THE REPLICATION EFFICIENCY OF DENGUE VIRUS SEROTYPE 1 ISOLATED FROM PATIENTS WITH DENGUE FEVER IN HUMAN HEPATOCYTE CELL LINES.

Abstract

Background: The Efficiency of viral replication in cells depends on the capability of supporting virus replication by the cells. We characterized the effectiveness of Dengue Virus Serotype 1 (DENV-1) replication in various cell lines and various multiplicity of infection (MOI) starting from 2 FFU/cell up to 0,3125 FFU/cell.

Materials and method: We used HepG2 and Huh-7 human hepatocyte cell lines and in addition, we also used non-human kidney cells (Vero cells). DENV-1 strain IDS 11/2010 was isolated from DF patients and previously propagated in Huh7 and Vero cells as DENV-1-adapted Huh-7 and DENV-1-adapted Vero cells, respectively. Huh7 cells, Hep G2 cells, and Vero cells were infected with DENV-1 at various MOI and incubated for 48 hours at 37⁰C with 5% CO₂. DENV-infected cells were determined by indirect immuno-peroxidase staining using 3,3'-Diaminobenzidine (DAB). DENV-1 infected cells as foci were counted under inverted light microscopy and were used to determine the virus titer.

Results: The virus was adapted to Huh-7 and Vero cells, with results showing that Vero cells exhibited the highest replication efficiency, evidenced by significant viral titers. Among human hepatocyte cell lines, DENV-1 demonstrated greater replication in Huh-7 cells than in HepG2 cells. Notably, no foci formation was observed in HepG2 cells after 48 hours of infection.

Conclusion: These findings underscore the suitability of Vero and Huh-7 cells as optimal environments for DENV-1 replication, offering valuable insights for enhancing laboratory diagnostics and advancing antiviral strategies and vaccine development against DENV-1.