Katayose Y, Kudo T, Suzuki M, Shinoda M, Saijyo S, Sakurai N, Saeki H, Fukuhara K, Imai K, Matsuno S
First Department of Surgery, Tohoku University School of Medicine, Sendai, Japan.
Cancer Res. 1996 Sep 15;56(18):4205-12.
For the purpose of establishing a new adoptive immunotherapy for bile duct carcinoma (BDC), we synthesized two bispecific antibodies (BsAbs), MUC1 x CD3 BsAb constructed with MUSE11 (anti-MUC1 tumor antigen) and OKT-3 (anti-CD3), and MUC1 x CD28 BsAb constructed with MUSE11 and 15E8 (anti-CD28) antibodies. These two BsAbs reacted well with both MUC1-positive target tumor cells and effector lymphokine-activated killer (LAK) cells. Investigation of in vitro cytotoxicity [3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyltetrazolium bromide assay] revealed that the MUC1 x CD3 BsAb could antigen-specifically enhance the cytotoxicity of LAK cells. Addition of the two BsAbs (MUC1 x CD3 BsAb plus MUC1 x CD28 BsAb) in vitro resulted in a 60% cytotoxicity, similar to that obtained with BsAb (MUC1 x CD3) alone. Interleukin 12-induced LAK cells demonstrated far greater cytotoxicity (50%) than their interleukin 2-induced counterparts (LAK cells), and this was also enhanced by the BsAbs. When 2 x 10(7) LAK cells sensitized with both kinds of BsAbs were administered four times i.v. to BDC-grafted severe combined immunodeficient mice (tumor size 5 mm in diameter), inhibition of tumor growth was observed. Thus, BsAb-LAK therapy for control of BDC warrants clinical trials.
为建立一种新的胆管癌（BDC）过继性免疫疗法，我们合成了两种双特异性抗体（BsAbs），即由MUSE11（抗MUC1肿瘤抗原）和OKT - 3（抗CD3）构建的MUC1 x CD3 BsAb，以及由MUSE11和15E8（抗CD28）抗体构建的MUC1 x CD28 BsAb。这两种BsAbs与MUC1阳性靶肿瘤细胞和效应淋巴细胞激活的杀伤细胞（LAK）均反应良好。体外细胞毒性研究[3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐检测]显示，MUC1 x CD3 BsAb可抗原特异性增强LAK细胞的细胞毒性。体外添加这两种BsAbs（MUC1 x CD3 BsAb加MUC1 x CD28 BsAb）导致60%的细胞毒性，与单独使用BsAb（MUC1 x CD3）时相似。白细胞介素12诱导的LAK细胞表现出比白细胞介素2诱导的LAK细胞更强的细胞毒性（50%），并且这种细胞毒性也被BsAbs增强。当用两种BsAbs致敏的2×10⁷LAK细胞经静脉注射四次给予BDC移植的严重联合免疫缺陷小鼠（肿瘤直径5 mm）时，观察到肿瘤生长受到抑制。因此，BsAb - LAK疗法用于控制BDC值得进行临床试验。
