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目的探讨ＨＣＶ复合多表位ＤＮＡ疫苗的可行性。方法把ＨＣＶ多表位抗原基因ＰＣＸ克隆到真核表达载体ｐＲＥＰ９（ＲＳＶ启动子）及ｐＣＤＮＡ３（ＣＭＶ启动子）中，构建真核表达载体ｐＲＥＰ９／ＰＣＸ及ｐｃＤＮＡ３／ＰＣＸ。将其肌肉注射免疫小鼠及家兔，检测特异性体液免疫和细胞免疫的水平，并观察免疫小鼠的安全性。结果质粒ｐＲＥＰ９／ＰＣＸ及ｐｃＤＮＡ３／ＰＣＸ于免疫后第 ６ｗｋ和第 １０ ｗｋ时，开始可检测到抗 ＧＺ－ＰＣＸ ＩｇＧ，随后抗体滴度逐渐升高，但水平较低，持续时间较短。ｐｃＤＮＡ３／ＰＣＸ肌肉注射免疫家兔后，于第８ｗｋ开始出现特异性抗体，至 ３个月后滴度升至 １： ３ ２００，随后也开始下降。免疫小鼠及家兔可诱发针对ＧＺ－ＰＣＸ融合蛋白的迟发型超敏反应，并刺激淋巴细胞转化。免疫后小鼠的体重正常，肝脾脏未见明显肿大，具有良好的安全性。结论ＨＣＶ多表位基因可诱发特异性免疫应答且安全性好，为ＨＣＶ疫苗的研究提供了一定的理论及实验依据。
Objective To investigate the feasibility of HCV multi-epitope DNA vaccine. Methods PCX gene of HCV multi-epitope antigen was cloned into eukaryotic expression vector pREP9 (RSV promoter) and pCDNA3 (CMV promoter), and eukaryotic expression vector pREP9 / PCX and pcDNA3 / PCX were constructed. The mice were injected intramuscularly with rabbits and rabbits to detect the level of specific humoral and cellular immunity, and the safety of immunized mice was observed. Results The anti-GZ-PCX IgG was detected at the 6th week and the 10th week after the immunization with plasmid pREP9 / PCX and pcDNA3 / PCX. Then the titer of antibody was gradually increased but the level was lower and the duration was shorter. After immunization of rabbit with pcDNA3 / PCX intramuscular injection, specific antibodies started to appear on the 8th week, and the titer rose to 1: 3 200 after 3 months, and then began to decline. Immunized mice and rabbits induced delayed-type hypersensitivity to GZ-PCX fusion protein and stimulated lymphocyte transformation. After immunization, the mice had normal body weight, no obvious enlargement in liver and spleen, and good safety. Conclusion The HCV multi-epitope gene can induce specific immune response with good safety and provide some theoretical and experimental evidences for the study of HCV vaccine.