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Published on: June 5, 2020
Internalization and propagation of the dengue virus in human hepatoma (HepG2) cells
Chutima Thepparit1, Waranyoo Phoolcharoen, Lukkana Suksanpaisan
1Molecular Pathology Laboratory, Institute of Molecular Biology and Genetics, Mahidol University, Nakorn Pathom, Thailand.
Objectives:
This study sought to undertake a comparative analysis of the internalization and propagation of all four dengue serotypes in a single cell line of human liver origin, HepG2.
Methods:
Virus production after infection was determined by the plaque assay technique. Internalization profiles were determined by incubating virus and cells on ice and then raising the temperature for various times. The contribution of extracellular matrix components to internalization was determined by pretreatment of cells with either trypsin or heparinase III.
Results:
HepG2 cells were able to support the propagation of all four serotypes with mature viruses being produced by 12 h for dengue serotype 4 and by 17-18 h for the remaining serotypes. Virus internalization showed a plateau for serotypes 1, 2 and 4 entry while serotype 3 showed a constant increase in internalization for up to 5 h. Pretreatment of HepG2 cells with heparinase III or trypsin both resulted in a reduction in viral production, with the smallest effect being noted for dengue serotype 3.
Conclusion:
These results suggest that the interaction between the dengue virus and liver cells is a complex one that requires both protein and nonprotein elements, and has a significant serotype/strain element.
Insights
This study shows that all four dengue serotypes infect and replicate in HepG2 liver cells, with differences in internalization and replication rates. Heparinase III and trypsin treatments reveal complex interactions involving protein and non-protein elements.
Area of Science:
- Virology
- Hepatology
- Cell Biology
Background:
- Dengue virus poses a significant global health threat.
- Understanding dengue virus-host interactions is crucial for therapeutic development.
- Human liver cells, such as HepG2, are potential targets for dengue virus infection.
Purpose of the Study:
- To comparatively analyze the internalization and propagation of all four dengue serotypes in HepG2 cells.
- To investigate the role of extracellular matrix components in dengue virus entry into liver cells.
Main Methods:
- Dengue virus serotypes 1-4 were used to infect HepG2 cells.
- Virus production was quantified using the plaque assay technique.
- Internalization kinetics were assessed by temperature shift experiments.
- The role of extracellular matrix was evaluated using trypsin and heparinase III pre-treatments.
Main Results:
- HepG2 cells supported the propagation of all four dengue serotypes.
- Dengue serotype 4 showed faster virus production (12h) compared to others (17-18h).
- Dengue serotype 3 exhibited distinct internalization kinetics, with a continuous increase for 5 hours, unlike other serotypes.
- Heparinase III and trypsin treatments reduced viral production, indicating involvement of protein and non-protein matrix components.
Conclusions:
- HepG2 cells are a suitable model for studying dengue virus infection.
- Dengue virus-host cell interactions in liver cells are complex and serotype-dependent.
- Both protein and non-protein elements of the extracellular matrix play a role in dengue virus internalization and replication.
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