Expression of MAGE tumor rejection antigen on lung cancer and application for cancer vaccine

(1)用GST-MAGE-1或-4融合蛋白刺激肺癌患者外周血单个核细胞（PBMC）。但不能诱导出针对法师抗原的特异性CTL。另一方面，通过在体外用限制在HLA-A2中的MART-1肽刺激，可以诱导针对该肽的特异性CTL（Sugita et al.，国际免疫学，8（5），1996）。(2)法师基因经常在许多不同的癌症中表达，但在除睾丸和胎盘之外的正常细胞或正常组织中不表达。法师家族蛋白的生物学功能有待进一步研究。用抗重组法师-1或法师-4蛋白的抗体，用免疫组化方法研究了法师-1或法师-4蛋白在睾丸细胞中的表达。这两种蛋白质被确定在精原细胞和初级精母细胞的细胞核和细胞质中，但在精子细胞或Sertoli细胞中没有。因此，法师蛋白是正常组织的抗原区室， ...更多信息 特别是在精子发生的早期阶段起重要作用的睾丸细胞（Takahashi等，Cancer Res.，55，1995）。(3)使用针对重组法师-4 b蛋白的单克隆抗体和多克隆抗体建立酶联免疫吸附测定（ELISA）来测量在人肿瘤细胞上表达的细胞法师-4蛋白（Shichijo et al.，J Immunol. Method，186，1995）。(4)我们研究了原发性肺癌患者血清中法师-4蛋白的水平，以了解法师蛋白的生物学作用。在法师-4^+肿瘤细胞系的上清液和患者的血清中检测到法师-4蛋白是未降解的。肺癌患者血清法师-4蛋白水平（n=100，平均1.17 ng/ml）显著高于肺良性疾病患者（n=80，033 ng/ml）和健康献血员（n=68，032 ng/ml）。100名癌症患者中有34名高于临界水平（1.15ng/ml），但其他组中没有任何人高于临界水平（Shichijo et al.，JJCR，in press，1997）. (5)本文对16例非小细胞肺癌患者IL-2激活的肿瘤浸润淋巴细胞进行了HLA Ⅰ类限制性和肿瘤特异性细胞毒活性的研究。建立了六个HLA I类限制性和肿瘤特异性CTL系：（i）三名腺癌患者中的HLA-A2限制性和腺癌特异性CTL，（ii）HLA-A3101/3302+腺癌患者中的HLA-A3101-和A3302限制性和腺癌特异性CTL，和（iii）识别HLA-A3302+腺癌患者和HLA-A2402鳞状细胞癌患者中具有不同组织学类型的肿瘤的HLA-A3302限制性CTL和（iv）识别HLA-A2402限制性CTL，分别（Seki等人，Cell.免疫学，175，1997）。这些结果表明，存在HLA I类限制性和肿瘤特异性的CTL识别腺癌或SCC的HLA-A等位基因的肽抗原，

(1) Peripheral blood mononuclear cells (PBMC) from lung cancer patients had been stimulared with GST-MAGE-1 or-4 fusion protein. However, we could not induct specific CTL against MAGE antigens. In the other hand, it had been possible to induce specific CTLs against MART-1 peptide restricted in HLA-A2 by the stimulation with this peptide in vitro (Sugita et al., Int. Immunol., 8 (5), 1996). (2) The MAGE genes are frequently expressed in many different cancers, but are not expressed in normal cells or normal tissues other than testis and placenta. Biological roles of proteins of MAGE family remained to be investigated. The expression of MAGE-1 or MAGE-4 protein in the testicular cells was studied immunohistochemically with the antibodies to the recombinant MAGE-1 or MAGE-4 protein. Both proteins were identified in the nucleus and cytoplasm of spermatogonia and in primary spermatocytes but not in spermatids or Sertoli's cells. Therefore, MAGE proteins are normal tissue antigens compartmen … More talized in particular testicular cells playing an important role in the early phase of the spermatogenesis (Takahashi et al, Cancer Res., 55,1995). (3) An enzyme-linked immunosorbent assay (ELISA) was established for measuring cellular MAGE-4 protein expressed on human tumor cells using a monoclonal antibody and polyclonal Ab to recombinant MAGE-4b protein (Shichijo et al., J Immunol. Method, 186,1995). (4) We investigated the level of MAGE-4 protein in sera of patients with primary lung cancer to understand the biological roles of the MAGE proteins. MAGE-4 protein was detected as a non-degraded from in both the supernatant of a MAGE-4^+ tumor cell line and in a patient's serum. Surum level of the MAGE-4 protein in lung cancer patients (n=100, mean=1.17ng/ml) was significantly higher than that in either patients with benign pulmonary diseases (n=80,033ng/ml) or healthy donors (n=68,032ng/ml). It was higher than the cutoff level (1.15ng/ml) in 34 of 100 cancer patients, but not in anyone in the other groups (Shichijo et al., JJCR,in press, 1997). (5) HLA class lrestriction and tumor specificity of cytotoxicity in the IL-2-activated tumor-infiltrating lymphocytes from 16 patients with non-small cell lung cancer were investigated. Six HLA class l-restricted and tumor-specific CTL lines were established : (i) HLA A2-restricted and adenocarcinoma-specific CTLs in three patients with adenocarcinoma, (ii) HLA-A3101-and A3302-restricted and adenocarcinoma-specific CTLs in an HLA-A3101/3302+patients with adenocarcinoma, and (iii) HLA-A3302-restricted CTLs and (iv) HLA-A2402-restricted CTLs recognizing tumors with different types of histology in an HLA-A3302+patients with adenocarcinoma and HLA-A2402 patients with squamous cell carcinoma, respectively (Seki et al., Cell. Immunol., 175,1997). These results showed evidence of the existence of HLA class l-restricted and tumor-specific CTLs recognizing peptide antigens on HLA-A alleles of adenocarcinoma or SCC in tumor sites of a Less