溶酶体靶向的汉坦病毒包膜糖蛋白DNA疫苗的体液免疫评价和攻毒保护效果

摘要

目的:构建嵌合溶酶体相关膜蛋白(LAMP)的汉坦病毒糖基化蛋白DNA疫苗并对其免疫进行评价.方法:利用前期实验构建的重组质粒pVAX-Gn、pVAX-Gc、pVAX-LAMP/Gn和pVAX-LAMP/Gc以及inactivated疫苗免疫BALB/c小鼠,分别通过间接 ELISA 和中和试验检测免疫小鼠血清中的特异性抗体和中和抗体;攻毒实验检测小鼠体内的保护效力.结果:间接ELISA结果显示,pVAX-LAMP/Gc组特异性抗体滴度最高,依次为pVAX-Gc、pVAX-LAMP/Gn、pVAX-Gn与inactivated疫苗组;中和结果显示,LAMP组均优于相应传统DNA疫苗组,且抗体效价均优于Inactived vaccine组;攻毒实验显示,DNA疫苗和inactivated疫苗组小鼠体内无汉坦病毒特异性抗原检出.结论:LAMP分子的嵌合DNA疫苗可诱导更高水平的抗体,在小鼠体内有较好的保护效力,这一靶向策略有望成为提高DNA疫苗效价的有效方式.%Objective:To build the DNA vaccine encoding hantavirus glycoprotein fused with lysosome-associated membrane protein (LAMP) and assess its immune response.Methods:BALB/c mice were immuned with the previous experimental expressing purified recombinant plasmids pVAX-Gn,pVAX-Gc,pVAX-LAMP/Gn,pVAX-LAMP/Gc,and inactivated vaccine.Indirect ELISA and neutralization test was used to detect specific antibody and neutralizing antibody in the serum of mice.The mice were tested to detect the protective effects in vivo.Results:Indirect ELISA results showed that the pVAX-LAMP/Gc group was the highest,followed by pVAX-Gc,pVAX-LAMP/Gn,and PVAX-Gn,and inactivated vaccine group.In neutralization test,there were significantly higher serum antibodies in LAMP group than those in conventional DNA group,which were higher than the inactivated vaccine group.The mice immuned had good protective effect in vivo without the specific antigen of hantavirus in vivo.Conclusion:Chimeric DNA vaccines induced higher levels of antibody in BALB/c mice and produced better protective efficacy,which illustrate the targeting strategy is expected to be an effective way to improve the DNA vaccine efficacy.