瘤以及食管鳞状肿瘤和宫颈癌中发现了与K1反应的抗原。与K1显示一致反应性的唯一正常成人组织是腹膜、胸膜和心包腔的间皮膜。与气管、扁桃体和输卵管上皮细胞的反应也有限。在食蟹猴组织中发现了与K1相似的组织反应性，K1与许多与先前鉴定的抗体OC 125相同的组织和肿瘤反应，但几条证据表明，与OC 125相比，K1与不同的表位反应，可能是不同的分子。这一证据包括采用免疫荧光竞争，双标记免疫荧光，固相和活细胞放射免疫分析。由于我们的数据表明与K1抗体反应的抗原是一种新的分子种类，我们将抗原命名为CAK 1。与脱落抗原CA 125不同，CAK 1仅与细胞相关，并且在培养的OVCAR-3细胞的上清液或卵巢癌患者的血液中未发现。K1抗体可用作治疗和诊断卵巢癌以及一些其它人类癌症的靶向剂。
We have isolated a new monoclonal antibody (MAb), K1, that reacts with an epitope on the surface of human ovarian carcinoma cells. This antibody was generated by immunization of mice with periodate-treated human ovarian carcinoma (OVCAR-3) cells. These mice had been previously made tolerant with normal human kidney membranes. Spleen lymphocytes from these mice were selected prior to fusion using a panning purification method on living OVCAR-3 cells. Initial screening of surface-reactive clones was performed in a single day using immunofluorescence on living OVCAR-3 cells, and secondary screening was performed using immunoperoxidase histochemistry on cryostat sections of normal human tissues and human tumors. The K1 clone was subcloned and identified as an IgM isotype, but was subsequently isotype-switched to IgG1K using a panning selection method. When evaluated by immunohistochemistry, the antigen reactive with K1 was found in many ovarian non-mucinous tumors, as well as in squamous tumors of the esophagus, and cervical cancer. The only normal adult human tissues showing uniform reactivity with K1 were the mesothelia of the peritoneal, pleural and pericardial cavities. There was also limited reactivity with epithelia of the trachea, tonsil and Fallopian tube. A similar tissue reactivity for K1 was found in tissues from cynomolgus monkeys, K1 reacted with many of the same tissues and tumors as the previously identified antibody OC125, but several lines of evidence indicate that K1 reacts with a different epitope and probably a different molecule, when compared to OC125. This evidence included assays employing immunofluorescence competition, double-label immunofluorescence, and solid-phase and live-cell radioimmunoassays. Since our data indicate that the antigen reactive with the K1 antibody is a new molecular species, we have named the antigen CAK1. Unlike the shed antigen CA125, CAK1 was only cell-associated and was not found in the supernatant of cultured OVCAR-3 cells or in the blood of ovarian cancer patients. The K1 antibody may be useful as a targeting agent for therapy and in the diagnosis of ovarian carcinoma, as well as some other human cancers.