Cell surface α2,3-linked sialic acid facilitates Zika virus internalization

Chee Wah Tan,Catherine Hong Huan Hor,Swee Sen Kwek,Han Kang Tee,I-Ching Sam,Eyleen L K Goh,Eng Eong Ooi,Yoke Fun Chan,Lin-Fa Wang

doi:10.1080/22221751.2019.1590130

Chee Wah Tan, Catherine Hong Huan Hor + Show 7 more

Open Access

https://doi.org/10.1080/22221751.2019.1590130

Copy DOI

Abstract

ABSTRACT The emergence of neurotropic Zika virus (ZIKV) raised a public health emergency of global concern. ZIKV can cross the placental barrier and infect foetal brains, resulting in microcephaly, but the pathogenesis of ZIKV is poorly understood. With recent findings reporting AXL as a type I interferon antagonist rather than an entry receptor, the exact entry mechanism remains unresolved. Here we report that cell surface sialic acid plays an important role in ZIKV infection. Removal of cell surface sialic acid by neuraminidase significantly abolished ZIKV infection in Vero cells and human induced-pluripotent stem cells-derived neural progenitor cells. Furthermore, knockout of the sialic acid biosynthesis gene encoding UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase resulted in significantly less ZIKV infection of both African and Asian lineages. Huh7 cells deficient in α2,3-linked sialic acid through knockout of ST3 β-galactoside-α2,3-sialyltransferase 4 had significantly reduced ZIKV infection. Removal of membrane-bound, un-internalized virus with pronase treatment revealed the role of sialic acid in ZIKV internalization but not attachment. Sialyllactose inhibition studies showed that there is no direct interaction between sialic acid and ZIKV, implying that sialic acid could be mediating ZIKV-receptor complex internalization. Identification of α2,3-linked sialic acid as an important host factor for ZIKV internalization provides new insight into ZIKV infection and pathogenesis.

Highlights

Zika virus (ZIKV) is an arthropod-borne RNA virus in the genus Flavivirus with other vector-borne viruses significant to human health, such as dengue virus (DENV), yellow fever virus (YFV), West Nile virus (WNV), and Japanese encephalitis virus (JEV) [1]
We provide evidence that cell surface sialic acid facilitates ZIKV infection in Vero, Huh7, and induced-pluripotent stem cells-derived human neural progenitor cells
Treating Vero cells with A. ureafaciens neuraminidase had no impact on ZIKV infection (Figure 1(f)), but reduced the infection of human influenza viruses that preferentially use α2,6-linked sialic acid as a receptor, by 3.3 and 2.8 log PFU/ml for H1N1 PR8 and A/ NWS/33 at 0.4 U/ml, respectively (Figure 1(g))

Summary

Introduction

Zika virus (ZIKV) is an arthropod-borne RNA virus in the genus Flavivirus with other vector-borne viruses significant to human health, such as dengue virus (DENV), yellow fever virus (YFV), West Nile virus (WNV), and Japanese encephalitis virus (JEV) [1]. Sporadic cases of ZIKV infections were reported over the half century before ZIKV emerged in major outbreaks in Yap Island in 2007 [3], French Polynesia in 2013 [4], and Brazil in 2015 [5]. These ZIKV outbreaks have been associated with Guillian-Barré syndrome and congenital microcephaly [6, 7]. The bona fide entry receptors for flaviviruses remain unknown, and many cell surface expressed molecules could contribute to infection These include C-type lectin DC-SIGN, L-SIGN, and phosphatidylserine receptors such as members of the T-cell Ig mucin (TIM)

Methods

Results

Conclusion