重组体pCD-HCV1诱发小鼠免疫反应的研究

Study on immune responses to recombinant protein in mice inoculated with pCD HCV1 DNA recombinant

目的探讨丙型肝炎病毒（ＨＣＶ）基因重组体诱发小鼠免疫反应。方法将编码Ⅱ型ＨＣＶ结构蛋白的基因片段Ｃ、Ｅ１和大部分Ｅ２插入到真核细胞表达载体ｐＣＤＳＲα１中，构建成重组体ｐＣＤＨＣＶ１。经肌内注射将此重组体免疫Ｂａｌｂ／ｃ小鼠。结果ｐＣＤＨＣＶ１（１００μｇ／只，ｎ＝１２）３～４次接种小鼠后，血清抗体水平达０．７１～０．７７（Ａ值，下同），空载体ｐＣＤＳＲα１免疫鼠（ｎ＝８），血清抗体阴性；对其中８只免疫鼠持续检测１８周，未见抗体水平有下降趋势（０．６８～０．７５）。重组体ｐＣＤＨＣＶ１免疫鼠（ｎ＝１２）的脾细胞对ＨＣＶ合成肽ＣＰ９、基因重组抗原Ｃ、Ｅ１刺激后，均出现增殖反应（ｃｐｍ），ＳＩ分别为４．１４±０．９，３．９８±１．６，４．０２±１．２，与ｐＣＤＳＲα１免疫鼠（ｎ＝８）相比，差异有显著性（Ｐ＜０．００１）。结论构建的ＨＣＶＤＮＡ重组体可诱发Ｂａｌｂ／ｃ小鼠产生免疫反应。

Objective To inquire into the immune responses to expression protein in mice immunized with pCD HCV1 recombinant.Methods The gene fragment encoding C and HCV Ⅱ type was inserted into pCD SRα1 of expression vector and formed pCD HCV1 and then was injected into quadriceps muscles of Balb/c mice. Results The serum antibody level reached to 0.71～0.77(A value, the same below) after recombinant pCD HCV1(100μg/mouse) were inoculated into mice( n =12) three times or four times and blank vector pCD SRα1 could not induce mice( n =8) to generate antibody in the same way. After the antibody levels in mice( n =8) immunized by pCD HCV1 ascended to peak value(0.71), it did not occur to be the trend of descending during 18 weeks detected (0.68～0.75). The spleen cells prolitferative responses to HCV synthetic peptide CP9 and gene recombinant antigens C and E1 in mice immunized with pCD HCV1 were detected and its stimulation index (SI) were 4.14±0.9, 3.98±1.6 and 4.02±1.2 respectively and there was a significant difference ( P <0.001) comparing with that of spleen cells of mice immunized with pCD SRα1.Conclusion The investigations demonstrated that recombinant pCD HCV1 can elicite Blab/c mice to generate antibody and spleen cells proliferative responses to HCV antigens via DNA immunization.