APOLIPOPROTEIN A1 AND B ENHANCE INTERNALIZATION AND INFECTION OF DENGUE VIRUS SEROTYPE 2 (DENV2) IN HUH-7 CELL LINES

Abstract

Intro: Previous studies have suggested an association between HDL, LDL cholesterol and dengue severity; however, the molecules involved in dengue virus (DENV) entry remain poorly understood. This study aims to determine the effects of apolipoproteins A1, B and E in DENV infectivity. Methods: Cytotoxicity of apolipoproteins on Huh-7 was assessed using MTS assays, incorporating varying concentrations of apolipoproteins (0.25- 2.00μg/ml). About 1.0 × 105 cells/well Huh-7 were seeded in 24-well plates and incubated at 37oC, 5% CO2 overnight. Subsequently, DENV2 at MOI of 1 with 2μg/ml apoA1, apoB, and apoE respectively were introduced and incubated for an hour. The cells were then washed once with PBS, replaced with 1% FBS supplemented fresh media and incubated for 72 hours. The supernatant was collected and subjected to qPCR for viral load measurement and will be compared to non-treated control (Huh-7 + DENV2). Findings: ApoA1, apoB and apoE concentrations of up to 2.00μg/ml show neutral effects on Huh-7 cell viability respectively. Treatment with apoA1, apoB and apoE increased DENV2 viral load compared to the non-treated control [apoA1=92.8%, apoB=76.9%, apoE=38.7%], where apoA1 and apoB demonstrated the highest and most significant increment of DENV2 internalization compared to apoE [p=0.004 and p=0.016 vs p= 0.897]. Conclusion: In conclusion, apolipoproteins (apoA1, apoB) enhances DENV internalization, indicating that both apoA1 and B are crucial for efficient DENV infection.