丙型肝炎病毒重组蛋白的免疫保护性

Immunoprotection analysis of hepatitis C virus recombinant protein

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14d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1．0×10^6个SP2／0-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HCVT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SP2／0—NS3细胞的攻击。

Objective To investigate immune responses and protective effect induced by two recombinant proteins of hepatitis C virus （HCV） in BALB/e mice. Methods BALB/c mice were immunized with recombinant proteins HCVT 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 × 10^6 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 hurter 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-specifie 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- F4HVR1, which could effectively prevent from the attack of SP2/0-NS3 cells.