Identification of a novel cell culture adaptation site on the capsid of foot-and-mouth disease virus.
Identification of a novel cell culture adaptation site on the capsid of foot-and-mouth disease virus.
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DOI：
10.1099/jgv.0.000222
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发表时间：
2015-09
期刊：
影响因子：
--
通讯作者：
Jackson T
中科院分区：
文献类型：
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作者：
Chamberlain K;Fowler VL;Barnett PV;Gold S;Wadsworth J;Knowles NJ;Jackson T
疫苗接种仍然是流行国家控制口蹄疫的最有效工具，也是预防新疫情的应急准备。口蹄疫疫苗是化学灭活的病毒制剂，新疫苗的生产严重依赖于野外病毒的细胞培养适应性，这可能证明是有问题的。细胞培养适应的主要驱动力是受体可用性。口蹄疫病毒（FMDV）的田间分离株使用RGD依赖性整合素作为受体，而细胞培养适应通常选择具有改变的受体偏好的变体。以前，已经鉴定了衣壳上的两个独立位点，其中突变与改善的细胞培养物生长相关。一种是由三种主要结构蛋白（VP 1-VP 3）形成的浅凹陷，其中突变产生硫酸乙酰肝素（HS）结合位点（典型的HS结合位点）。另一个涉及VP 1的残基，位于五重对称轴上。对于一些病毒，在这个网站的变化导致HS结合;对于其他人，受体是未知的。在这里，我们报告了VP 2上一个新位点的鉴定，该位点的突变导致A/IRN/87疫苗变体（称为A-）的细胞嗜性扩大。此外，我们发现，将相同的突变引入不同的A型野外病毒（A/TUR/2/2006）中，导致与A/IRN/87 A −疫苗变体相同的扩增细胞培养嗜性。这些观察结果增加了FMDV的多细胞附着机制的证据，并且当细胞培养适应证明困难时，可能对疫苗制造有用。
Vaccination remains the most effective tool for control of foot-and-mouth disease both in endemic countries and as an emergency preparedness for new outbreaks. Foot-and-mouth disease vaccines are chemically inactivated virus preparations and the production of new vaccines is critically dependent upon cell culture adaptation of field viruses, which can prove problematic. A major driver of cell culture adaptation is receptor availability. Field isolates of foot-and-mouth disease virus (FMDV) use RGD-dependent integrins as receptors, whereas cell culture adaptation often selects for variants with altered receptor preferences. Previously, two independent sites on the capsid have been identified where mutations are associated with improved cell culture growth. One is a shallow depression formed by the three major structural proteins (VP1–VP3) where mutations create a heparan sulphate (HS)-binding site (the canonical HS-binding site). The other involves residues of VP1 and is located at the fivefold symmetry axis. For some viruses, changes at this site result in HS binding; for others, the receptors are unknown. Here, we report the identification of a novel site on VP2 where mutations resulted in an expanded cell tropism of a vaccine variant of A/IRN/87 (called A − ). Furthermore, we show that introducing the same mutations into a different type A field virus (A/TUR/2/2006) resulted in the same expanded cell culture tropism as the A/IRN/87 A − vaccine variant. These observations add to the evidence for multiple cell attachment mechanisms for FMDV and may be useful for vaccine manufacture when cell culture adaptation proves difficult.