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Virol J. 2019 Nov 12;16(1):134. doi: 10.1186/s12985-019-1246-1.

Heparin sulfate is the attachment factor of duck Tembus virus on both BHK21 and DEF cells.

Wu S1, Wu Z1, Wu Y1, Wang T1, Wang M1,2,3, Jia R1,2,3, Zhu D1,2,3, Liu M1,2,3, Zhao X1,2,3, Yang Q1,2,3, Wu Y1,2,3, Zhang S1,2,3, Liu Y1, Zhang L1, Yu Y1, Pan L1, Chen S4,5,6, Cheng A7,8,9.

Author information

1
Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang District, Chengdu City, 611130, Sichuan Province, China.
2
Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, 611130, Sichuan Province, China.
3
Key Laboratory of Animal Disease and Human Health of Sichuan Province, Wenjiang District, Chengdu City, 611130, Sichuan Province, China.
4
Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang District, Chengdu City, 611130, Sichuan Province, China. shunchen@sicau.edu.cn.
5
Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, 611130, Sichuan Province, China. shunchen@sicau.edu.cn.
6
Key Laboratory of Animal Disease and Human Health of Sichuan Province, Wenjiang District, Chengdu City, 611130, Sichuan Province, China. shunchen@sicau.edu.cn.
7
Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang District, Chengdu City, 611130, Sichuan Province, China. chenganchun@vip.163.com.
8
Institute of Preventive Veterinary Medicine, College of Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, 611130, Sichuan Province, China. chenganchun@vip.163.com.
9
Key Laboratory of Animal Disease and Human Health of Sichuan Province, Wenjiang District, Chengdu City, 611130, Sichuan Province, China. chenganchun@vip.163.com.

Abstract

BACKGROUND:

Duck tembusu virus (DTMUV, genus Flaviviruses, family Flaviviridae) is an emerging flavivirus that can infect a wide range of cells and cell lines in vitro, though the initial step of virus invasion remains obscure.

METHODS:

In this study, drug treatments that including heparin, chondroitin sulfate, heparinase I, chondroitinase ABC and trypsin were applied to detect the influence of DTMUV absorption, subsequently, the copy number of viral genome RNA was analyzed by quantitative real-time PCR. The inhibition process of viral absorption or entry by heparin was determined by western blotting, and the cytotoxicity of drug treated cells was detected by cell counting kit-8.

RESULTS:

We found that the desulfation of glycosaminoglycans (GAGs) with sodium chlorate had a significant effect on the adsorption of DTMUV in both BHK21 and DEF cells. Based on this result, we incubated cells with a mixture of DTMUV and GAGs competition inhibitors or pre-treated cells with inhibitors, after incubation with the virus, the NS5 expression of DTMUV and viral titers were detected. The data suggested that heparin can significantly inhibit the absorption of DTMUV in a dose dependent manner but not at the step of viral entry in BHK21 and DEF cells. Meanwhile, heparinase I can significantly inhibit DTMUV attachment step.

CONCLUSIONS:

Our results clearly proved that heparin sulfate plays an important role in the first step of DTMUV entry, viral attachment, in both BHK21 and DEF cells, which sheds light on the entry mechanism of DTMUV.

KEYWORDS:

Attachment factor; Chondroitin sulfate; Entry; Glycosaminoglycans; Heparin sulfate

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