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West‐Nile virus replicon particles infect 293T cells expressing Dendritic Cell‐Specific Intercellular Adhesion Molecule‐3‐Grabbing Non‐Integrin Related.
Author(s) -
Leonard Nathan,
Crowson Nicole,
Madigan Amanda,
Schmitz Madilyn,
Alcala Elana,
Martinez Osvaldo
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.658.1
Subject(s) - replicon , virology , biology , tropism , virus , tissue tropism , viral entry , hek 293 cells , flavivirus , viral replication , transfection , integrin , cell culture , microbiology and biotechnology , cell , gene , genome , genetics
West‐Nile virus (WNV) is an arthropod‐borne virus that is frequently transmitted to humans through the bite of an infected mosquito. Less than one percent of cases result in a serious neurological illness such as meningitis or encephalitis. To investigate WNV tropism we have established several 293T stable cell lines expressing dendritic cell‐specific intercellular adhesion molecule‐3‐grabbing non‐integrin related (DC‐SIGNR) or DC‐SIGN. Previous studies have suggested that WNV infection efficiency is enhanced by expression of DC‐SIGNR, but not DC‐SIGN on target cells. We demonstrate high levels of DC‐SIGNR expression in selected 293T cells. Replication incompetent virus replicon particles (VRPs) are used to safely and conveniently study viral entry. Successful VRP entry into target cells transfers a mutated incomplete viral genome containing the GFP gene which is expressed from the infected cells. Infections assays using WNV (NY99) VRPs as well as a variant of WNV (NY99) which contains the beta‐lactamase enzyme, show significant infection in DC‐SIGNR expressing, but not control cells that do not express DC‐SIGNR. The establishment of this entry assay system will allow us to safely study virus tropism. Support or Funding Information Student Research Grants, Winona State University Winona, MN