Department of Genetic Engineering, Faculty of Life Sciences and Technology, Sungkyunkwan University, Suwon 440-746, Korea.
Immune Netw. 2009 Dec;9(6):265-73. doi: 10.4110/in.2009.9.6.265. Epub 2009 Dec 31.
Foot-and-mouth disease virus (FMDV) is a small single-stranded RNA virus which belongs to the family Picornaviridae, genus Apthovirus. It is a principal cause of FMD which is highly contagious in livestock. In a wild type virus infection, infected animals usually elicit antibodies against structural and non-structural protein of FMDV. A structural protein, VP1, is involved in neutralization of virus particle, and has both B and T cell epitopes. A RNA-dependent RNA polymerase, 3D, is highly conserved among other serotypes and strongly immunogenic, therefore, we selected VP1 and 3D as vaccine targets.
VP1 and 3D genes were codon-optimized to enhance protein expression level and cloned into mammalian expression vector. To produce recombinant protein, VP1 and 3D genes were also cloned into pET vector. The VP1 and 3D DNA or proteins were co-immunized into 5 weeks old BALB/C mice.
Antigen-specific serum antibody (Ab) responses were detected by Ab ELISA. Cellular immune response against VP1 and 3D was confirmed by ELISpot assay.
The results showed that all DNA- and protein-immunized groups induced cellular immune responses, suggesting that both DNA and recombinant protein vaccine administration efficiently induced Ag-specific humoral and cellular immune responses.
口蹄疫病毒（FMDV）是一种小的单链 RNA 病毒，属于小 RNA 病毒科，口疮病毒属。它是引起口蹄疫的主要原因，在牲畜中具有高度传染性。在野生型病毒感染中，受感染的动物通常会针对 FMDV 的结构蛋白和非结构蛋白产生抗体。结构蛋白 VP1 参与病毒颗粒的中和，具有 B 细胞和 T 细胞表位。RNA 依赖的 RNA 聚合酶 3D 在其他血清型之间高度保守且具有强烈的免疫原性，因此我们选择 VP1 和 3D 作为疫苗靶标。
为了提高蛋白表达水平，对 VP1 和 3D 基因进行了密码子优化，并将其克隆到哺乳动物表达载体中。为了生产重组蛋白，还将 VP1 和 3D 基因克隆到 pET 载体中。将 VP1 和 3D DNA 或蛋白共同免疫 5 周龄 BALB/C 小鼠。
通过 Ab ELISA 检测抗原特异性血清抗体（Ab）反应。通过 ELISpot 测定法证实了针对 VP1 和 3D 的细胞免疫反应。
结果表明，所有 DNA 和蛋白免疫组均诱导了细胞免疫反应，表明 DNA 和重组蛋白疫苗的联合接种能有效诱导 Ag 特异性体液和细胞免疫反应。
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