CD24 Expression Dampens the Basal Antiviral State in Human Neuroblastoma Cells and Enhances Permissivity to Zika Virus Infection
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Lines, Viruses, and Infections
2.2. Plaque Assays
2.3. Immunostaining and Fluorescence Microscopy
2.4. Quantification of Virus Infection, Cell Viability, and CD24
2.5. Exogenous Treatment with IFN-I or IFN-III and Quantification of IFN-I Levels
2.6. Reverse Transcription and Real-Time PCR
2.7. Nuclear Extraction
2.8. Western Blotting and Densitometry
2.9. Supernatant Transfer
2.10. Global Transcriptomics
2.11. Figures, Statistics, and Images
3. Results
3.1. CD24 Expression Increases ZIKV Spread and Infectivity in a Neuroblastoma Cell Line
3.2. STAT1 Is Constitutively Phosphorylated and Localized to the Nucleus in CD24-Low Cells
3.3. CD24 Expression Decreases the Transcriptional Response of CD24-Low Cells to IFN-I Stimulation and Increases Their Permissivity to RNA Virus Infection
3.4. IRF-1 and NFκB Levels Are Elevated in CD24-Low Cells Compared to CD24-High Cells
3.5. CD24-Low and -High Cells Do Not Differ in Basal IFN-I Production or Soluble Antiviral Mediators
3.6. Ectopic Expression of CD24 Alters the Global Transcriptomic Profile of Intracellular Signaling Pathways
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Forward Primer | Reverse Primer |
---|---|---|
GAPDH | 5′-TTAAAAGCAGCCCTGGTGAC-3′ | 5′-CTCTGCTCCTGTTCGAC-3′ |
Ns1 | 5′-GCCATCACAATACCAGAGAG-3′ | 5′-GGCCTTATCTCCATTCCATAC-3′ |
IFN-β | 5′-CAGCTCTTTCCATGAGCTACAA-3′ | 5′-CAGTATTCAAGCCTCCCATTCA-3′ |
IFITM3 | 5′-ACCATTCTGCTCATCGTCATC-3′ | 5′-GAAGTTGGAGTACGTGGGATAC-3′ |
OAS2 | 5′-AGAAGCTGGGTTGGTTTATC-3′ | 5′-GACGTCACAGATGGTGTTC-3′ |
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Kedarinath, K.; Fox, C.R.; Crowgey, E.; Mazar, J.; Phelan, P.; Westmoreland, T.J.; Alexander, K.A.; Parks, G.D. CD24 Expression Dampens the Basal Antiviral State in Human Neuroblastoma Cells and Enhances Permissivity to Zika Virus Infection. Viruses 2022, 14, 1735. https://doi.org/10.3390/v14081735
Kedarinath K, Fox CR, Crowgey E, Mazar J, Phelan P, Westmoreland TJ, Alexander KA, Parks GD. CD24 Expression Dampens the Basal Antiviral State in Human Neuroblastoma Cells and Enhances Permissivity to Zika Virus Infection. Viruses. 2022; 14(8):1735. https://doi.org/10.3390/v14081735
Chicago/Turabian StyleKedarinath, Kritika, Candace R. Fox, Erin Crowgey, Joseph Mazar, Peter Phelan, Tamarah J. Westmoreland, Kenneth A. Alexander, and Griffith D. Parks. 2022. "CD24 Expression Dampens the Basal Antiviral State in Human Neuroblastoma Cells and Enhances Permissivity to Zika Virus Infection" Viruses 14, no. 8: 1735. https://doi.org/10.3390/v14081735