HCV中和抗原表位与HBV S抗原嵌合病毒样颗粒诱导的中和抗体应答及保护评价

Evaluation for protection of the neutralization antibody induced by particle of HBV S antigen chimerized with HCV neutralizing epitopes in mice

目的用纯化的嵌合HCV中和抗原表位的HBV VLPs免疫Balb/c小鼠,测定小鼠血清中的中和抗体,并观察免疫血清对HCVpp和HCVcc感染Huh7.5的抑制作用。方法 500 ng纯化的嵌合病毒样颗粒抗原,免疫Balb/c小鼠3次,每2周尾静脉采血,用合成的表位肽和HBs Ag阳性血清作为包被抗原检测小鼠血清中抗体。免疫血清和HCVpp及HCVcc预处理,观察其对HCVpp和HCVcc感染Huh7.5细胞的抑制作用。结果 Balb/c小鼠体内诱导出HBs Ab和一定水平的针对HCV中和抗原表位的中和抗体,混合VLPs免疫诱导的中和抗体水平高于单一VLPs诱导产生的中和抗体。用免疫小鼠血清预处理的HCVpp和HCVcc感染Huh7.5能力明显低于对照组。结论嵌合HCV中和抗原表位的HBV VLPs免疫小鼠可诱导产生针对中和抗原表位的中和抗体,该中和抗体能抑制HCVpp和HCVcc对Huh7.5的感染,具有一定的保护作用。

This study designed to evaluate the infection inhibitory effects of neutralization antibody from Balb/c immunized by HBV VLPs chimerized with HCV neutralization epitopes, by analyzing its effects on the infection of HCVpp and HCVcc to Huh7.5 cells. Balb/c mice was immunized by 500 ng purified chimerized HCV HBV S VLPs,and serum was collected from caudal vein every twice weeks. The neutralization antibody in serum was determined by the synthesis epitopes peptide and HBs Ag. Huh7.5 cell was infected by mouse serum-pretreated HCVpp and HCVcc and infection inhibition was analyzed by FCM and FFU. Neutralization antibody for HCV epitopes and HBs Ab were induced in mice immunized by different VLPs. Data showed that higher antibody levels were induced in mice immunized with mixture VLPs than that in mice immunized with single VLPs; the ability of infection to Huh7.5 for HCVpp and HCVcc pretreated with mouse serum was significant lower than that of control groups. In conclusion, neutralization antibody is induced in mice immunized by VLPs chimerized with HBV S and HCV neutralizing epitopes, which can protect Huh7.5 cells from infection of HCVpp and HCVcc.