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蛋白的自身抗体：我们用酶联免疫吸附试验检测了188例非小细胞肺癌患者的抗p53自身抗体。p53抗体阳性率为18%（34/188）。鳞状细胞癌患者或III-IV期疾病患者的p53抗体发生率分别显著较高。有p53蛋白异常表达的肿瘤患者p53抗体阳性率高于无p53蛋白异常表达的肿瘤患者（28%vs14%）。然而，我们的数据没有表明p53抗体作为NSCLC复发或预后的标志物的临床有用性。2）HLA-A2阳性非小细胞肺癌患者中法师-3基因表达的频率：表达法师-3的肿瘤细胞被细胞毒性T淋巴细胞以HLA A1和A2限制的方式消除。因此，其肿瘤共表达HLA-A1或-A2和法师-3的癌症患者可以是疫苗治疗的潜在候选者。 关于我们 或者，这种肿瘤可能在成为临床上明显的肿瘤之前已经在体内消除。我们评估了57例肺癌中HLA-A1或-A2表达与法师-1或-3基因表达之间的关系。法师3阳性24例（42%）。HLA-A2蛋白表达阳性17例（30%）。HLA-A2患者中法师-3表达的频率为5/17（29%），略低于无HLA-A2表达的患者18/38（47%）（p = 0.17）。我们的结论是，没有明确的证据消除肺癌共表达HLA-A2和法师-3在体内。约10%（5/55）的日本肺癌患者是基于法师-3的免疫治疗的潜在候选者。3）使用树突状细胞（DC）诱导HLA A24限制性p53特异性CTL：使用6种p53衍生肽作为抗原，并且使用来自健康志愿者PBMC的DC作为APC，我们尝试诱导p53特异性CTL。然而，6个肽均未成功诱导CTL。虽然我们用未成熟DC代替成熟DC或加入β2微球蛋白等方法进行了改进，但均无效。尽管存在肽本身的选择不合适的可能性，但存在CTL的前体频率低于检测水平的另一种可能性，因为我们仅使用来自健康供体的PBMC。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