Apolipoproteins and Its Association with Dengue Virus Serotype 2 (DENV-2) Infectivity in Human Hepatoma Cell Line (Huh7)
DOI:
https://doi.org/10.48048/tis.2024.7674Keywords:
Dengue, Apolipoprotein, apoA1, apoB, apoEAbstract
Introduction: Previous studies have shown that lipid components are involved in the internalization of Dengue virus (DENV), however, this mechanism has been hypothesized mostly through clinical studies correlating between dengue severity and lipid levels. This study aimed to determine DENV-2 internalization in the presence of apolipoprotein A1 (apoA1), B (apoB) or E (apoE) in human hepatoma cell line (Huh7). Materials and methods: Tetrazolium reduction assays were used to assess the cytotoxicity of apolipoproteins on Huh7. DENV-2 at MOI of 1 with 2 µg/mL apoA1, apoB and apoE, respectively were introduced into the confluent Huh7 cells and incubated for an hour (internalization) and 72 h (replication) at 37 °C, 5 % CO2. The replication experiments were repeated after SR-B1 siRNA treatment. Samples collected were subjected to qPCR for viral load determination. Differences between groups were assessed by ANOVA and independent T-Test. Results and discussion: Treatment with apoA1 and apoB demonstrated significant increment in DENV-2 internalization and replication compared to apoE (internalization: p = 0.022 and p = 0.323 vs p = 0.878, respectively; replication: p = 0.004 and p = 0.016 vs p = 0.897, respectively). A significant reduction of DENV2 infectivity was seen in apoA1 treated cells with SR-B1 down-regulation/silencing (35.2 % reduction, p = 0.013). Conclusions: apoA1 and apoB, enhanced initial attachment of DENV-2 to cell surface, suggesting their role in DENV internalization and infectivity.
HIGHLIGHTS
- This research determined the effects of apolipoproteins on dengue virus serotype 2 (DENV-2) internalization in vitro using the human hepatoma cell line (Huh7)
- apoA1 and apoB enhance the initial attachment of DENV-2 to the cell surface
- Knockdown of SR-B1 receptors reduces apoA1-treated DENV-2 internalization, suggesting the role of SR-B1 as a route of entry for DENV-2
- Knockdown of SR-B1 receptors had no effect on apoB-treated DENV-2 internalization, suggesting it uses another entry pathway
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