脂质转染剂增强基因疫苗诱导的免疫应答效力

LipofectAMINE coated hepatitis C virus core gene vaccine promotes the efficacy of immune responses

目的 研究脂质转染剂 (L ipofect AMINE)提高丙型肝炎病毒 (HCV)核心 (C)基因疫苗的抗病毒免疫应答效力 .方法 人工构建包含 HCV C基因片段的真核表达载体 pc D-NAHCV- C,在证实其可以在真核细胞中表达之后 ,将其用脂质转染剂包裹 ,形成 L ipofect AMINE- pc DNAHCV- C脂质混合物 ,将该混合物或单纯 pc DNAHCV- C直接注射 BAL B/ c小鼠股四头肌 ,以空载体 pc DNA3做对照 ,EL ISA法检测血清中抗体产生水平 .结果 12只实验组小鼠 ,初次免疫 2 wk后 ,血清中均出现了 HCV C抗体 ,而 L ipofect AMINE- pc D-NAHCV- C混合物免疫组 ,抗体表达值明显高于单纯 pc D-NAHCV- C免疫组 (P<0 .0 5 ) .结论 脂质转染剂可以促进基因疫苗的摄取并增强其诱导的抗病毒免疫应答的效力 .

AIM To promote the efficacy of LipofectAMINE coated recombinant plasmid containing hepatitis C virus (HCV) core gene inductive immune responses. METHODS The HCV core gene coding region was inserted into the eukaryotic expressive plasmid pcDNA3, then the recombinant plasmid pcDNAHCV-C was constructed and expressed transiently with LipofectAMINE in the SP2/0 cells. After purification, these plasmid directly or encapsuled with Lipofect-AMINE were injected into BALB/c mice. HCV core antibody from immunized mice could be detected by ELISA. RESULTS The enzyme-cutting identification showed that HCV core gene fragment had been cloned into pcDNA3 eukaryote vectors. The expressed products could combine with patient serum of HCV C antibody positive, and HCV C antibody was time dependence, and HCV C antibody expressing value from encapsuled with LipofectAMINE HCV core gene vaccine-immunized mice were higher than from control mice ( P < 0.05 ). CONCLUSION Liposome technique in combination with gene vaccine can promote the efficacy of immune responses in BALB/c mice.