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构建并免疫编码鼠补体 C3d-p28 融合蛋白和猪繁殖与呼吸综合征病毒 GP5 基因的 DNA 疫苗。

Construction and immunogenicity of DNA vaccines encoding fusion protein of murine complement C3d-p28 and GP5 gene of porcine reproductive and respiratory syndrome virus.

机构信息

College of Veterinary Medicine, Shandong Agricultural University, Taian, Shandong 271018, China.

出版信息

Vaccine. 2011 Jan 17;29(4):629-35. doi: 10.1016/j.vaccine.2010.11.046. Epub 2010 Dec 4.

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) has recently caused catastrophic losses in swine industry worldwide. The commercial vaccines only provide a limited protection against PRRSV infection. At present, DNA vaccine is the focus on the new vaccines. The gene fragment (p28) coding for the molecular adjuvants complement protein C3d (mC3d) from BALB/c mouse was cloned and expressed as a fusion protein for its application in the vaccine study of mice. Three potential vaccines construct units were engineered to contain two, four and six copies of mC3d-p28 coding gene linked to the GP5 gene of PRRSV and one vaccine expressing GP5 alone (pcDNA3.1-GP5) was constructed. Subsequently, the vaccines' abilities to elicit the humoral and cellular immune responses were investigated in mice. These results showed that significantly enhanced GP5-specific ELISA antibody, GP5-specific neutralizing antibody, IFN-γ level, and IL-4 level, could be induced in mice immunized with DNA construct units encoding the pcDNA3.1-C3d-p28.n-GP5 than those received DNA vaccine expressing GP5 alone (pcDNA3.1-GP5). Analysis of the immunogenicity of different repeats of mC3d-p28 revealed that mC3d-p28 had an enhancing effect on the immunogenicity of antigens, and that six or more repeats of mC3d-p28 may be necessary for efficient enhancement of antigen specific immune responses. This approach may provide a new strategy for the development of efficient vaccines against the PRRSV for pigs in the future.

摘要

猪繁殖与呼吸综合征病毒(PRRSV)最近在全球范围内给养猪业造成了灾难性的损失。商业疫苗只能对 PRRSV 感染提供有限的保护。目前,DNA 疫苗是新型疫苗的重点。该研究从 BALB/c 小鼠中克隆并表达了编码分子佐剂补体蛋白 C3d(mC3d)的基因片段(p28),作为融合蛋白用于研究其在小鼠疫苗中的应用。设计了三个潜在的疫苗构建单元,分别包含两个、四个和六个拷贝的 mC3d-p28 编码基因与 PRRSV 的 GP5 基因相连,以及一个单独表达 GP5 的疫苗(pcDNA3.1-GP5)。随后,在小鼠中研究了这些疫苗引发体液和细胞免疫应答的能力。结果表明,与单独接受表达 GP5 的 DNA 疫苗(pcDNA3.1-GP5)的小鼠相比,接受编码 pcDNA3.1-C3d-p28.n-GP5 的 DNA 构建单元免疫的小鼠可诱导出显著增强的 GP5 特异性 ELISA 抗体、GP5 特异性中和抗体、IFN-γ 水平和 IL-4 水平。对 mC3d-p28 不同重复次数的免疫原性分析表明,mC3d-p28 对抗原的免疫原性具有增强作用,并且 mC3d-p28 重复六次或更多次可能是增强抗原特异性免疫应答的必要条件。该方法可为未来开发针对 PRRSV 的高效猪用疫苗提供新策略。

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