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A Murine Model of Dengue Virus-induced Acute Viral Encephalitis-like Disease
Published on: April 28, 2019
Protegrin-1 inhibits dengue NS2B-NS3 serine protease and viral replication in MK2 cells
Hussin A Rothan1, Ammar Y Abdulrahman, Pottayil G Sasikumer
1Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. molec.genetics@gmail.com
Abstract:
Dengue diseases have an economic as well as social burden worldwide. In this study, the antiviral activity of protegrin-1 (PG-1, RGGRLCYCRRRFCVCVGR) peptide towards dengue NS2B-NS3pro and viral replication in Rhesus monkey kidney (MK2) cells was investigated. The peptide PG-1 was synthesized by solid-phase peptide synthesis, and disulphide bonds formation followed by peptide purification was confirmed by LC-MS and RPHPLC. Dengue NS2B-NS3pro was produced as a single-chain recombinant protein in E. coli. The NS2B-NS3pro assay was carried out by measuring the florescence emission of catalyzed substrate. Real-time PCR was used to evaluate the inhibition potential of PG-1 towards dengue serotype-2 (DENV-2) replication in MK2 cells. The results showed that PG-1 inhibited dengue NS2B-NS3pro at IC(50) of 11.7 μM. The graded concentrations of PG-1 at nontoxic range were able to reduce viral replication significantly (P < 0.001) at 24, 48, and 72 hrs after viral infection. However, the percentage of inhibition was significantly (P < 0.01) higher at 24 hrs compared to 48 and 72 hrs. These data show promising therapeutic potential of PG-1 against dengue infection, hence it warrants further analysis and improvement of the peptide features as a prospective starting point for consideration in designing attractive dengue virus inhibitors.
Insights
The antiviral peptide protegrin-1 (PG-1) effectively inhibits dengue virus NS2B-NS3 protease activity and significantly reduces viral replication in cell cultures. Further research into PG-1 shows potential for developing new dengue virus inhibitors.
Area of Science:
- Virology
- Biochemistry
- Drug Discovery
Background:
- Dengue virus poses a significant global health and economic burden.
- Existing treatments for dengue virus are limited, necessitating novel therapeutic strategies.
- Antiviral peptides are emerging as promising candidates for infectious disease treatment.
Purpose of the Study:
- To investigate the antiviral activity of protegrin-1 (PG-1) against dengue virus.
- To evaluate PG-1's inhibitory effects on the dengue virus NS2B-NS3 protease.
- To assess PG-1's efficacy in reducing dengue virus replication in cell culture.
Main Methods:
- Solid-phase peptide synthesis was used to create PG-1.
- Recombinant dengue virus NS2B-NS3 protease was expressed in E. coli.
- Enzyme inhibition assays and real-time PCR were employed to measure antiviral activity and viral replication.
Main Results:
- PG-1 demonstrated significant inhibition of dengue virus NS2B-NS3 protease with an IC50 of 11.7 μM.
- Non-toxic concentrations of PG-1 substantially reduced dengue virus serotype-2 replication in Rhesus monkey kidney cells.
- The highest inhibition percentage of viral replication was observed at 24 hours post-infection.
Conclusions:
- Protegrin-1 exhibits potent antiviral activity against dengue virus.
- PG-1 shows promise as a therapeutic agent for dengue infection.
- Further optimization of PG-1 could lead to the development of novel dengue virus inhibitors.
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