Profiling of Microrna Expression within the Cells of Peripheral Blood Mononuclear after an Infection with Serotype-2 of Dengue Virus: Preliminary Study

Abstract

The role of microRiboNucleic Acids (miRNA), a small-non coding RNA has been associated with immune regulation in various viral infectionincluding dengue infection. The microRNA will bind a specific protein target in order to encourage an explosive expression of various cytokines, known as cytokines storm in Dengue infection.The objective of this study aimed to determine and evaluate themicroRNAs profile expression withinperipheral blood mononuclear cells having been infected with one of the dengue virus serotype.To obtained the PBMCs from a healthy donor, Ficoll density gradient centrifugation was used to isolate the PBMCs and then followed infecting it with a DENV-2 clinical isolate. Prior to PBMCs isolation, the virus has been propagated and having titration to get an optimal virus titer. We conducted the infection at the multiplication of infections 4 PFU/106 cells.MiRCURYLNATMExiqon was utilized on purpose to extract the RNA. Quantitative Real-Time PCR was applied in order for the miRNAs relative expression to be measured. The preliminary result reveals that miR-150, miR-146a, hsa-let-7e expression were increased 1.74 folds, 2 folds, and 1.49 foldsrespectively at 12 hours post-infection on PBMCs upon DENV-2 infection.The expression of microRNAswas discovered to behigher inPBMCsat the time of infection withDENV-2.ThemiRNAs expression in the uninfected PMBCs was lower than that of the miRNA. This high expression of miRNAsin dengue infection may proceedto dengue infection pathogenesis.

Host and viral non‐coding RNAs in dengue pathogenesis

“…miR-146a, miR-150, let-7e was reported to be induced upon DENV infection suggesting their role in dengue pathogenesis. 43 miR-146a was found to facilitate DENV replication in the human monocytes and macrophage cell line. 35 However, another study identified no effect of miR-146a overexpression of DENV replication in human lung cell-line, A549, possibly due to a cell-type specific effect of miRNAs.…”

“…35 However, another study identified no effect of miR-146a overexpression of DENV replication in human lung cell-line, A549, possibly due to a cell-type specific effect of miRNAs. 44 Similarly, induction of hsa-let-7c, hsa-miR-146a differs in DENV infection ex-vivo in PBMCs [41][42][43] compared to DENV2 infection in human macrophage cell lines. 35,39 DENV infection progression is systematic and dynamic under the infection.…”

“…The role of miRNAs in DENV pathogenesis is explored mostly through in vitro studies. miR‐146a, miR‐150, let‐7e was reported to be induced upon DENV infection suggesting their role in dengue pathogenesis 43 . miR‐146a was found to facilitate DENV replication in the human monocytes and macrophage cell line 35 .…”

Differential expression of microRNA, miR-150 and enhancer of zeste homolog 2 (EZH2) in peripheral blood cells as early prognostic markers of severe forms of dengue

“…Similarly, miR-30e also expresses differentially in dengue infected cultured cells [24]. miR-150 expression is upregulated in DENV infected cultured cells within few hours of the infection [25]. Expression of miR-150 is also found to be significantly upregulated in SD patients compared to DF patients [26].…”

Differential expression of selected microRNA and putative target genes in peripheral blood cells as early markers of severe forms of dengue

“…Similarly, miR-30e-3p also express differentially in dengue infected cultured cells [24]. miR-150 expression is upregulated in DENV infected cultured cells within few hours of the infection [25]. Expression of miR150 is also found to be significantly upregulated in DHF patients compared to DF patients [26].…”