Inhibition of dengue virus entry and multiplication into monocytes using RNA interference

• Published in: PLoS neglected tropical diseases Vol. 5; no. 11; p. e1410
• Main Authors: Alhoot, Mohammed Abdelfatah, Wang, Seok Mui, Sekaran, Shamala Devi
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.11.2011; Public Library of Science (PLoS)
• Subjects: Analysis; Biology; Biomedical research; Blood banks; Blood Donors; Bone marrow; Care and treatment; Cells, Cultured; Clathrin; Clathrin Heavy Chains - antagonists & inhibitors; Clathrin Heavy Chains - genetics; Dengue; Dengue fever; Dengue Virus - pathogenicity; Development and progression; Disease; Dynamin II - antagonists & inhibitors; Dynamin II - genetics; Endocytosis; Fever; Flow cytometry; Gene expression; Gene Silencing; Health aspects; Humans; Infections; Lipopolysaccharide Receptors - genetics; Lipopolysaccharide Receptors - metabolism; Medicine; Monocytes - physiology; Monocytes - virology; Multiplication & division; Physiological aspects; Prevention; Proteins; Public health; RNA Interference; Studies; Tropical diseases; Viral Load; Virus Internalization; Malaysia; California; Dynamin II; Antigens, CD14; Humans; Cells, Cultured; Gene Silencing; Dengue Virus; Viral Load; Virus Internalization; Blood Donors; Endocytosis; Monocytes; RNA Interference; Clathrin Heavy Chains
• ISSN: 1935-2735; 1935-2727; 1935-2735
• DOI: 10.1371/journal.pntd.0001410

Dengue infection ranks as one of the most significant viral diseases of the globe. Currently, there is no specific vaccine or antiviral therapy for prevention or treatment. Monocytes/macrophages are the principal target cells for dengue virus and are responsible for disseminating the virus after its transmission. Dengue virus enters target cells via receptor-mediated endocytosis after the viral envelope protein E attaches to the cell surface receptor. This study aimed to investigate the effect of silencing the CD-14 associated molecule and clathrin-mediated endocytosis using siRNA on dengue virus entry into monocytes. Gene expression analysis showed a significant down-regulation of the target genes (82.7%, 84.9 and 76.3% for CD-14 associated molecule, CLTC and DNM2 respectively) in transfected monocytes. The effect of silencing of target genes on dengue virus entry into monocytes was investigated by infecting silenced and non-silenced monocytes with DENV-2. Results showed a significant reduction of infected cells (85.2%), intracellular viral RNA load (73.0%), and extracellular viral RNA load (63.0%) in silenced monocytes as compared to non-silenced monocytes. Silencing the cell surface receptor and clathrin mediated endocytosis using RNA interference resulted in inhibition of the dengue virus entry and subsequently multiplication of the virus in the monocytes. This might serve as a novel promising therapeutic target to attenuate dengue infection and thus reduce transmission as well as progression to severe dengue hemorrhagic fever.