Murakami Shin, Takenaka-Uema Akiko, Kobayashi Tomoya, Kato Kentaro, Shimojima Masayuki, Palmarini Massimo, Horimoto Taisuke
Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan
Department of Infection Control and Disease Prevention, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
J Virol. 2017 Jul 12;91(15). doi: 10.1128/JVI.00503-17. Print 2017 Aug 1.
(AKAV) and Schmallenberg virus (SBV) are members of the genus , which are transmitted by arthropod vectors with a broad cellular tropism as well as Both AKAV and SBV cause arthrogryposis-hydranencephaly syndrome in ruminants. The main cellular receptor and attachment factor for entry of these orthobunyaviruses are unknown. Here, we found that AKAV and SBV infections were inhibited by the addition of heparin or enzymatic removal of cell surface heparan sulfates. To confirm this finding, we prepared heparan sulfate proteoglycan (HSPG)-knockout (KO) cells by using a clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 system and measured the quantities of binding of these viruses to cell surfaces. We observed a substantial reduction in AKAV and SBV binding to cells, limiting the infections by these viruses. These data demonstrate that HSPGs are important cellular attachment factors for AKAV and SBV, at least , to promote virus replication in susceptible cells. AKAV and SBV are the etiological agents of arthrogryposis-hydranencephaly syndrome in ruminants, which causes considerable economic losses in the livestock industry. Here, we identified heparan sulfate proteoglycan as a major cellular attachment factor for the entry of AKAV and SBV. Moreover, we found that heparin is a strong inhibitor of AKAV and SBV infections. Revealing the molecular mechanisms of virus-host interactions is critical in order to understand virus biology and develop novel live attenuated vaccines.
阿卡斑病毒（AKAV）和施马伦贝格病毒（SBV）是该属的成员，它们由具有广泛细胞嗜性的节肢动物媒介传播，并且AKAV和SBV都会在反刍动物中引起关节弯曲-积水性无脑综合征。这些正布尼亚病毒进入细胞的主要细胞受体和附着因子尚不清楚。在这里，我们发现添加肝素或酶促去除细胞表面硫酸乙酰肝素可抑制AKAV和SBV感染。为了证实这一发现，我们使用成簇规律间隔短回文重复序列（CRISPR）-Cas9系统制备了硫酸乙酰肝素蛋白聚糖（HSPG）敲除（KO）细胞，并测量了这些病毒与细胞表面结合的量。我们观察到AKAV和SBV与细胞的结合大幅减少，限制了这些病毒的感染。这些数据表明，HSPG至少是AKAV和SBV重要的细胞附着因子，可促进病毒在易感细胞中复制。AKAV和SBV是反刍动物关节弯曲-积水性无脑综合征的病原体，给畜牧业造成了相当大的经济损失。在这里，我们确定硫酸乙酰肝素蛋白聚糖是AKAV和SBV进入细胞的主要细胞附着因子。此外，我们发现肝素是AKAV和SBV感染的强力抑制剂。揭示病毒-宿主相互作用的分子机制对于理解病毒生物学和开发新型减毒活疫苗至关重要。
