Development of cancer vaccine with MAGE gene products.

MAGE-1、MAGE-2、MAGE-3、MAGE-4、MAGE-6和MAGE-12基因在多种肿瘤中均有表达，但在正常细胞或正常组织中不表达。MAGE-1和MAGE-3多肽目前被用作癌症患者的疫苗，12名转移性黑色素瘤患者中有3名对MAGE-3多肽有反应。因此，MAGE蛋白有可能成为治疗人类白细胞抗原A1癌患者的潜在疫苗。然而，在应答患者的外周血中未检测到CTL，也没有可用的监测方法来了解MAGE疫苗的效果。此外，MAGE家族蛋白的生物学作用仍有待研究。我们研究了多种癌症患者血清中MAGE蛋白家族之一的MAGE-4蛋白的水平。在不同组织学类型的癌症患者(肺癌、头颈癌和肝细胞…)的MAGE-4肿瘤细胞株上清液和血清中均检测到非降解形式的MAGE-4蛋白更多的癌症。例如，肺癌患者血清MAGE-4蛋白水平(n=100，平均为1.17 ng/ml)显著高于良性肺部疾病患者(n=80，平均为0.33 ng/ml)和先天性心脏病患者(n=68，平均为0.32 ng/ml)(Shichijo等，JJCR，1977年出版)。100例癌症患者中有34例血清MAGE-4水平高于临界值(1.15 ng/ml)，其他组无一例达到临界值。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，手术切除肿瘤导致血清MAGE-4蛋白水平下降，复发与其再次升高有关(Iwamoto等人，Int。《癌症杂志》，出版，1997)。此外，在肝细胞癌和丙型肝炎病毒阳性的肝硬变患者中，血清MAGE-4水平也显著升高，后者是肝细胞癌的高危人群。这些信息可能有助于更好地了解MAGE蛋白的生物学作用，血清MAGE-4蛋白水平的测定可能有助于检测不同组织类型的MAGE-4癌症。在黑色素瘤患者的自体肿瘤细胞或IL-2激活的肿瘤浸润性淋巴细胞(TIL)刺激的外周血单个核细胞(PBMC)中，观察到HLAI类限制性和肿瘤特异性CTL。利用这些CTL已从黑色素瘤中克隆出CTL识别的多肽抗原编码基因。然而，无论是这些CTL的存在还是多肽抗原的存在，在鳞状细胞癌这两种人类主要癌症中都很少有报道，这是开发新的治疗方式所必需的。我们研究了食道癌、胃癌、结肠癌和肺癌患者IL-2激活的TIL的HLA-A位点限制性和肿瘤特异性。结果表明，食道癌患者TIL中存在人类白细胞抗原(HL A)类特异性和肿瘤特异性CTL(Toh等人，细胞免疫)。出版，1997)，胃癌(Hoshino等人，Int.结肠癌(Gouhara等人，JJCR，出版社，1997)和非小肺癌(Seki等人，细胞免疫)。在媒体上，1997)。这些CTL可以作为一种工具来识别编码肿瘤排斥抗原的基因，用于开发癌症疫苗。较少

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