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Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
Published on: November 1, 2018
Gefitinib and pyrrolidine dithiocarbamate decrease viral replication and cytokine production in dengue virus infected
Anyelo Duran1, Nereida Valero2, Jesús Mosquera3
1Instituto de Investigaciones Clínicas "Dr. Américo Negrette", Facultad de Medicina, Universidad del Zulia, Maracaibo, Venezuela; Cátedra de Bioquímica General, Escuela de Bioanálisis, Facultad de Medicina, Universidad del Zulia, Maracaibo, Venezuela; Sociedad Venezolana de Microbiología, Venezuela.
Gefitinib (GBTC) and pyrrolidine dithiocarbamate (PDTC) reduced dengue virus type 2 (DENV2) replication and pro-inflammatory cytokine production in human monocytes. These findings suggest potential therapeutic benefits for dengue treatment by targeting EGFR/NOD2 and NF-kB pathways.
Area of Science:
- Virology
- Immunology
- Pharmacology
Background:
- Epidermal growth factor receptor (EGFR) and nucleotide-binding and oligomerization-domain containing 2 (NOD2) are implicated in cancer and microbial recognition, respectively.
- Both EGFR and NOD2 activate NF-kB signaling, leading to inflammatory gene expression.
- Monocytes are crucial for early dengue virus (DENV) replication and cytokine release.
Purpose of the Study:
- To investigate the effects of Gefitinib (GBTC), an EGFR inhibitor, and pyrrolidine dithiocarbamate (PDTC), an NF-kB inhibitor, on DENV serotype 2 (DENV2) replication.
- To assess the impact of GBTC and PDTC on cytokine production in DENV2-infected human monocyte cultures.
Main Methods:
- Human monocyte cultures were infected with DENV2.
- Inhibition of EGFR/NOD2 was achieved using GBTC, and NF-kB inhibition using PDTC.
- Cytokine production was quantified using ELISA, and viral replication was measured by plaque-forming unit (PFU) assays.
Main Results:
- DENV2 infection led to increased viral replication and the production of antiviral cytokines (IFN-α/β, TNF-α, IL-12, IL-18).
- Inhibition of EGFR/NOD2 with GBTC or NF-kB with PDTC significantly decreased DENV2 replication.
- Both GBTC and PDTC treatments reduced the production of pro-inflammatory cytokines in infected monocytes.
Conclusions:
- GBTC and PDTC demonstrate inhibitory effects on DENV2 replication and associated cytokine production.
- These findings highlight a potential therapeutic role for targeting EGFR/NOD2 and NF-kB pathways in managing dengue infection.
- The study suggests that EGFR/NOD2 receptors and NF-kB signaling are involved in the pathogenesis of dengue virus infection.

