Ohizumi I, Tsunoda S, Taniguchi K, Saito H, Esaki K, Koizumi K, Makimoto H, Wakai Y, Matsui J, Tsutsumi Y, Nakagawa S, Utoguchi N, Ohsugi Y, Mayumi T
Department of Cancer Research, Fuji Gotemba Research Laboratories, Chugai Pharmaceutical Co. Ltd., Shizuoka, Japan.
Int J Cancer. 1998 Aug 12;77(4):561-6. doi: 10.1002/(sici)1097-0215(19980812)77:4<561::aid-ijc15>3.0.co;2-9.
We have reported the isolation and specific in vitro properties of tumor-derived endothelial cells (TEC) from rat KMT-17 fibrosarcomas transplanted into rats. To develop antibody-based tumor vascular targeting therapy for solid tumors, we have generated monoclonal antibodies (MAbs) using passive immunization of outside-out membrane vesicles of rat epididymal-fat-pad-derived capillary endothelial cells (FCEC) followed by active immunization of those of rat TEC. The MAbs produced were screened against TEC and FCEC. Of all cultured hybridomas, 75 (3.3%) of the secreted MAbs preferentially recognized TEC. We selected a total of 7 MAbs which detected antigens highly abundant in TEC, although 5 of the 7 MAbs were weakly positive for FCEC in cell-ELISA and FACS analyses. The antigens recognized by these MAbs, with the exception of MAb TES-7, were present on endothelial cells of tumor blood vessels in KMT-17 fibrosarcoma tissues, as shown by immunohistochemical analysis. Antigens of 40- and 80-kDa were recognized by MAbs TES-1, 7, 17, 21 and 26 and by MAbs TES-23 and 27 respectively. Although the function of these antigens, which are preferentially expressed on rat tumor-derived endothelial cells, is still unknown, we believe that future studies of such antigens will help elucidate the role of endothelial cells in tumor vasculature. Our results indicate that MAbs may provide a novel tool for the development of antibody-based therapy targeting tumor vasculature.
我们已报道了从移植到大鼠体内的大鼠KMT - 17纤维肉瘤中分离出的肿瘤来源内皮细胞（TEC）及其体外特异性特性。为了开发针对实体瘤的基于抗体的肿瘤血管靶向治疗方法，我们通过对大鼠附睾脂肪垫来源的毛细血管内皮细胞（FCEC）的外翻膜囊泡进行被动免疫，然后对大鼠TEC的外翻膜囊泡进行主动免疫，制备了单克隆抗体（MAb）。所产生的单克隆抗体针对TEC和FCEC进行筛选。在所有培养的杂交瘤中，75种（3.3%）分泌的单克隆抗体优先识别TEC。我们总共选择了7种单克隆抗体，它们能检测到TEC中高度丰富的抗原，尽管在细胞酶联免疫吸附测定（cell - ELISA）和荧光激活细胞分选（FACS）分析中，这7种单克隆抗体中有5种对FCEC呈弱阳性。免疫组织化学分析表明，除单克隆抗体TES - 7外，这些单克隆抗体识别的抗原存在于KMT - 17纤维肉瘤组织的肿瘤血管内皮细胞上。单克隆抗体TES - 1、7、17、21和26分别识别40 kDa和80 kDa的抗原，单克隆抗体TES - 23和27分别识别80 kDa的抗原。尽管这些优先在大鼠肿瘤来源内皮细胞上表达的抗原的功能仍然未知，但我们相信对这类抗原的进一步研究将有助于阐明内皮细胞在肿瘤血管系统中的作用。我们的结果表明，单克隆抗体可能为开发针对肿瘤血管系统的基于抗体的治疗方法提供一种新工具。
