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Identification of a cluster of tumor antigens recognized by CTh of colon cancer patients

结肠癌患者 CTh 识别的一组肿瘤抗原的鉴定

基本信息

批准号：
14570526
负责人：
SHICHIJO Shigeki
金额：
$ 2.18万
依托单位：
Kurume University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Background : Colon cancer is one of major malignant tumors to which development of new treatment modality is needed. To provide scientific basis of specific immunotherapy of colon cancer, this study focused on identification of a cluster of tumor antigens and their epitopes recognized by HLA-A2-restricted and tumor-reactive cytotoxic T lymphocytes (CTLs) generated from tumor-infiltrating lymphocytes (TIL) of a colon cancer patient.Methods : The HLA-A2-restricted and tumor-reactive OK-Cm line was established from TILs of a patient with colon cancer. A gene-expression cloning method was used to identify genes coding for tumor antigens recognized by the OK-CTLd subline. To identify CTL-directed epitopes, the CTLs were incubated for 18 hr with T2 cells pre-pulsed with each peptide at different doses for 2 hr followed by harvesting of supernatant to measure IFN-γ by ELISA.Results : We identified 27 genes, including 9 unique genes. Thirteen (56.5%) of the 23 genes with known or presumed functions encoded proliferation-related proteins. The others mainly consisted of genes encoding transporters. Among 393 peptides with HLA-A2 binding motifs encoded by the 27 genes, 68 peptides were recognized by the parental CTLs, and 26 peptides among them were also recognized by CTLs in the circulation of colon cancer patients. Further, 16 of these 26 peptides possessed the ability to induce HLA-A2 restricted and peptide-specific CTLs cytotoxic to colon tumor cells in peripheral blood mononuclear cells of colon cancer patients. In the other hands, 5 and 7 peptides encoded by 2 and 4 genes, respectively, were identified with recognized by HLA-B46 and -A31 restricted CTL.Conclusion : These antigens and peptides could be appropriate in use for specific immunotherapy of HLA-A2^+ colon cancer patients.

背景：结肠癌是重要的恶性肿瘤之一，需要发展新的治疗方式。为了给结肠癌的特异性免疫治疗提供科学依据，本研究对结肠癌患者肿瘤浸润淋巴细胞（TIL）产生的hla - a2限制性和肿瘤反应性细胞毒性T淋巴细胞（ctl）识别的肿瘤抗原及其表位进行了鉴定。方法：从1例结肠癌患者的TILs中建立hla - a2限制性、肿瘤反应性的OK-Cm细胞系。采用基因表达克隆的方法鉴定了OK-CTLd亚系识别的肿瘤抗原编码基因。为了鉴定ctl导向的表位，将ctl与T2细胞孵育18小时，每个肽以不同剂量预脉冲2小时，然后收集上清，通过ELISA检测IFN-γ。结果：共鉴定出27个基因，其中独特基因9个。已知或推测功能的23个基因中有13个（56.5%）编码增殖相关蛋白。其他主要由编码转运蛋白的基因组成。在27个基因编码的393个具有HLA-A2结合基序的肽段中，68个肽段被亲本ctl识别，其中26个肽段也被结肠癌患者循环中的ctl识别。此外，这26个多肽中的16个具有诱导HLA-A2限制性和多肽特异性ctl对结肠癌患者外周血单个核细胞中的结肠肿瘤细胞具有细胞毒性的能力。另一方面，鉴定出由2个和4个基因编码的5个和7个肽段分别被HLA-B46和-A31限制性CTL识别。结论：这些抗原和多肽可用于HLA-A2^+结肠癌患者的特异性免疫治疗。