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为探索治疗型HBV基因疫苗治疗慢性乙型肝炎的可行性 ,自行构建了HBV包膜蛋白前S2 ·S基因及人IL 2和IFN γ融合蛋白基因真核表达质粒 ,并将双质粒融合蛋白作为佐剂组合成治疗型HBV基因疫苗 ,在健康小鼠、HBV转基因 (Tg)小鼠、新西兰兔和恒河猴体内进行了免疫效果试验。结果发现 :①特异性CTL活性提高 ;②对HBsAg特异性T细胞增殖能力提高 ,HBsAg 3 0 μg /ml对pcS2 ·S免疫的小鼠脾细胞的刺激指数 (SI =5 6± 0 9)明显较空白质粒pcDNA3 1组 (SI =2 0± 0 5 )为高 (P <0 0 1) ,免疫组的IL 2 /IFN γ分泌水平为 (2 2 6 3± 41 0 5pg/ml) /(5 1 1± 7 7pg/ml) ,明显较空白组的 (69 0± 2 2 1pg/ml) /(0 9± 0 7pg/ml)为高 (P <0 0 1) ;③HBV基因疫苗免疫健康小鼠局部引流淋巴结中DCs的诱导HBsAg致敏T细胞增殖指数 (4 2 0 )较pcDNA3 1组 (2 5 5 )为高 ;⑤用在体电脉冲法注射治疗型HBV基因疫苗后 ,检测血清抗 HBs水平 ,无论是小鼠、兔还是猴均有明显提高。提示该治疗型HBV基因疫苗能较好地诱导体液和细胞免疫 ,为其治疗HBV感染提供了实验依据
In order to explore the feasibility of therapeutic HBV gene vaccine in the treatment of chronic hepatitis B, we constructed the eukaryotic expression plasmid of pre-S2 · S gene of HBV envelope protein and human IL 2 and IFNγ fusion protein, and used the double plasmid fusion protein as Adjuvants were combined to form a therapeutic HBV gene vaccine, and the immune effect test was conducted in healthy mice, HBV transgenic mice (Tg), New Zealand white rabbits and rhesus monkeys. The results showed that: (1) the specific CTL activity was enhanced; (2) the proliferation of HBsAg-specific T cells was enhanced; the stimulation index (SI = 56 ± 0 9) of spleen cells immunized with pcS2 · S by HBsAg 30 μg / The level of IL 2 / IFN γ secretion in the immunized group was (2 2 6 3 ± 41 0 5 pg / ml) / (2) compared with the blank plasmid pcDNA3 1 group (SI = 20 ± 0 5) 5 1 1 ± 7 7pg / ml), which was significantly higher than that of the blank control group (69 0 ± 21 21 pg / ml) / (0 9 ± 0 7 pg / ml) (P 0 01) Induction of DCs in local draining lymph nodes of mice resulted in a higher HBsAg-sensitized T cell proliferation index (420) than that of pcDNA3 1 group (25 5); ⑤ After inoculation of therapeutic HBV gene vaccine by volumetric electrical pulse method, serum Anti-HBs levels, whether mice, rabbits or monkeys have significantly improved. It is suggested that the therapeutic HBV gene vaccine can induce humoral and cellular immunity well and provide an experimental basis for its treatment of HBV infection