摘要
目的 探索细胞因子对丙型肝炎病毒 (HCV)核心(C)基因疫苗诱生免疫应答强度的增强作用 .方法 将包含HCV- C蛋白的基因片段插入真核表达载体 pc DNA3中 ,构建重组质粒 pc DNAHCV- C.将此重组质粒单独或与包含鼠 IL-2或 IL- 12基因的表达质粒共免疫 BAL B/ c小鼠 ,EL ISA法检测 HCV C特异性抗体产生情况 ;以 pc DNAHCV- C转染小鼠 SP2 / 0细胞 ,表达 HCV C抗原者为靶细胞 ,进行 5 1 Cr释放试验 ,检测细胞毒 T淋巴细胞 (CTL s)的特异性杀伤功能 .结果 p IL - 2能够增加 pc DNAHCV- C诱导的特异性抗体的产生 ,而对 CTL的杀伤作用增强不明显 .而 p IL - 12对 pc D-NAHCV- C诱导的抗体和细胞免疫应答均有增强 ,尤以 CTL的杀伤作用增强为显著 (P<0 .0 5 ) .结论 佐以不同的细胞因子 ,可增强基因疫苗诱生的免疫应答强度 ,可能使机体对基因疫苗的免疫应答朝着有利于优先诱导 CTL 免疫应答 ,清除病原体的方向发展 .
AIM To investigate the enhancing effect of IL-2 or IL-12 on immune responses of BALB/c mice to HCV C gene. METHODS BALB/c mice were immunized with a recombinant plasmid containing HCV-C gene (pcDNAHCV-C) alone or together with a constructs encoding IL-2 or IL-12(pIL-2 or pIL-12). Anti-HCV-C specific antibody in the sera was measured with ELISA. Spleen cells from experimental and control mice were extracted. The mouse myloma SP2/0 cells expressing HCV C antigen served as target cells. CTL activity was measured by standard 4-h 51 Cr release assays. RESULTS Anti-HCV-C antibody OD value from 1.21 of pcDNAHCV-C alone increased to 2.09 and 1.73 of coimmunized with pIL-2 or pIL-12 respectively. CTL activity to HCV core protein was enhanced substantially after coimmunization with the IL-12 expression plasmid. In contrast, the improvement of HCV core-specific CTL activity with an IL-2-producing construct coimmunization was not obvious, which maybe resulted from the low-level expression of IL-2 vector-pDOR. CONCLUSION DNA vaccine of HCV combined with IL-12 expressive plasmid can get better therapeutic effects, and it might represent an approach to the gene therapy against HCV infection.
出处
《第四军医大学学报》
2000年第7期834-837,共4页
Journal of the Fourth Military Medical University
