Human hepatoma antigen analyzed by genetic engineering method

利用哺乳动物细胞系表达载体pcDNA3，转染了HLA-A24基因缺失的人肝癌细胞株Hep.3B2.1-7。1以便在其细胞表面携带内源性抗原肽。克隆的表达Hep.3B2.1-7的HLA-A24酸性洗脱液（Hep.3B.A24）经FPLO分离后，与野生型Hep.3B2.1-7和人工转染的Hep.3B2.1-7不同，有7个显著峰。7个片段中有1个片段在第2位(L)和第10位(K)具有HIA-A24的锚定基序，其余6个片段中有2个组蛋白H2A、1个巨噬细胞迁移抑制因子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。总之，本研究有可能阐明一种新的HLA- a24结合肽和HLA。通过基因工程方法实现分子的表达，将为肿瘤抗原肽的研究开辟一条新的途径。

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.