摘要
目的通过脂质体介导乙肝病毒核心抗原(HBcAg)基因转染人体内专职抗原提呈的树突状细胞(DC)的方法,使HBcAg基因在树突状细胞中表达,构建树突状细胞疫苗。方法分离健康人外周血单个核细胞,在粒-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素-4(IL-4)的诱导下培养DC。在培养的第5天,应用脂质体PEI介导将含有编码HBcAg 基因的质粒(pJW4303／HBc)转染入DC,再以免疫印迹技术(Western-Blotting)验证其表达产物。结果经以质粒pJW4303 为真核表达载体的HBcAg目的基因转染后的树突状细胞有效表达了HBcAg抗原。结论经该方法转染后的DC目的基因可有效表达HBcAg,为以DC为基础的免疫治疗奠定了新的基础。
Objective To increase the immune response which anti-HBVthrough the method that the plasmid vector contained HBcAg gene transfer the dendritic cells（DCs） which is the professional antigen processing cell in the human being.Methods Peripheral blood mononuclear celIs（PBMC） were isolated from whole blood by settling process on HES,the adherent cells were cultured in the medium RPMI-1640 contained GM-CSF and IL-4.On day 5, lipofectin-mediated gene transfection method was used to transfected DCs.The expression of HBcAg gene was confirmed by Western-Blotting.Results The expression of HBcAg from DCs which was transferred was found by Western-Blotting.Conclusion The method maked DCs which were transferred expressed HBcAg effectly ,it provide a new kind of way for the immune treatment based on DCs.
出处
《中国血液流变学杂志》
CAS
2006年第1期23-25,共3页
Chinese Journal of Hemorheology
基金
江苏省"135重点学科"基金江苏省"333工程"基金资助
