Human hepatoma antigen analyzed by genetic engineering method

基因工程方法分析人肝癌抗原

• 批准号: 09670181
• 负责人: TAKAMI Tsuyoshi
• 金额: $ 1.92万
• 依托单位: Gifu 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: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670181/
• 关键词: Hepatoma; HLA-A24; genetic engineering; Hpe.3B2.1-7; Acid elution; FPLC; Peptide; 肝癌細胞; 腫瘍抗原; Hep-3B細胞; 抗HLA-A24抗体; ペプチド・シークエンス; 肝癌腫瘍抗原; Hep3B; cDNA; 単クローン抗体

Human hepatoma cell line, Hep.3B2.1-7 that has no HLA-A24 gene expression, has been transduced of HLA-A24 cDNA by using mammalian cell line expression vector pcDNA3 .1 in order to carry the endogenous antigenic peptides on its cell surface. The acidic eluate of cloned HLA-A24 expressing Hep.3B2.1-7 (Hep.3B.A24) demonstrated seven significant peakes that were different from wild type Hep.3B2.1-7 and moke-transfected Hep.3B2.1-7 after FPLO fractionations.One out of seven peptide fraction had anchoring motief for HIA-A24 at second (L) and tenth (K) positions, while two histon H2A, one HLA-A68 binding peptide of macrophage migration inhibiting factor, one HIA-B35 binding peptide, and two unknown peptides were clarified from remaining six fractions.The homology research of HLA-A24 binding peptide obtained in present study demonstrated no homologous protein in data base.In conclusion, present study is possible to clarify a novel HLA-A24 binding peptide, and HLA.molecules enforced to express by genetic engineering method will open a new avenue for the research in tumor-antigenic peptides.

不具有HLA-A24基因表达的人肝癌细胞系Hep.3B2.1-7已通过使用哺乳动物细胞系表达载体pcDNA3.1转导HLA-A24 cDNA，以便在其细胞表面携带内源抗原肽。表达 Hep.3B2.1-7 (Hep.3B.A24) 的克隆 HLA-A24 的酸性洗脱液在 FPLO 分级后显示出与野生型 Hep.3B2.1-7 和 moke 转染的 Hep.3B2.1-7 不同的 7 个显着峰。 七个肽级分中的一个在第二个 (L) 和第十个 (K) 处具有 HIA-A24 的锚定基序 其余6个组分中澄清了2个组蛋白H2A、1个巨噬细胞迁移抑制因子的HLA-A68结合肽、1个HIA-B35结合肽和2个未知肽。本研究获得的HLA-A24结合肽的同源性研究表明数据库中没有同源蛋白。综上所述，本研究有可能澄清一种新的蛋白。 HLA-A24结合肽以及通过基因工程方法强制表达的HLA分子将为肿瘤抗原肽的研究开辟新的途径。