Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor

Authors

Sanjay Sarkar, University of KentuckyFollow
Lakshman Chelvarajan, University of KentuckyFollow
Yun Young Go, University of KentuckyFollow
Frank Cook, University of KentuckyFollow
Sergey Artiushin, University of KentuckyFollow
Shankar Mondal, University of Kentucky
Kelsi Anderson, University of KentuckyFollow
John E. Eberth, University of KentuckyFollow
Peter J. Timoney, University of KentuckyFollow
Theodore S. Kalbfleisch, University of Louisville
Ernest F. Bailey, University of KentuckyFollow
Udeni B. R. Balasuriya, University of KentuckyFollow

Abstract

Previous studies in our laboratory have identified equine CXCL16 (EqCXCL16) to be a candidate molecule and possible cell entry receptor for equine arteritis virus (EAV). In horses, the CXCL16 gene is located on equine chromosome 11 (ECA11) and encodes a glycosylated, type I transmembrane protein with 247 amino acids. Stable transfection of HEK-293T cells with plasmid DNA carrying EqCXCL16 (HEK-EqCXCL16 cells) increased the proportion of the cell population permissive to EAV infection from < 3% to almost 100%. The increase in permissiveness was blocked either by transfection of HEK-EqCXCL16 cells with small interfering RNAs (siRNAs) directed against EqCXCL16 or by pretreatment with guinea pig polyclonal antibody against EqCXCL16 protein (Gp anti-EqCXCL16 pAb). Furthermore, using a virus overlay protein-binding assay (VOPBA) in combination with far-Western blotting, gradient-purified EAV particles were shown to bind directly to the EqCXCL16 protein in vitro. The binding of biotinylated virulent EAV strain Bucyrus at 4°C was significantly higher in HEK-EqCXCL16 cells than nontransfected HEK-293T cells. Finally, the results demonstrated that EAV preferentially infects subpopulations of horse CD14⁺ monocytes expressing EqCXCL16 and that infection of these cells is significantly reduced by pretreatment with Gp anti-EqCXCL16 pAb. The collective data from this study provide confirmatory evidence that the transmembrane form of EqCXCL16 likely plays a major role in EAV host cell entry processes, possibly acting as a primary receptor molecule for this virus.

Document Type

Article

Publication Date

4-2016

Notes/Citation Information

Published in Journal of Virology, v. 90, no. 7, p. 3366-3384.

Copyright © 2016, American Society for Microbiology. All Rights Reserved.

The copyright holders have granted the permission for posting the article here.

Digital Object Identifier (DOI)

https://doi.org/10.1128/JVI.02455-15

Funding Information

USDA | National Institute of Food and Agriculture (NIFA) provided funding to Udeni B. R Balasuriya under grant number 2013-68004-20360.

Repository Citation

Sarkar, Sanjay; Chelvarajan, Lakshman; Go, Yun Young; Cook, Frank; Artiushin, Sergey; Mondal, Shankar; Anderson, Kelsi; Eberth, John E.; Timoney, Peter J.; Kalbfleisch, Theodore S.; Bailey, Ernest F.; and Balasuriya, Udeni B. R., "Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor" (2016). Veterinary Science Faculty Publications. 34.
https://uknowledge.uky.edu/gluck_facpub/34