Ohyanagi H, Ishida H, Ishida T, Soyama N, Yamamoto M, Okumura S, Kano Y, Ueda Y, Saitoh Y
First Department of Surgery, Kobe University School of Medicine.
Jpn J Cancer Res. 1988 Dec;79(12):1349-58. doi: 10.1111/j.1349-7006.1988.tb01566.x.
A monoclonal antibody, KM10 (IgG1) was produced by fusing spleen cells from a human gastric cancer cell (MKN45)-primed BALB/c mouse with the murine myeloma cell line X63-Ag8-653. The antibody reacted strongly with the plasma membrane of human gastrointestinal carcinoma. Sections of the malignant and benign tissues were tested with immunoperoxidase. All of 10 (100%) large intestinal cancers, 26 of 31 (84%) gastric cancers, 5 of 7 (71%) pancreatic cancers and all of 3 (100%) ampullary cancers reacted positively. Moderate or weak reactivity was observed with normal human tissues, hepatoma and carcinomas of mammary, thyroid and adrenal glands. According to a study of the distribution of 125I-labeled KM10 in nude mice bearing human gastric cancer, KM10 selectively localized in tumor tissue rather than normal tissue. Whole body autoradiography also supported such a selective distribution. Destruction of antigenic properties by pronase digestion demonstrated its protein nature and by Western blot analysis, it was identified as a protein with an Mr of 180-200 kd. KM10-adriamycin (ADM) conjugate was prepared via an oxidized dextran bridge and this immunoconjugate retained the binding activity against human gastric cancer. MKN45 cells were inoculated subcutaneously into athymic mice and intravenous treatment was begun when the tumor became measurable. A dose-dependent antitumor activity was observed in vivo with KM10-ADM conjugate, while this conjugate was less toxic than free ADM.
通过将来自用人类胃癌细胞（MKN45）免疫的BALB/c小鼠的脾细胞与鼠骨髓瘤细胞系X63-Ag8-653融合，制备了单克隆抗体KM10（IgG1）。该抗体与人胃肠道癌的质膜强烈反应。用免疫过氧化物酶检测恶性和良性组织切片。10例（100%）大肠癌、31例中的26例（84%）胃癌、7例中的5例（71%）胰腺癌以及3例（100%）壶腹癌均呈阳性反应。在正常人体组织、肝癌以及乳腺、甲状腺和肾上腺的癌组织中观察到中度或弱反应性。根据对125I标记的KM10在荷人胃癌裸鼠体内分布的研究，KM10选择性地定位于肿瘤组织而非正常组织。全身放射自显影也支持这种选择性分布。链霉蛋白酶消化破坏抗原特性证明了其蛋白质性质，通过蛋白质印迹分析，它被鉴定为一种分子量为180 - 200kd的蛋白质。通过氧化葡聚糖桥制备了KM10 - 阿霉素（ADM）偶联物，该免疫偶联物保留了对人胃癌的结合活性。将MKN45细胞皮下接种到无胸腺小鼠体内，当肿瘤可测量时开始静脉内治疗。在体内观察到KM10 - ADM偶联物具有剂量依赖性抗肿瘤活性，而该偶联物的毒性比游离阿霉素小。
