Huan Chang-chao, Wang Yue, Ni Bo, Wang Rui, Huang Li, Ren Xiao-feng, Tong Guang-zhi, Ding Chan, Fan Hong-jie, Mao Xiang
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, China.
Arch Virol. 2015 Jul;160(7):1621-8. doi: 10.1007/s00705-015-2408-0. Epub 2015 Apr 22.
It is well known that many viruses use heparan sulfate as the initial attachment factor. In the present study, we determined whether porcine epidemic diarrhea virus (PEDV), an emerging veterinary virus, infects Vero cells by attaching to heparan sulfate. Western blot analysis, real-time PCR, and plaque formation assay revealed that PEDV infection was inhibited when the virus was pretreated with heparin (an analogue of heparan sulfate). There was no inhibitory effect when the cells were pre-incubated with heparin. We next demonstrated that enzymatic removal of the highly sulfated domain of heparan sulfate by heparinase I treatment inhibited PEDV infection. We also confirmed that sodium chlorate, which interferes with heparan sulfate biosynthesis, also inhibited PEDV infection. Furthermore, we examined the effect of two heparin derivatives with different types of sulfation on PEDV infection. The data suggested de-N-sulfated heparin, but not N-acetyl-de-O-sulfated heparin, inhibits PEDV infection. In summary, our studies revealed that heparan sulfate acts as the attachment factor of PEDV in Vero cells.
众所周知，许多病毒利用硫酸乙酰肝素作为初始附着因子。在本研究中，我们确定了一种新出现的兽医病毒——猪流行性腹泻病毒（PEDV）是否通过附着于硫酸乙酰肝素感染Vero细胞。蛋白质免疫印迹分析、实时荧光定量PCR和蚀斑形成试验表明，当病毒用肝素（硫酸乙酰肝素的类似物）预处理时，PEDV感染受到抑制。当细胞用肝素预孵育时，没有抑制作用。接下来，我们证明用肝素酶I处理酶促去除硫酸乙酰肝素的高度硫酸化结构域可抑制PEDV感染。我们还证实，干扰硫酸乙酰肝素生物合成的氯酸钠也抑制PEDV感染。此外，我们研究了两种具有不同硫酸化类型的肝素衍生物对PEDV感染的影响。数据表明，去N-硫酸化肝素而非N-乙酰去O-硫酸化肝素可抑制PEDV感染。总之，我们的研究表明硫酸乙酰肝素在Vero细胞中作为PEDV的附着因子发挥作用。
