携带抑胃多肽序列的HBc 病毒样颗粒蛋白疫苗的制备及其免疫效果
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国家“十一五”科技支撑课题(2006BAI02B08).


Preparation of HBc virus-like particle vaccine harboring sequences of gastric inhibitory peptide and its immune effects
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Supported by National Key Technology R&D Program(2006BAI02B08).

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    摘要:

    目的:设计及制备携带抑胃多肽(GIP)抗原序列的HBc病毒样颗粒(VLP)蛋白疫苗,并研究其免疫效果,为肥胖免疫干预治疗提供新的策略。方法:克隆GIP cDNA序列,并与HBc VLP(1~144aa)cDNA序列进行融合,通过原核表达及纯化,制备携带GIP序列的HBc VLP疫苗(HBc-GIP);同时将该融合基因的cDNA序列克隆入pVAX1真核表达载体,制备GIP核酸疫苗。用2种疫苗免疫大鼠并进行免疫学指标评价。结果:成功制备了携带GIP抗原序列的HBc-GIP,并同时制备了编码该融合蛋白序列的核酸疫苗pVAX1-HBc-GIP。经过两种疫苗的联合免疫,可以获得高滴度的GIP特异性抗体IgG,显示了良好的免疫效果。结论:采用HBc(1~144aa)作为GIP的载体,制备VLP蛋白疫苗,联合核酸疫苗免疫可以有效地诱导靶向大鼠GIP的特异性体液免疫反应,打破大鼠自身抗原耐受。该疫苗可以作为一种新的控制肥胖的免疫干预药物来进一步深入研究。

    Abstract:

    Objective:To design and prepare HBc virus-like particle vaccine (VLP) harboring the specified epitope of gastric inhibitory peptide (GIP),and to study its immune effects,so as to cast new lights on immune therapy of obesity.Methods: GIP cDNA was cloned and was fused with HBc VLP (1-144aa) cDNA.Through prokaryotic expression and purification processes,a GIP vaccine linked to the HBc-derived VLP carrier was constructed.We also cloned the fusion gene into eukaryotic vector pVAX1 to prepare GIP nuclear acid vaccine.And the two kinds of vaccines were tested in rats for their immunological parameters.Results: GIP vaccine linked to HBc-derived VLP carrier was successfully prepared.We also prepared the genetic vaccine pVAX1-HBc-GIP.The combined vaccination with both vaccines could obtain high titer and peptide-specific IgG antibody,displaying a satisfactory immunogenicity.Conclusion: GIP protein vaccine using HBc as immune-enhancing vaccine carrier can effectively induce GIP-specific humoral immunity,disrupting the tolerance to self antigens in rats.The vaccine is worth studying as a new drug for control of obesity.

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  • 收稿日期:2008-12-30
  • 最后修改日期:2009-04-30
  • 录用日期:2009-08-27
  • 在线发布日期: 2009-11-18
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