丙型肝炎病毒重组蛋白的免疫保护性

目的 研究两种HCV重组蛋白联合免疫小鼠所诱导的免疫应答及免疫保护作用.方法 用两种重组蛋白HCV-T和HCV-第一高变区多片段重组融合蛋白(F4HVR1)分别与联合免疫BALB/c小鼠,共免疫3次,用ELISA方法测定血清特异性抗体;末次免疫后14 d处死5只小鼠,分离小鼠脾细胞.体外检测IFN-γ、IL-4和行CTL杀伤实验;剩余的小鼠背部皮下注射1.0×106个SP2/O-NS3细胞,观察其保护作用.组间均数差异采用LSD-t检验.结果 与PBS组相比,用HCV-T和HCV-F4HVR1联合免疫诱导了针对HCV-F4HVR1的特异性的IgG(t=3.815,3.762,P＜0.05)、高水平的HCV-NS3特异性的CTL效应(t=3.971,P＜0.05)和高水平的IL-4(t=3.813,3.426,3.671,P＜0.05)和IFN-γ(t=3.512,3.417,P＜0.05)的分泌.结论 用HCV-T和HCV-F4HVR1联合免疫小鼠可诱导出高水平的特异性体液免疫和细胞免疫,能有效地预防SP2/O-NS3细胞的攻击.

Immunoprotection analysis of hepatitis C virus recombinant protein

Objective To investigate immune responses and protective effect induced by two recombinant proteins of hepatitis C virus(HCV)in BALB/c mice.Methods BALB/c mice were immunized with recombinant proteins HCV-T and(or)HCV-F4HVR1 three times.Specific antibodies in sera were tested by enzyme-linked immunosorbent assay(ELISA).Five mice were sacrificed after 14 days of the last immunization.Splenic cells were isolated and levels of interferon(IFN)-γ,interleukin(IL)-4 and cytotoxic T lymphocyte(CTL)cytotoxicity assay were measured in vitro.The remaining mice were subcutaneously injected with 1.0×106 SP2/0-NS3 cells on the back to investigate the protective effects.The differences of means between groups were compared by LSD-t test.Results Compared with phosphate bufter saline(PBS)group,combined immunization with HCV-T and HCV-F4HVR1 induced higher levels of specific IgG against HCV-F4HVR1(t=3.815,3.762,P＜0.05),HCV-NS3-specific CTL response(t=3.971,P＜0.05)and IL-4(t=3.512,3.417,P＜0.05)and IFN-γ(t=3.813,3.426,3.671,P＜0.05)secretions.Conclusion High levels of specific humoral immunity and cellular immunity are induced in vivo after combined immunization with HCV-T and HCV-F4 HVR1,which could effectively prevent from the attack of SP2/0-NS3 cells.