Kawaoka Toru, Oka Masaaki, Takashima Motonari, Ueno Tomio, Yamamoto Koutaro, Yahara Noboru, Yoshino Shigefumi, Hazama Shoichi
Department of Digestive Surgery and Surgical Oncology (Department of Surgery II), Yamaguchi University Graduate School of Medicine, Yamaguchi 755-8505, Japan.
Oncol Rep. 2008 Jul;20(1):155-63.
MUC1 is a tumor-associated antigen that is overexpressed in invasive ductal carcinomas of the pancreas (PC). MUC1-specific cytotoxic T lymphocytes (CTLs) recognize MUC1 molecules in a HLA-unrestricted manner. In this study, we performed adoptive immunotherapy (AIT) in patients with PC with CTLs stimulated by the MUC1-expressing human PC cell line YPK-1. To induce CTLs, peripheral blood mononuclear cells (PBMCs) were cultured for 3 days with inactivated YPK-1 cells and then stimulated with interleukin (IL)-2 for 7 days. The cytotoxicity of these cells against human cancer cell lines was analyzed, and a variety of antibodies were evaluated for their ability to inhibit cytotoxicity. We treated 8 patients with unresectable PC and 20 patients with resectable PC postsurgically. CTLs were induced as described above, suspended in 100 ml saline and injected intravenously. Induced CTLs were cytotoxic against 5 MUC1-expressing PC cell lines and a breast cancer cell line, regardless of the HLA phenotype. Low cytotoxicity was observed in 7 MUC1-negative cancer cell lines. Anti-CD3 monoclonal antibody (mAb) or anti-CD8 mAb strongly inhibited cytotoxicity against YPK-1, whereas anti-class I mAb showed no inhibition. YPK-1 cells incubated with anti-MUC1 mAb also showed low cytotoxicity. Clinically, the median survival time was 5.0 months for patients with unresectable PC treated with AIT. None of the 5 patients without liver metastasis showed hepatic recurrence. The median survival time was 17.8 months for 18 out of 20 patients with resectable PC who underwent curative surgery, and the 1-, 2- and 3-year survival rates after surgery were 83.3, 32.4, and 19.4%, respectively. Liver metastasis was found in only one patient and no side effects of AIT were observed. CTLs stimulated by a MUC1-expressing human pancreatic cancer cell line showed a strong tumor cytotoxic activity in a MUC1-specific and MHC-unrestricted manner. AIT with stimulated CTLs significantly suppressed the postsurgical hepatic recurrence of PC. Adjuvant immunotherapy with CTLs may be useful in the postsurgical treatment of PC.
黏蛋白1（MUC1）是一种肿瘤相关抗原，在胰腺浸润性导管癌（PC）中过表达。MUC1特异性细胞毒性T淋巴细胞（CTL）以不依赖人类白细胞抗原（HLA）的方式识别MUC1分子。在本研究中，我们对PC患者进行了过继性免疫治疗（AIT），所用的CTL由表达MUC1的人PC细胞系YPK-1刺激产生。为诱导CTL，外周血单个核细胞（PBMC）与灭活的YPK-1细胞共培养3天，然后用白细胞介素（IL）-2刺激7天。分析了这些细胞对人癌细胞系的细胞毒性，并评估了多种抗体抑制细胞毒性的能力。我们对8例不可切除PC患者和20例可切除PC患者术后进行了治疗。CTL按上述方法诱导产生后，悬浮于100 ml生理盐水中静脉注射。诱导产生的CTL对5种表达MUC1的PC细胞系和1种乳腺癌细胞系具有细胞毒性，与HLA表型无关。在7种MUC1阴性癌细胞系中观察到低细胞毒性。抗CD3单克隆抗体（mAb）或抗CD8 mAb强烈抑制对YPK-1的细胞毒性，而抗I类mAb未显示抑制作用。用抗MUC1 mAb孵育的YPK-1细胞也显示低细胞毒性。临床上，接受AIT治疗的不可切除PC患者的中位生存时间为5.0个月。5例无肝转移的患者均未出现肝复发。20例接受根治性手术的可切除PC患者中，18例的中位生存时间为17.8个月，术后1年、2年和3年生存率分别为83.3%、32.4%和19.4%。仅1例患者出现肝转移，未观察到AIT的副作用。由表达MUC1的人胰腺癌细胞系刺激产生的CTL以MUC1特异性和不依赖主要组织相容性复合体（MHC）的方式表现出强大的肿瘤细胞毒性活性。用刺激产生的CTL进行AIT可显著抑制PC术后肝复发。用CTL进行辅助免疫治疗可能对PC术后治疗有用。
