Yamada Shinji, Itai Shunsuke, Nakamura Takuro, Yanaka Miyuki, Kaneko Mika K, Kato Yukinari
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan.
New Industry Creation Hatchery Center, Tohoku University, 2-1, Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan.
Biochem Biophys Rep. 2018 Apr 12;14:64-68. doi: 10.1016/j.bbrep.2018.03.007. eCollection 2018 Jul.
CD44 is a transmembrane glycoprotein that regulates a variety of genes related to cell-adhesion, migration, proliferation, differentiation, and survival. A large number of alternative splicing isoforms of CD44, containing various combinations of alternative exons, have been reported. CD44 standard (CD44s), which lacks variant exons, is widely expressed on the surface of most tissues and all hematopoietic cells. In contrast, CD44 variant isoforms show tissue-specific expression patterns and have been extensively studied as both prognostic markers and therapeutic targets in cancer and other diseases. In this study, we immunized mice with CHO-K1 cell lines overexpressing CD44v3-10 to obtain novel anti-CD44 mAbs. One of the clones, CMab-5 (IgG, kappa), recognized both CD44s and CD44v3-10. CMab-5 also reacted with oral cancer cells such as Ca9-22, HO-1-u-1, SAS, HSC-2, HSC-3, and HSC-4 using flow cytometry. Moreover, immunohistochemical analysis revealed that CMab-5 detected 166/182 (91.2%) of oral cancers. These results suggest that the CMab-5 antibody may be useful for investigating the expression and function of CD44 in various cancers.
CD44是一种跨膜糖蛋白，可调节多种与细胞黏附、迁移、增殖、分化和存活相关的基因。据报道，CD44存在大量可变剪接异构体，包含可变外显子的各种组合。缺乏可变外显子的CD44标准型（CD44s）在大多数组织和所有造血细胞表面广泛表达。相比之下，CD44可变异构体表现出组织特异性表达模式，并已作为癌症和其他疾病的预后标志物和治疗靶点进行了广泛研究。在本研究中，我们用过表达CD44v3-10的CHO-K1细胞系免疫小鼠，以获得新型抗CD44单克隆抗体。其中一个克隆CMab-5（IgG，κ）可识别CD44s和CD44v3-10。使用流式细胞术，CMab-5还与口腔癌细胞如Ca9-22、HO-1-u-1、SAS、HSC-2、HSC-3和HSC-4发生反应。此外，免疫组织化学分析显示，CMab-5在182例口腔癌中检测出166例（91.2%）。这些结果表明，CMab-5抗体可能有助于研究CD44在各种癌症中的表达和功能。
