Development of immunotherapy for lung cancer utilizing p53 specific cytotoxic T lymphocytes

1)非小细胞肺癌患者血清中抗P53蛋白自身抗体：采用酶联免疫吸附试验检测188例非小细胞肺癌患者血清中抗P53蛋白自身抗体。188例患者中有34例(18%)P53抗体阳性。鳞状细胞癌或III-IV期疾病患者的P53抗体发生率分别显著高于其他患者。有P53蛋白异常积聚的肿瘤患者P53抗体阳性率高于无P53蛋白异常的肿瘤患者(28%vs14%)。2)非小细胞肺癌患者中MAGE-3基因表达的频率：表达MAGE-3的肿瘤细胞被表达MAGE-3的细胞毒性T淋巴细胞以HLAA1和A2限制的方式清除。因此，肿瘤共表达HLA-A1或-A2和MAGE-3的癌症患者可能成为疫苗治疗的潜在候选者。…更有可能的是，这种肿瘤在成为临床显性肿瘤之前可能已经在体内消除了。我们研究了57例肺癌组织中人类白细胞抗原A1和A2的表达与MAGE-1和MAGE-3基因表达的关系。MAGE-3阳性24例(42%)。有17例(30%)在蛋白水平表达了HL A-A2。HLA-A2组MAGE-3表达频率为5/17(29%)，略低于未表达组的18/38(47%)(P=0.17)。我们的结论是，没有明确的证据表明在体内消除了共表达人类白细胞抗原A2和MAGE-3的肺癌。大约10%(5/55)的日本肺癌患者是基于MAGE-3的免疫治疗的潜在候选者。3)用树突状细胞(DC)诱导HLAA24限制性P53特异性CTL：以6个P53来源的多肽为抗原，以健康志愿者PBMC来源的DC为APC，我们尝试诱导P53特异性CTL。然而，这6个多肽均不能成功诱导CTL。虽然我们用未成熟的DC代替成熟的DC或加入β-2微球蛋白对方法进行了改进，但均未成功。虽然有一种可能是多肽本身的选择不合适，但另一种可能性是CTL的前体频率低于检测水平，因为我们只使用健康供者的PBMC。较少

1) Auto-antibodies against p53 protein in sera of NSCLC patients: We examined 188 consecutive patients with NSCLS for auto antibodies against p53 by enzyme-linked imunosorbent assay. p53 antibodies were detected in 34 patients of 188 (18%). Patients with squamous cell carcinoma or those with stage III-IV disease had a significantly higher incidence of p53 antibodies, respectively. Prevalence of p53 antibodies in patients with tumors having abnormal p53 protein accumulation was higher than that in patients without p53 abnormality (28% vs 14%). However, our data did not indicate clinical usefulness of p53 antibodies as a marker for relapse or prognosis of NSCLC.2) Frequency of MAGE-3 gene expression in HLA-A2 positive patients with non-small cell lung cancer: Tumor cells expressing MAGE-3 are eliminated by cytotoxic T lymphocytes in HLA A1 and A2 restricted fashion. Therefore cancer patients whose tumor co-expresses HLA-A1 or -A2 and MAGE-3 can be potential candidates of vaccine therapy. … More Alternately, such tumors might have eliminated in vivo before becoming clinically overt tumor. We assessed the relationship between HLA-A1 or -A2 expression and MAGE-1 or -3 gene expression in 57 lung cancers. MAGE-3 were detected in 24 (42%). HLA-A2 was expressed in 17 (30%) at the protein level. The frequency of MAGE-3 expression in HLA-A2 patients was 5/17 (29%), somewhat lower than that of patients without HLA-A2 expression, 18/38 (47%) (p = 0.17). We conclude that there is no clear evidence for elimination of lung cancers co-expressing HLA-A2 and MAGE-3 in vivo. Approximately 10 percent (5/55) of Japanese lung cancer patients are potential candidates for MAGE-3 based immunotherapy.3) Induction of HLA A24 restricted p53 specific CTL using dendritic cells(DC): Using 6 p53-derived peptides as antigens and DC derived from PBMC of healthy volunteers as APC, we tried to induce p53 specific CTL. However, none of the 6 peptide successfully induced CTL. Although we modified the method by using immature DC instead of mature DC, or by adding β2 microglobulin but in vain. Although there is a possibility that the selection of peptides itself was not appropriate, another possibility exists that precursor frequency of CTL was below the detection level since we solely used PBMC from healthy donors. Less