Establishment of tumor specific killer T-cells and the cloning of the tumor rejection antigen gene

我们研究了食管癌肿瘤部位HLA I类限制性和肿瘤特异性CTL的存在。从15名患者中的5名的转移位点建立HLA I类限制性和肿瘤特异性CTL系（Toh et.例如，Cell. 177：137- 143，1997）。与此相反，在4例转移性LN患者中，接受了术前化疗和/或放疗，或在6例原发性食管肿瘤中，均无TIL增殖。5例患者非转移区淋巴细胞均未产生CTL。这些结果表明，在未经术前化疗和/或放疗的食管鳞状细胞癌患者的肿瘤部位存在HLA-I类限制性和肿瘤特异性CTL。我们从食管鳞状细胞癌（SCC）的cDNA文库中鉴定了编码肿瘤抗原的基因，该基因可被HLA-A2601限制性CTL识别（Shichijo等人，J.Exp. 187：277- 288，1998）。该基因与已知序列没有相似性，编码两种蛋白质（125和43 kD）。125 kD的蛋白质与亮氨酸拉链基序表达在大多数增殖细胞的核测试，包括正常和恶性细胞。43 kD蛋白表达于大多数来自不同器官的SCC和一半的肺腺癌的胞浆中，但在其他癌症和一组正常组织中不表达。两种蛋白共有的三个九肽被KE 4 CTL识别，并且其中一个肽在体外从PBMC诱导的CTL仅限于自体肿瘤细胞。43 kD蛋白和该九肽（KGSGKMKTE）可能用于HLA-A2601^+上皮癌患者的特异性免疫治疗。

We in vestigated the presence of HLA-class I-restricted and tumor-specific CTL in tumor sites of esophageal can cers. HLA-class I-restricted and tumor-specific CTL lines were established from metastatic sites in 5 of 15 patients (Toh et. al., Cell. Immunol. 177 : 137-143,1997). In contrast to these five cases, there was no TIL proliferation in any of four cases of metastatic LN that had undergone prepperative chemotherapy and/or radiation or in six cases of original esophageal tumors. CTL were not generated from lymphocytes of non-metastatic region al LN in any of five patients. These results suggest the existence of HLA-class I-restricted and tumor-specific CTL in tumor sites of esophageal SCC patients who had no preoperative chemotherapy and/or radiation.We have identified a gene encoding tumor antigens from cDNA library of esophageal squamous cell carcinomas (SCCs) recognizes by HLA-A2601-restricted CTLs (Shichijo et al., J.Exp. Med. 187 : 277-288,1998). This gene showed no similarity to known sequences, and encoded two (125 and 43 kD) proteins. The 125 kD protein with leucine-zipper motif was expressed in the nucleus of the majority of proliferating cells tested including normal and malignant cells. The 43 kD protein was expressed in the cytosol of most of SCCs from various organs and half of lung adenocarcinomas, but was not expressed in other cancers nor in a panel of normal tissues. The three nonapeptides in the region shared by the two proteins were recognized by the KE4 CTLs, and one of the peptides induced in vitro from PBMCs the CTLs restricted to the autologous tumor cells. The 43 kD protein and this nonapeptide (KGSGKMKTE) may be useful for specific immunotherapy of HLA-A2601^+ epithelial cancer patients.