Equine arteritis virus uses equine CXCL16 as an entry receptor

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

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

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.

Original languageEnglish
Pages (from-to)3366-3384
Number of pages19
JournalJournal of Virology
Volume90
Issue number7
DOIs
StatePublished - 2016

Bibliographical note

Publisher Copyright:
© 2016, American Society for Microbiology.

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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