Development of cancer vaccine with MAGE gene products.

MAGE-1、-2、-3、-4、-6和-12基因经常在许多不同的癌症中表达，但在睾丸和胎盘以外的正常细胞或正常组织中不表达。MAGE-1和-3肽目前被用作癌症患者的疫苗，12例HLA-A1转移性黑色素瘤患者中有3例对MAGE-3肽有反应。因此，MAGE蛋白可能成为HLA-A1癌症患者的潜在疫苗。然而，在应答患者的外周血中未检测到CTL，并且没有可用的监测方法来了解MAGE疫苗的疗效。此外，MAGE家族蛋白的生物学作用仍有待研究。我们研究了不同癌症患者血清MAGE-4蛋白（MAGE蛋白家族中的一员）的水平。在不同类型肿瘤（肺癌、头颈癌和肝细胞癌）患者的MAGE-4 +细胞系上清和血清中均检测到MAGE-4蛋白的非降解形式。例如，肺癌患者（n=100，平均=1.17ng/ml）的血清MAGE-4蛋白水平显著（p=0.0013）高于良性肺部疾病患者（n=80，平均=0.33ng/ml）或HD患者（n=68，平均=0.32ng/ml） （Shichijo et al.， JJCR,in press, 1977）。100例肿瘤患者中有34例血清MAGE-4水平高于临界值（1.15ng/ml），其余组均未达到临界值。在其他癌症患者的血清中也得到了类似的结果。此外，值得注意的是，手术切除肿瘤块可导致血清MAGE-4蛋白水平降低，复发与血清MAGE-4蛋白水平升高相关（Iwamoto et al., Int.）。j .癌症出版社，1997)。此外，在肝细胞癌和丙型肝炎病毒阳性肝硬化（肝细胞癌的高危组）患者中，血清MAGE-4水平均显著升高。这些信息对于更好地理解MAGE蛋白的生物学作用可能是重要的，并且测量MAGE-4蛋白的血清水平可能有助于检测具有不同组织学类型的MAGE-4^+癌症。在自体肿瘤细胞或il -2激活的肿瘤浸润淋巴细胞（til）刺激的黑素瘤患者外周血单个核细胞（PBMC）的T细胞中观察到HLA - i类限制性和肿瘤特异性ctl。利用这些ctl从黑色素瘤中克隆出了编码被ctl识别的肽抗原的基因。然而，这些ctl或肽抗原的存在在腺癌或鳞状细胞癌中都很少有报道，这两种主要的人类癌症需要开发新的治疗方式。我们研究了食管癌、胃癌、结肠癌和肺癌中IL-2激活的til的HLA-A位点限制和肿瘤特异性。结果显示，食管癌TILs中存在HLA类限制性和肿瘤特异性的ctl (Toh等，Cell immune。in press, 1997)，胃癌(Hoshino et al., Int。J.Cancer in press, 1997)，结肠癌（Gouhara et al.， JJCR,in press, 1997）和非小细胞肺癌（Seki et al., Cell immunolo）。In press, 1997)。这些ctl可能是鉴定编码肿瘤排斥抗原的基因的工具，用于开发癌症疫苗。少

The MAGE-1, -2, -3, -4, -6, and -12genes are frequently expressed in many different cancers, but are not expressed in normal cells or normal tissues other than testis and placenta. MAGE-1 and -3 peptides are currently used as vaccines for cancer patients, and three of 12 HLA-A1 patients with metastatic melanoma responded to MAGE-3 peptide. Therefore, MAGE protein could be a potential vaccine for HLA-A1 cancer patients. However, CTL were not detected in the peripheral blood of the responding patients, and there was no available monitoring methods to know the efficacy of MAGE vaccine. Further, biological roles of proteins of MAGE family remain to be investigated. We investigated serum level of MAGE-4 protein, one of the family of MAGE proteins, in various cancer patients. MAGE-4 protein was detected as a non-degraded form in both the supernatant of MAGE-4^+ tumor cell line and serum of cancer patients with different types of histology (lung cancer, head and neck cancer, and hepatocellula … More r carcinomas. For example, serum level of the MAGE-4 protein of lung cancer patients (n=100, mean=1.17ng/ml) was significantly (p=0.0013) higher than that of either patients with benign pulmonary diseases (n=80, mean=0.33ng/ml) or HD (n=68, mean=0.32ng/ml) (Shichijo et al., JJCR,in press, 1977). The serum level of MAGE-4 was higher than the cutoff level (1.15ng/ml) in 34 of 100 cancer patients, but no one in the other groups reached the cutoff level. The similar results were obtained in sera of the other cancer patients. In addition, it is of note that surgical removal of tumor mass resulted in decrease of serum level of MAGE-4 protein and recurrence was associated with it's reincrease (Iwamoto et al., Int. J.Cancer, in press, 1997). Furthermore, serum MAGE-4 was significantly higher in patients with both hepatocellular carcinoma and hepatitis C virus positive liver cirrhosis, a high risk group of hepatocellular carcinoma. These information could be important for better understanding of biological roles of MAGE proteins, and measurement of serum levels of MAGE-4 protein could be useful for detection of MAGE-4^+ cancers with different types of histology.HLA class I-restricted and tumor-specific CTLs have been observed in T cells of peripheral blood mononuclear cells (PBMC) stimulated with autologous tumor cells or IL-2-activated tumor infiltrating lymphocytes (TILs) of patients with melanomas. Genes encoding peptide antigens recognized by CTLs have been cloned from melanomas using these CTLs. However, either the presence of these CTLs or peptide antigens have been rarely reported in either adenocarcinoma of squamous cell carcinoma, two major human cancers needed for development of new treatment modalities. We have investigated HLA-A locus-restriction and tumor-specificity of IL-2 activated TILs from esohageal cancers, gastric cancers, colon cancers and lung cancers. The results showed the presence of HLA class lredtricted and tumor specific CTLs in TILs of esophageal cancers (Toh et al., Cell lmmunol. in press, 1997), gastric cancers (Hoshino et al., Int. J.Cancer in press, 1997), colon cancers (Gouhara et al., JJCR,in press, 1997) and non-small lung cancers (Seki et al., Cell lmmunolo. in press, 1997). These CTLs could be a tool for identification of genes encoding tumor-rejection antigens for development of cancer vaccines. Less