糖基化磷脂酰肌醇锚定型人B7-1基因重组蛋白锚定自体肿瘤细胞膜提高抗肿瘤免疫反应的实验研究

目的研制抗肿瘤免疫治疗的新疫苗。方法用含目的基因的质粒pcDNA3-GPI-B7-1转染CHO/DHFR+细胞,提取膜蛋白GPI-B7-1,经Western blot鉴定后,用蛋白转化法锚定在肿瘤细胞膜上,免疫C57BL/6小鼠后,取小鼠脾细胞进行T细胞扩增和CTL功能检测,并检测细胞培养液和小鼠血清中IL-2、TNF-α和IFN-γ水平。结果用GPI-B7-1修饰的肿瘤细胞膜免疫小鼠后可诱发肿瘤特异性T细胞扩增和CTL活性增强,细胞培养液中细胞因子检测为阳性。结论该疫苗能激发小鼠的免疫功能,具有一定的保护小鼠免受肿瘤细胞侵袭的作用。

Experimental Study on Enhancement of Immune Response against Tumor by Anchoring Glycosyl-phosphatidylinositol-B7-1 onto Tumor Cell Membrane

Objective To study the role of anchoring glycosyl-phosphatidylinositol(GPI)-B7-1 onto tumor cell membrane in immune response against tumor and provide a basis for developing a novel vaccine for immunotherapy against tumors.Methods Transfect CHO/DHFR~+ cells with recombinant plasmid pcDNA3-GPI-B7-1,extract membrane protein GPI-B7-1 and identify by Western blot.Anchor GPI-B7-1 onto tumor cell membrane and immunize to C57BL/6 mice.Isolate the spleens of mice for proliferation of T cells and analysis of CTL function.Determine the IL-2,TNF-α and IFN-γ levels in splenocyte culture and murine serum.Results Tumor-specific T cell proliferation and CTL were induced in the mice immunized with GPI-B7-1-anchored tumor cell membrane.The detection result of cytokines in splenocyte culture was positive.Conclusion(The GPI-B7-1-anchored) tumor enhanced the immune response of mice and showed a certain protection against challenge with tumor cells.