An In Vitro Assay for Screening Interferons as Potent Inhibitors of Zika Virus Growth
Published: May 31, 2024
Abstract
Source: Contreras, D. et al., Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons. J. Vis. Exp. (2016)
The video demonstrates an in vitro assay for screening interferons with antiviral activity against the Zika virus, a human pathogen. Within the host cell, the Zika virus undergoes multiplication upon internalization, infecting neighboring cells and creating distinct zones or viral plaques. Type I interferon, acting as an antiviral agent, hinders virus multiplication, thereby preventing the formation of viral plaques in the host cell monolayer.
Protocol
1. Screening Cytokine Library against Zika Virus Infection Seed naive Vero cells at 1 x 105 cells per well using a 12-well plate. Once the cells are attached (6 hr post-seeding), add each cytokine at indicated concentrations in biological duplicates (2 ml volume/well). Include vehicle (PBS) alone control. At 12 hr post-treatment, perform Zika viral infection (MOI of 0.1 in 400 μl per well). Include negative control wells without infection (mock). At …
Representative Results
Figure 1: Interferons demonstrate potent anti-Zika viral activity. (A) Graph showing modulation of Zika viral growth by various cytokines compared to that of vehicle control. Mean values and standard deviation are shown in the bar graph. Interferons showed statistically significant inhibition of Zika virus replication (p value <0.05). (B and …
Divulgaciones
The authors have nothing to disclose.
Materials
| Vero Cell Line | ATCC | CCL-81 | |
| Zika viral strain PRVABC59 | Centers for Disease Control and Prevention (CDC) | ||
| IL-6 | Peprotech | 200-06 | |
| IL-1 alpha | Peprotech | 200-01A | |
| TNF-alpha | Peprotech | 300-01A | |
| Interferon alpha A | R & D Systems | 11100-1 | |
| Interferon beta | Peprotech | 300-02BC | |
| Interferon gamma | Peprotech | 300-02 | |
| Centrifuge 5415R | Eppendorf | 5415R | |
| Centrifuge 5810R | Eppendorf | 5810R | |
| Nikon Eclipse Ti Immunofluorescence Microscope with Nikon Intenselight C-HGFI | Nikon |
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An In Vitro Assay for Screening Interferons as Potent Inhibitors of Zika Virus Growth. J. Vis. Exp. (Pending Publication), e22274, doi: (2024).