Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development.
Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development.
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DOI：
10.1038/s41598-022-26766-y
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发表时间：
2023-01-06
影响因子：
4.6
通讯作者：
中科院分区：
文献类型：
--
作者：
目前针对牛病毒性腹泻病毒(BVDV)的活疫苗、减毒疫苗和灭活疫苗都有其局限性。在这里，我们报道了BVDV亚单位疫苗的开发，方法是(I)利用BHK21细胞表达分泌形式的重组E2糖蛋白，(Ii)测定小鼠的免疫反应。通过删除C端跨膜锚定结构域并将其融合到V5表位标签来修饰E2糖蛋白。这使得可以使用抗V5单抗进行检测，并简单地从细胞培养液中纯化表达的、分泌的E2形式。此外，我们通过基因融合的方式将绿色荧光蛋白(GFP)与E2结合在一起，从而产生了一个自我处理的多蛋白[GFP-T2A-BVDV-E2Trunk-V5]，产生了离散的[GFP-T2A]和[E2Trunk-V5]翻译产物：GFP荧光作用。因此，作为E2表达的替代标记，从细胞培养上清液中纯化的[E2-V5]接种BALB/c小鼠，观察到了体液和细胞免疫反应。因此，我们的抗原表达系统提供了(I)简单的抗原纯化方案和(Ii)进一步大规模生产疫苗的可行策略。
Both current live, attenuated, and killed virus vaccines for bovine viral diarrhea virus (BVDV) have their limitations. Here, we report the development of a BVDV subunit vaccine by (i) the expression of a secreted form of a recombinant E2 glycoprotein using BHK21 cells and (ii) determination of the immune responses in mice. The E2 glycoprotein was modified by deletion of the C-terminal transmembrane anchor domain and fusion to a V5 epitope tag. This allowed detection using anti-V5 monoclonal antibodies together with simple purification of the expressed, secreted, form of E2 from the cell media. Furthermore, we genetically fused green fluorescent protein (GFP) linked to E2 via a Thosea asigna virus 2A (T2A) ribosome skipping sequence thereby creating a self-processing polyprotein [GFP-T2A-BVDV-E2trunk-V5], producing discrete [GFP-T2A] and [E2trunk-V5] translation products: GFP fluorescence acts, therefore, as a surrogate marker of E2 expression, BALB/c mice were inoculated with [E2trunk-V5] purified from cell media and both humoral and cellular immune responses were observed. Our antigen expression system provides, therefore, both (i) a simple antigen purification protocol together with (ii) a feasible strategy for further, large-scale, production of vaccines.
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影响因子：
3.5
作者：
Minskaia E;Nicholson J;Ryan MD
通讯作者：
Ryan MD
影响因子：
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Buonavoglia C
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de Felipe, P;Ryan, MD
通讯作者：
Ryan, MD
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通讯作者：
Kovarcik, K.