Ohuchi N, Wunderlich D, Fujita J, Colcher D, Muraro R, Nose M, Schlom J
Cancer Res. 1987 Jul 1;47(13):3565-71.
Murine monoclonal antibodies (MAbs) reactive with distinct epitopes on carcinoembryonic antigen (CEA) have been analyzed systematically by radioimmunoassays, Western blotting, and immunohistochemical assays to define CEA expression in adenocarcinomas, benign lesions, and normal tissues of the stomach. Each of four COL-MAbs (COL-1, COL-4, COL-6, and COL-12) reacted preferentially with cell extracts of adenocarcinomas versus those of normal mucosae in solid-phase radioimmunoassays. Using Western blotting analyses MAbs COL-1, COL-4, COL-6, and COL-12 detected only the Mr 180,000 molecule characteristic of CEA in adenocarcinoma of the stomach; no reactivity was observed in an extract of normal gastric mucosa. Antibody competition radioimmunoassays were then carried out to define relations among COL-MAbs using 125I-radiolabeled MAbs, and nonradiolabeled MAbs as competitors. A spectrum of formalin-fixed, paraffin-embedded normal, benign, and malignant tissue sections of the stomach were examined for immunoreactivities with COL-MAbs using immunohistochemical assays to define whether the COL-MAbs were able to detect CEA expression in early foci of gastric carcinomas. All of the COL-MAbs generally demonstrated selective reactivities to adenocarcinomas (n = 40) versus benign lesions (n = 15) and normal mucosae (n = 6) of the stomach. From 72 to 100% of adenocarcinomas at early stage (n = 18) were reactive with the COL-MAbs, suggesting that these MAbs might serve as immunohistochemical diagnostic tools to detect early foci of gastric carcinoma. The data reported here indicate that the COL-MAbs can potentially be utilized as radioimmunological and immunohistochemical adjuncts to differentiate early adenocarcinomas from normal mucosae or benign lesions of the stomach on the basis of differential CEA expression.
已通过放射免疫测定、蛋白质印迹法和免疫组织化学测定系统分析了与癌胚抗原(CEA)上不同表位发生反应的鼠单克隆抗体(MAb),以确定CEA在胃腺癌、良性病变和正常组织中的表达情况。在固相放射免疫测定中,四种COL-MAb(COL-1、COL-4、COL-6和COL-12)中的每一种与腺癌细胞提取物的反应均优先于与正常黏膜细胞提取物的反应。使用蛋白质印迹分析,MAb COL-1、COL-4、COL-6和COL-12仅在胃腺癌中检测到了CEA特有的180,000道尔顿分子;在正常胃黏膜提取物中未观察到反应性。然后进行抗体竞争放射免疫测定,以使用125I放射性标记的MAb和未放射性标记的MAb作为竞争者来确定COL-MAb之间的关系。使用免疫组织化学测定检查了一系列福尔马林固定、石蜡包埋的胃正常、良性和恶性组织切片与COL-MAb的免疫反应性,以确定COL-MAb是否能够检测胃癌早期病灶中的CEA表达。所有COL-MAb通常对胃腺癌(n = 40)与良性病变(n = 15)和正常黏膜(n = 6)表现出选择性反应性。72%至100%的早期腺癌(n = 18)与COL-MAb发生反应,这表明这些MAb可能用作检测胃癌早期病灶的免疫组织化学诊断工具。此处报告的数据表明,基于CEA表达差异,COL-MAb有潜力用作放射免疫和免疫组织化学辅助手段,以区分胃早期腺癌与正常黏膜或良性病变。