DNA-vaccine platform development against H1N1 subtype of swine influenza A viruses.
DNA-vaccine platform development against H1N1 subtype of swine influenza A viruses.
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DOI：
10.1089/vim.2011.0093
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发表时间：
2012-08
期刊：
影响因子：
2.2
通讯作者：
Huiling Wei;S. Lenz;D. H. Thompson;R. Pogranichniy
中科院分区：
文献类型：
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作者：
Huiling Wei;S. Lenz;D. H. Thompson;R. Pogranichniy
猪流感病毒（SIV）是猪群中一种重要的病毒性病原体。然而，商业疫苗不能仅通过诱导体液免疫提供完全保护，并且需要经常更新以对抗当前的分离株。DNA疫苗接种是一种有效的手段，引发免疫系统的两臂，体液和细胞免疫反应。本研究通过在CMV启动子区下游插入嵌合内含子，并通过Kozak序列插入DNA载体pcDNA3.1，以增强插入基因的表达。编码牛疱疹病毒-1被膜蛋白的VP22基因（VP22c）的c端分别与四个四复制表位的n端融合：两个b细胞表位（HA91-108和M2e）和两个t细胞表位（NP366-374和NP380-393），至少在北美流行的三种SIV亚型猪群体中是保守的。linkker - kk -用于间隔两个b细胞表位的每个拷贝，- rvkr -用于两个t细胞表位，以增强表位向免疫系统的呈递。体外转染293FT细胞证实了表位的表达，含有VP22c的质粒比不含VP22c的质粒获得了更高的表位阳性细胞百分比。将DNA质粒联合或单独给予小鼠肌内注射，或与VP22c融合的四复制表位重组蛋白增强后，疫苗刺激了对两个b细胞表位的所需表位特异性体液免疫，以及对表位NP380-393的细胞免疫。我们的研究结果表明，与VP22c融合的具有四复制表位的质粒可能是开发SIV表位作为疫苗的潜在载体。
Swine influenza virus (SIV) is an important viral pathogen in pig populations. However, commercial vaccines cannot provide complete protection with induced humoral immunity only, and require frequent updates to fight against current isolates. DNA vaccination is an effective means of eliciting both arms of the immune system, the humoral and cellular immune responses. In this study, DNA vector pcDNA3.1 was inserted with a chimeric intron downstream of the CMV promoter region followed by a Kozak sequence to enhance the expression of gene inserts. The C-terminal of the VP22 gene (VP22c), encoding the tegument protein of bovine herpesvirus-1, was fused separately to the N-terminal of four quadruplicated epitopes: two B-cell epitopes (HA91-108 and M2e), and two T-cell epitopes (NP366-374 and NP380-393), which were conserved, at least among the three SIV subtypes prevailing in pig populations in North America. Linker -KK- was used to space between each copy of the two B-cell epitopes, and -RVKR- was used for the two T-cell epitopes, in order to enhance the presentation of epitopes to the immune system. The expression of epitopes was confirmed in in vitro transfection of 293FT cells, and higher percentages of epitope-positive cells were achieved from the plasmids containing VP22c than those without. After the DNA plasmids were administered to mice intramuscularly in combination or separately, or boosted with recombinant proteins of quadruplicated epitopes fused to VP22c, the vaccine stimulated the desired epitope-specific humoral immunity to the two B-cell epitopes, and cellular immunity to the epitope NP380-393. Our results indicate that plasmids with quadruplicated epitopes fused to the VP22c may be a potential vehicle in developing epitopes as vaccines against SIV.