HCV多中和抗原表位嵌合病毒样颗粒免疫获得血清内中和抗体评价

Evaluation of neutralizing antibodies in serum immunized with virus-like particle chimerized HCV series of neutralizing epitopes

目的:纯化的HCV多中和抗原表位及HCV包膜蛋白E2嵌合的HBV S抗原病毒样颗粒免疫新西兰兔,测定免疫血清内的中和抗体。分析中和抗体对HCVpp感染Huh7.5的中和作用。方法:纯化的HCV多中和抗原表位及包膜蛋白E2嵌合的HBV S抗原病毒样颗粒(VLPs-MEp S、VLPs-E2S)10μg皮下接种新西兰兔,间隔2周共免疫3次,采集不同时间免疫兔血清,ELISA方法测定血清内的抗体,PBS组作为对照;制备1b型HCVpp,观察血清抗体对HCVpp感染Huh7.5的中和作用,对免疫血清保护性进行初步评价。结果:免疫后血清中产生一定水平的中和抗体,血清中和抗体测定VLPs-MEp S明显高于VLPs-E2S(P<0.05)。VLPs-MEp S与VLPs-E2S组均显著高于对照PBS组(P<0.01)。对HCVpp抑制作用,VLPs-MEp S高于VLPs-E2S组(P<0.05),最高中和率可达61.49%,二者均高于对照组(P<0.01)。结论:嵌合病毒样颗粒免疫新西兰兔后产生一定水平中和抗体,该中和抗体具有保护作用,为研发中和抗体表位疫苗奠定基础。

Objective：New Zealand rabbits were immunized withVLPs-MEpS,VLPs-E2S,and the levels of neutralizing antibodies in serum were determined.Methods： New Zealand rabbits were immunized with 10 g VLPs-MEpS and VLPs-E2S,serum was collected at diffferent time with a two-weeks interval.The neutralizing antibodies were determined by ELISA.HCV（type 1b） had been prepared and mixed with serum from immunized rabbit before infected Huh7.5 cell.The protection of neutralizing antibodies in serum was assessed.Results： Neutralizing antibodies had been induced in rabbit after immunized with VLPs-MEpS and VLPs-E2S.VLPs-MEpS group had higher titer of antibodies than that of VLPs-E2S group（P〈0.05）,both group had higher titer of antibodies than that of control groups significantly（P 〈0.01）.VLPs-MEpS group had higher neutralization than that of VLPs-E2S group（P 〈0.05）,the highest neutralization rate was 61.49%.Both groups were higher than control group notably（P 〈0.01）. Conclusion：Protective neutralizing antibodies have been induced in New Zealand rabbit after immunized with VLPs-MEpS and VLPs-E2S.It’s the basement for development of neutralizing antibodies vaccine.