1. The use of green fluorescent protein-tagged virus-like particles as a tracer in the early phase of chikungunya infection.
- Author
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Tumkosit U, Maeda Y, Kishishita N, Siripanyaphinyo U, Omori H, Chetanachan P, Sittisaman P, Jityam C, Priengprom T, Mizushima H, Wongjaroen P, Mekada E, Tatsumi M, Takeda N, and Tanaka A
- Subjects
- Animals, Cells, Cultured, Chikungunya virus ultrastructure, Chlorocebus aethiops, Gene Expression, Genetic Vectors genetics, Models, Biological, Vero Cells, Viral Envelope Proteins chemistry, Viral Envelope Proteins genetics, Viral Envelope Proteins metabolism, Virus Internalization, Chikungunya Fever virology, Chikungunya virus physiology, Genes, Reporter, Green Fluorescent Proteins genetics, Virus Replication
- Abstract
To visually examine the early phase of chikungunya virus (CHIKV) infection in target cells, we constructed a virus-like particle (VLP) in which the envelope protein E1 is fused with green fluorescent protein (GFP). This chikungunya VLP-GFP (CHIK-VLP-EGFP), purified by density gradient fractionation, was observed as 60-70 nm-dia. particles and was detected as tiny puncta of fluorescence in the cells. CHIK-VLP-EGFP showed binding properties similar to those of the wild-type viruses. Most of the fluorescence signals that had bound on Vero cells disappeared within 30 min at 37 °C, but not in the presence of anti-CHIKV neutralizing serum or an endosomal acidification inhibitor (bafilomycin A1), suggesting that the loss of fluorescence signals is due to the disassembly of the viral envelope following the internalization of CHIK-VLP-EGFP. In addition to these results, the fluorescence signals disappeared in highly susceptible Vero and U251MG cells but not in poorly susceptible A549 cells. Thus, CHIK-VLP-EGFP is a useful tool to examine the effects of the CHIKV neutralizing antibodies and antiviral compounds that are effective in the entry phase of CHIKV., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2019
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