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目的观察胸腺素及干扰素基因表达对乙型肝炎基因疫苗pVAX1-S2S的免疫增强效应。方法从乙型肝炎患者血清中扩增S2S基因,构建表达质粒pVAX1-S2S。从成人外周血白细胞总RNA中,逆转录聚合酶链反应扩增干扰素α基因Ⅰ,将其与S2S相连接,构建重组表达质粒pVAX1-I/S2S;将干扰索α基因Ⅰ与人工合成的胸腺素α基因T相连接,构建重组表达质粒pVAX1-T/I。将上述表达质粒分组肌肉接种BALB/c小鼠:单独免疫组接种pVAX1-S2S 100 μg;联合免疫组1:接种pVAX1-I/S2S 100 μg;联合免疫组2:每只小鼠同时接种pVAX1-T/I与pVAX1-S2S各50 μg。上述各组于2、4周后分别加强免疫1次。然后动态检测小鼠血清抗-HBs和前S2抗体。结果接种后3、5、8周,小鼠血清抗-HBs阳转率:联合免疫组1分别为12.5%、12.5%、62.5%;联合免疫组2分别为25%、50%、50%,二者总体上均优于pVAX1- S2S单独免疫组。联合免疫组2的前S2抗体水平则高于其它两组。结论胸腺素α1和(或)干扰素α8 基因的表达具有分子免疫佐剂的效应,能够增强乙型肝炎基因疫苗的特异性体液免疫诱生效力。
Objective To observe the immune enhancement effects of thymosin and interferon gene on hepatitis B gene vaccine pVAX1-S2S. Methods The S2S gene was amplified from the serum of hepatitis B patients and the expression plasmid pVAX1-S2S was constructed. The interferon alpha gene Ⅰ was amplified by reverse transcription polymerase chain reaction (RT-PCR) from total RNA of adult peripheral white blood cells and ligated with S2S to construct the recombinant plasmid pVAX1-I / S2S. Thymosin α gene T phase connection, construct recombinant expression plasmid pVAX1-T / I. BALB / c mice were inoculated intramuscularly with the above expression plasmids: pVAX1-S2S 100 μg was immunized with the group of immunization alone; 100 μg of pVAX1-I / S2S was inoculated with the group of combination immunization 2; pVAX1- 50 μg each of T / I and pVAX1-S2S. The above groups were boosted once after 2,4 weeks. Then mouse anti-HBs and pre-S2 antibodies were detected dynamically. Results The positive rate of anti-HBs in mice serum was 12.5%, 12.5% and 62.5% respectively at 3, 5, and 8 weeks after inoculation. The combined immunization groups were 25%, 50% and 50% Overall, both were superior to the pVAX1-S2S immunized group. Pre-S2 antibody levels in combined immunization group 2 were higher than the other two groups. Conclusion The expression of thymosin α1 and / or interferon α8 has the molecular adjuvant effect and can enhance the specific humoral immunity induced by hepatitis B gene vaccine.