Development of viral antigen specific immunotherapy for Epstein-Barr virus related gastric cancer.

开发针对 Epstein-Barr 病毒相关胃癌的病毒抗原特异性免疫疗法。

• 批准号: 12671249
• 负责人: UEDA Yuji
• 金额: $ 1.15万
• 依托单位: Kyoto Prefectural University 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671249/
• 关键词: EBV (Epstein-Barr Virus); Gastric Cancer; Tumor Antigen Peptide; Tumor Specific Immunotherapy; Cancer vaccine Therapy; LMP2(Latent Membrane Protein 2)

(1) Cytotoxic T cell(CTL) induction with latent membrane protein 2(LMP2) epitope peptide -loaded dendritic cells(DCs).We synthesized the LMP2 peptide, TYGPVFMCL(HLA-A24 restricted), and established CTL line by stimulating PBMCs from HLA-A24 positive healthy donors in vitro with LMP2 peptide-loaded DCs. This CTL could lyse the target cells pulsed with this peptide, but couldn't lyse LMP2-CDNA transformed target cells.(2) CTL induction with autologous EBV transformed B cell line(LCL).In order to establish LCL, concentrated supernatant from B95-8 cultures, a human type1 EBV-transformed marmoset B-cell line, was added to a healthy donor's PBMCs. PBMCs from healthy donors were stimulated in vitro with autologous LCL. This LCL-induced CTL recognized the full repertoire of EBV latency antigens. We selected LMP2-specific CTL line after 5lCr release assay.(3) Detection of EBV-DNA by polymers chain reaction (PCR).In order to detect EBV related gastric cancers from patients underwent gastorectomy in our institute, we performed PCR and southern hybridization. After that, we tried to establish LMP2 positive cell lines of gastric cancer which could be useful targets for CTL-based immunotherapy.

(1)为探讨LMP 2抗原表位肽负载树突状细胞（DCs）诱导细胞毒性T细胞（CTL）的作用，我们人工合成了LMP 2抗原表位肽TYGPVFMCL（HLA-A24限制性），并将其负载DCs，体外刺激HLA-A24阳性健康人PBMCs，建立了CTL细胞系。该CTL能裂解经该肽脉冲的靶细胞，但不能裂解经LMP 2-cDNA转化的靶细胞。(2)用自体EBV转化的B细胞系（LCL）诱导CTL为了建立LCL，将来自B 95 -8培养物（人1型EBV转化的绒猴B细胞系）的浓缩上清液加入健康供体的PBMC中。来自健康供体的PBMC在体外用自体LCL刺激。这种LCL诱导的CTL识别EBV潜伏抗原的全部库。经51 Cr释放试验筛选出LMP 2特异性CTL细胞株。(3)聚合物链反应（PCR）检测EBV-DNA为检测我所胃切除术后患者中与EBV相关的胃癌，我们采用PCR和Southern杂交技术。在此基础上，我们尝试建立LMP 2阳性胃癌细胞系，为CTL免疫治疗提供有用的靶点。

项目成果

上田祐二: "進行癌治療とワクチン療法"癌治療と宿主. 12(2). 179-186 (2000)

Yuji Ueda：“先进的癌症治疗和疫苗治疗”癌症治疗和主持人179-186（2000）。

Tsuyosi Itoh, Yuji Ueda, et al.: "Immunotherapy of Solid Cancer using Dendritic Cells Pulsed with the HLA-A24-restricted Peptide of Carcinoembrionic Antigen"Cancer Immunol Immunother. 50(in press). (2002)

Tsuyosi Itoh、Yuji Ueda 等人：“使用用癌胚抗原的 HLA-A24 限制性肽脉冲的树突状细胞进行实体癌的免疫治疗”癌症免疫免疫疗法。

上田祐二: "進行消化器癌に対する癌ワクチン療法のtumor dormancy therapyとしての可能性"日本消化器外科学会雑誌. 34(4). (2001)

Yuji Ueda：“癌症疫苗疗法作为晚期胃肠道癌症的肿瘤休眠疗法的可能性”，日本胃肠外科学会杂志 34（4）（2001）。

Tsuypshi Itoh, Yuii Ueda. Hisakazu Yamagishi. et al.: "lmmunotherapy of Solid Cancer using Dendritic Cells Pulsed with the HLA-A24-restricted Peptide of Carcinoembrionic Antigen"Cancer Immunol Immunother. 50(accepted (in press)). (2002)

伊藤刚、上田由衣。

山岸久一: "癌のワクチン療法(癌の治療戦略-現況と展望-)"外科治療. 82. 691-695 (2000)

Hisaichi Yamagishi：“癌症疫苗治疗（癌症治疗策略 - 现状和前景）” Surgical Treatment。 82. 691-695 (2000)。