Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journal

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Author(s)

  • Sanjay Sarkar
  • Lakshman Chelvarajan
  • Frank Cook
  • Sergey Artiushin
  • Shankar Mondal
  • Kelsi Anderson
  • John Eberth
  • Peter J. Timoney
  • Theodore S. Kalbfleisch
  • Ernest Bailey
  • Udeni B. R. Balasuriya

Detail(s)

Original languageEnglish
Pages (from-to)3366-3384
Journal / PublicationJournal of Virology
Volume90
Issue number7
Online published11 Mar 2016
Publication statusPublished - Apr 2016
Externally publishedYes

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.

Citation Format(s)

Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor. / Sarkar, Sanjay; Chelvarajan, Lakshman; Go, Yun Young; Cook, Frank; Artiushin, Sergey; Mondal, Shankar; Anderson, Kelsi; Eberth, John; Timoney, Peter J.; Kalbfleisch, Theodore S.; Bailey, Ernest; Balasuriya, Udeni B. R.

In: Journal of Virology, Vol. 90, No. 7, 04.2016, p. 3366-3384.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journal