超抗原SEA的GPI锚定修饰和在CHO-dhfr^-细胞膜上的表达

Construction of GPI-anchored Superantigen SEA and Expression in CHO-dhfr-Cells Membrane

将GPI锚定修饰的免疫分子直接转移到肿瘤细胞膜上 ,是研究治疗性肿瘤疫苗的一种有潜力的新策略。通过将从衰变加速因子 (DAF)来源的GPI修饰性信号序列与SEA嵌合 ,获得了GPI修饰的SEA分子 ,构建的真核表达载体pCI GPI-SEA ,用脂质体方法转染到CHO dhfr-细胞中 ,并用氨甲喋呤 (MTX)进行筛选 ,细胞免疫荧光分析证实 ,SEA能够在细胞膜上表达。上述GPI锚定修饰的SEA ,可用于进一步研究治疗性肿瘤疫苗。

Direct modification of tumor cell membranes by transfer of a glycosyl-phosphatidylinositol (GPI)-anchored immunological molecules has been proposed as a novel potentially useful strategy to development of therapeutic tumor vaccines.In this study,Glycosylphosphatidylinositol (GPI)- anchored variants of superantigen SEA were produced via chimerization with alternative GPI-modification signal sequences from decay-accelerating factor (DAF).Eukaryotic expression vector pCI/GPI-SEA was transfected into CHO-dhfr- cell with lipofectine-mediated gene transfer technique,and the transfectants were selected with methotrexate (MTX).Immunofluorescence staining assay .suggests that SEA expression in CHO cell membranes.GPI-anchored SEA may be prepared for use as therapeutic tumor vaccines.;