An Assay to Evaluate the Effect of Test Compounds on Viral Entry and Fusion in Host Cells

Published: February 29, 2024

Abstract

Source: Tai, C., et al. Early Viral Entry Assays for the Identification and Evaluation of Antiviral Compounds. J. Vis. Exp. (2015).

This video demonstrates an assay to evaluate antiviral agents that target specific steps of the viral entry into host cells and fusion with the endosome. The cells are incubated at a low temperature and then infected with HCV. The low temperature permits viral binding to host cells but inhibits internalization via endocytosis. The test antiviral compound is added, and the cells are incubated at physiological temperature. This shift to a higher temperature facilitates viral entry and allows assessment of the impact of the test compound on viral envelope fusion with endosomes.

Protocol

Note: Ensure that all procedures involving cell culture and virus infection are conducted in certified biosafety hoods that are appropriate for the biosafety level of the samples being handled. For the purpose of describing the protocols, Gaussia luciferase reporter-tagged HCV is used as a model virus. In the context of the representative results, the compounds chebulagic acid (CHLA) and punicalagin (PUG) are used as candidate antivirals that target viral glycoprotein interactions with cell surface glyco…

Representative Results

Table 1: Host cell type for viral infection. The cell type used for each viral infection system described in the representative results is indicated. Virus Cell Type HCMV HEL HCV Huh-7.5 DENV-2 Vero <…

Disclosures

The authors have nothing to disclose.

Materials

DMEM GIBCO 11995-040
FBS GIBCO 26140-079
Penicillin-Streptomycin GIBCO 15070-063
Amphotericin B GIBCO 15290-018
DMSO Sigma D5879
In vitro toxicology assay kit, XTT-based Sigma TOX2
PBS pH 7.4 GIBCO 10010023
Microplate reader Bio-Tek Instrument, Inc. ELx800
Microcentrifuge Thermo Scientific 75002420
BioLux Gaussia luciferase assay kit New England Biolabs E3300L
Luminometer Promega GloMax-20/20
Sodium citrate, dihydrate Sigma 71402
Potassium chloride Sigma P5405

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Cite This Article
An Assay to Evaluate the Effect of Test Compounds on Viral Entry and Fusion in Host Cells. J. Vis. Exp. (Pending Publication), e22000, doi: (2024).

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