New immuno Adoptive tangeting thesapy using bispecific Amtibody

为了建立一种新的胆管癌过继免疫治疗方法，我们合成了两种双特异性抗体（BsAbs）：MUC 1 * CD 3 BsAb和MUC 1 * CD 28 BsAb。MUC 1 * CD 3 BsAb由MUE 11（抗MUC 1肿瘤抗原）和OKT-3（抗CD 3）组成，MUC 1 * CD 28 BsAb由MUE 11和15 E8（抗CD 28）组成。这两种BsAb与MUC 1阳性的靶肿瘤细胞和效应淋巴因子激活的杀伤（LAK）细胞反应良好。体外细胞毒试验[3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四氮唑溴化物法]显示，MCU 1 * CD 3 BsAb能抗原特异性地增强LAK细胞的细胞毒活性。两种BsAbS（MUC 1 * CD 3 BsAb加MUC 1 * CD 28 BsAb）的体外加入导致60%的细胞毒性，与单独使用BsAb（MUC 1 * CD 3）获得的细胞毒性相似。白细胞介素12诱导的LAK细胞表现出更大的细胞毒性（50%）比他们的白细胞介素2诱导的同行（LAK细胞），这也增强了BsAb。当用两种BsAb致敏的2*10^7个LAK细胞给药四次时，观察到肿瘤生长的抑制。i.v.to因此，BsAb-LAK治疗控制BDC保证临床试验。

For the purpose of establishing a new adoptive immunotherapy for bile duct carcinoma (BDC), we synthesized two bispecific antibodies (BsAbs), MUC1*CD3 BsAb constructed with MUSE11 (anti-MUC1 tumor antigen) and OKT-3 (anti-CD3), and MUC1*CD28 BsAb constructed with MSUE11 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 MCU1*CD3 BsAb could antigen-specifically enhance the cytotoxicity of LAK cells. Addition of the two BsAbS (MUC1*CD3 BsAb plus MUC1*CD28 BsAb) in vitro resulted in a 60% cytotoxicity, similar to that obtained with BsAb (MUC1*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*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.