Simultaneous cellular and humoral immune response against mutated p53 in a patient with lung cancer
Simultaneous cellular and humoral immune response against mutated p53 in a patient with lung cancer
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DOI：
10.4049/jimmunol.172.8.4844
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发表时间：
2004-04-15
影响因子：
4.4
通讯作者：
Yasumoto, K
中科院分区：
文献类型：
--
作者：
Ichiki, Y;Takenoyama, M;Yasumoto, K
我们最近使用 SCID 小鼠和异种移植非小细胞肺癌系统鉴定了几种可被肿瘤浸润 B 淋巴细胞衍生抗体识别的 Ag。其中一个已鉴定的 Ag 发生 p53 突变，产生点突变，导致 158 号密码子从 Arg 变为 Leu。本研究的目的是确定针对突变 p53 的细胞免疫是否与不道德免疫同时存在于同一患者中。根据来自患者的已建立癌细胞系A904L的HLA I类结合基序，合成了两种不同的Nona肽（突变的p53（150）和p53（155）肽），包括源自p53的突变氨基酸。每周用肽刺激患者的纵隔淋巴结淋巴细胞。突变的p53(155)肽刺激的淋巴细胞对用突变的p53(155)肽和A904L脉冲的自体EBV转化的B细胞显示出特异性细胞毒性。建立了 HLA-Cw*0702 限制性突变的 p53(155) 肽特异性 CTL 克隆，并分析了其 TCR 使用情况。使用 CDR3 特异性引物进行克隆型 PCR 应用于含有肿瘤浸润淋巴细胞的肿瘤组织。在肿瘤组织中发现PCR特异性扩增。这些结果表明，肿瘤组织中不仅存在产生针对p53蛋白的特异性Ab的B淋巴细胞，而且还存在针对自体肺癌细胞中表达的突变p53的CTL。这种方法可以让我们更好地了解癌症患者中 T 细胞和 B 细胞针对相同肿瘤 Ag 的免疫机制。
We recently identified several Ags recognized by tumor-infiltrating B lymphocyte-derived Ab using SCID mice and a xenografted non-small cell lung cancer system. One of these identified Ags was mutated p53 with a point mutation resulting in the alteration of codon 158 from Arg to Leu. The aim of this study was to ascertain whether cellular immunity against mutated p53 exists in the same patient together with Immoral immunity. Two different nona peptides (mutated p53(150) and p53(155) peptides), including a mutated amino acid derived from p53, were synthesized according to the binding motif of HLA class I of the established cancer cell line A904L from the patient. Mediastinal lymph node lymphocytes of the patient were stimulated weekly with the peptides. The mutated p53(155) peptide-stimulated lymphocytes showed specific cytotoxicity against both autologous EBV-transformed B cells pulsed with mutated p53(155) peptide and A904L. The mutated p53(155) peptide-specific CTL clone in an HLA-Cw*0702 restriction was established and analyzed for its TCR usage. Clonotypic PCR using CDR3-specific primers was applied to the tumor tissue containing the tumor-infiltrating lymphocytes. The specific amplification of PCR was found in the tumor tissue. These results demonstrated that not only B lymphocytes producing specific Ab against the p53 protein, but also CTL against mutated p53, expressed in autologous lung cancer cells exist in the tumor tissue. This approach may allow us to better understand the mechanisms of T and B cell immunity against the same tumor Ag in cancer patients.