title: Inhibition of dengue virus entry and multiplication into monocytes using RNA interference.
abstract: BACKGROUND
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Mohammed Abdelfatah
authors.initials: MA
authors.lastname: Alhoot
authors.forename: Seok Mui
authors.initials: SM
authors.lastname: Wang
authors.forename: Shamala Devi
authors.initials: SD
authors.lastname: Sekaran
mesh_terms: D001782:Blood Donors; D002478:Cells, Cultured; D033922:Clathrin Heavy Chains / Q000037:antagonists & inhibitors* / Q000235:genetics; D003716:Dengue Virus / Q000472:pathogenicity*; D034285:Dynamin II / Q000037:antagonists & inhibitors* / Q000235:genetics; D004705:Endocytosis; D020868:Gene Silencing; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000235:genetics / Q000378:metabolism*; D009000:Monocytes / Q000502:physiology / Q000821:virology*; D034622:RNA Interference; D019562:Viral Load; D053586:Virus Internalization
chemical_list: C101272:CLTCL1 protein, human; D018950:Lipopolysaccharide Receptors; D033922:Clathrin Heavy Chains; D034285:Dynamin II