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首页> 外文期刊>World Journal of Gastroenterology >Construction of exogenous multiple epitopes of helper T Iymphocytes and DNA immunization of its chimeric plasmid with HBV pre-S2/S gene
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Construction of exogenous multiple epitopes of helper T Iymphocytes and DNA immunization of its chimeric plasmid with HBV pre-S2/S gene

机译：辅助T淋巴细胞外源多表位的构建及其HBV pre-S2 / S基因嵌合质粒的DNA免疫

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ABM: To design and construct, an exogenous multiple epitope of helper T lymphocytes (HTL), and to evaluate its effect on anti-HBs response through DNA immunization. METHODS: Artificial HTL epitope, PADRE and four other HTL epitopes from different proteins were linked together using splicing by overlap extension to generate exogenous multiple epitopes of HTL, MTE5. pcMTE5 and pcHB were generated by cloning MTE5 and fragments of HBV pre-S2/S gene into mammalian expression plasmid pcDNA3. Four chimeric plasmids were constructed by cloning MTE5 into the region of pre-S2 gene (Bam HI), 5′ terminal of S gene (HincII, Xba I) and 3′ terminal of S gene (Acc I) of pcHB respectively. BALB/c mice were used in DNA immunization of the recombinant plasmids. Anti-HBs was detected using Abbott IMx AUSAB test kits. RESULTS: The sequences of MTE5 and the 6 constructs of recombinant plasmids were confirmed to be correct by DNA sequencing. The anti-HBs response of the co-inoculation of pcHB and pcMTE5 was much higher than that of the inoculation of pcHB only (136.7 +- 69.1 mIU/mL vs 27.6 +- 17.3 mIU/mL, P < 0.01, t = -6.56). Among the 4 chimeric plasmids, only the plasmid in which MTE5 was inserted into the pre-S2 region had good anti-HBs response (57.54 +- 7.68 mIU/mL), and had no significant difference compared with those of pcHB and the co-inoculation of pcHB and pcMTE5. CONCLUSION: Exogenous multiple epitopes of HTL had immune enhancement when they were co-inoculated with pre-S2/S gene or inoculated in the chimeric form at a proper site of pre-S2/S gene of HBV. It might suggest that it was possible to improve hepatitis B vaccine using exogenous multiple epitopes of HTL. The antibody responses were very low using DNA immunization in the study. Thus, the immune enhancement effect of exogenous multiple epitopes of HTL has to be confirmed and the effect on overcoming the drawback of the polymorphism of HLA Ⅱ antigens should also be evaluated after these chimeric plasmids are expressed in mammalian cell lines.
机译：ABM：设计和构建辅助T淋巴细胞（HTL）的外源多个表位，并通过DNA免疫评估其对抗HBs反应的作用。方法：通过重叠延伸拼接，将来自不同蛋白质的人工HTL表位，PADRE和其他四个HTL表位连接在一起，以生成HTL，MTE5的外源多个表位。通过将MTE5和HBV pre-S2 / S基因的片段克隆到哺乳动物表达质粒pcDNA3中来生成pcMTE5和pcHB。通过将MTE5分别克隆到pcHB的pre-S2基因（Bam HI），S基因的5'末端（HincII，Xba I）和S基因的3'末端（Acc I）区域，构建了四个嵌合质粒。 BALB / c小鼠用于重组质粒的DNA免疫。使用雅培IMx AUSAB测试套件检测到抗HBs。结果：通过DNA测序证实了MTE5的序列和6个重组质粒的构建是正确的。联合接种pcHB和pcMTE5的抗HBs应答比仅接种pcHB的要高得多（136.7±69.1 mIU / mL vs 27.6±17.3 mIU / mL，P <0.01，t = -6.56 ）。在这4种嵌合质粒中，只有将MTE5插入pre-S2区域的质粒具有良好的抗HBs反应（57.54±7.68 mIU / mL），与pcHB和co-接种pcHB和pcMTE5。结论：HTL的外源多个表位与前S2 / S基因共同接种或以嵌合形式接种在HBV前S2 / S基因的适当位点具有免疫增强作用。这可能表明有可能使用外源的HTL多表位来改善乙型肝炎疫苗。在研究中使用DNA免疫的抗体反应非常低。因此，在哺乳动物细胞系中表达这些嵌合质粒后，必须确定HTL的外源多个表位的免疫增强作用，并且还应评估克服HLAⅡ抗原多态性缺陷的作用。

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