Inhibition of Dengue Virus Entry and Multiplication into Monocytes Using RNA Interference

Overview

Journal PLoS Negl Trop Dis

Specialties Infectious Diseases
Tropical Medicine

Date 2011 Dec 6

PMID 22140591

Citations 24

Authors

Mohammed Abdelfatah Alhoot

Seok Mui Wang

Shamala Devi Sekaran

Affiliations

Soon will be listed here.

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.

Methodology/principal Findings: 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/significance: 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.

Citing Articles

Evaluation of Four Humanized NOD-Derived Mouse Models for Dengue Virus-2 Infection.

Gutierrez-Barbosa H, Medina-Moreno S, Perdomo-Celis F, Davis H, Chua J, Zapata J Pathogens. 2024; 13(8).

PMID: 39204240 PMC: 11357684. DOI: 10.3390/pathogens13080639.

A comprehensive overview on the crosstalk between microRNAs and viral pathogenesis and infection.

Bahojb Mahdavi S, Jebelli A, Aghbash P, Baradaran B, Amini M, Oroojalian F Med Res Rev. 2024; 45(2):349-425.

PMID: 39185567 PMC: 11796338. DOI: 10.1002/med.22073.

A Preliminary Investigation on the Antiviral Activities of the Philippine Marshmint (Mentha arvensis) Leaf Extracts against Dengue Virus Serotype 2 In Vitro.

Victoriano-Belvis A, Lao R, Morato M, Repotente E, Saclauso S, Inovejas S Kobe J Med Sci. 2022; 67(3):E98-E111.

PMID: 35367996 PMC: 9673882.

Elevated TNF-α Induces Thrombophagocytosis by Mononuclear Cells in ex vivo Whole-Blood Co-Culture with Dengue Virus.

Satria R, Jhan M, Chen C, Tseng P, Wang Y, Lin C J Inflamm Res. 2022; 15:1717-1728.

PMID: 35282270 PMC: 8906901. DOI: 10.2147/JIR.S356742.

Amaryllidaceae Alkaloid Cherylline Inhibits the Replication of Dengue and Zika Viruses.

Ka S, Merindol N, Sow A, Singh A, Landelouci K, Plourde M Antimicrob Agents Chemother. 2021; 65(9):e0039821.

PMID: 34152811 PMC: 8370201. DOI: 10.1128/AAC.00398-21.

References

Uprichard S . The therapeutic potential of RNA interference. FEBS Lett. 2005; 579(26):5996-6007. PMC: 7094730. DOI: 10.1016/j.febslet.2005.08.004. View

Ibanez C, Schrier R, Ghazal P, Wiley C, Nelson J . Human cytomegalovirus productively infects primary differentiated macrophages. J Virol. 1991; 65(12):6581-8. PMC: 250717. DOI: 10.1128/JVI.65.12.6581-6588.1991. View

Miller J, de Wet B, deWet B, Martinez-Pomares L, Radcliffe C, Dwek R . The mannose receptor mediates dengue virus infection of macrophages. PLoS Pathog. 2008; 4(2):e17. PMC: 2233670. DOI: 10.1371/journal.ppat.0040017. View

Ang F, Wong A, Ng M, Chu J . Small interference RNA profiling reveals the essential role of human membrane trafficking genes in mediating the infectious entry of dengue virus. Virol J. 2010; 7:24. PMC: 2825209. DOI: 10.1186/1743-422X-7-24. View

Orr S, Tobias P . LPS and LAM activation of the U373 astrocytoma cell line: differential requirement for CD14. J Endotoxin Res. 2000; 6(3):215-22. View

Subramanya S, Kim S, Abraham S, Yao J, Kumar M, Kumar P . Targeted delivery of small interfering RNA to human dendritic cells to suppress dengue virus infection and associated proinflammatory cytokine production. J Virol. 2009; 84(5):2490-501. PMC: 2820933. DOI: 10.1128/JVI.02105-08. View

Kwan W, Helt A, Maranon C, Barbaroux J, Hosmalin A, Harris E . Dendritic cell precursors are permissive to dengue virus and human immunodeficiency virus infection. J Virol. 2005; 79(12):7291-9. PMC: 1143643. DOI: 10.1128/JVI.79.12.7291-7299.2005. View

Chao Y, Huang C, Lee C, Chang S, King C, Kao C . Higher infection of dengue virus serotype 2 in human monocytes of patients with G6PD deficiency. PLoS One. 2008; 3(2):e1557. PMC: 2216061. DOI: 10.1371/journal.pone.0001557. View

Halstead S . Antibody, macrophages, dengue virus infection, shock, and hemorrhage: a pathogenetic cascade. Rev Infect Dis. 1989; 11 Suppl 4:S830-9. DOI: 10.1093/clinids/11.supplement_4.s830. View

10.

Clyde K, Kyle J, Harris E . Recent advances in deciphering viral and host determinants of dengue virus replication and pathogenesis. J Virol. 2006; 80(23):11418-31. PMC: 1642597. DOI: 10.1128/JVI.01257-06. View

11.

OSULLIVAN M, Killen H . The differentiation state of monocytic cells affects their susceptibility to infection and the effects of infection by dengue virus. J Gen Virol. 1994; 75 ( Pt 9):2387-92. DOI: 10.1099/0022-1317-75-9-2387. View

12.

Diamond M, Roberts T, Edgil D, Lu B, Ernst J, Harris E . Modulation of Dengue virus infection in human cells by alpha, beta, and gamma interferons. J Virol. 2000; 74(11):4957-66. PMC: 110847. DOI: 10.1128/jvi.74.11.4957-4966.2000. View

13.

Holland J . Evolving virus plagues. Proc Natl Acad Sci U S A. 1996; 93(2):545-6. PMC: 40086. DOI: 10.1073/pnas.93.2.545. View

14.

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A . Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7):RESEARCH0034. PMC: 126239. DOI: 10.1186/gb-2002-3-7-research0034. View

15.

Das S, Garver L, Ramirez J, Xi Z, Dimopoulos G . Protocol for dengue infections in mosquitoes (A. aegypti) and infection phenotype determination. J Vis Exp. 2008; (5):220. PMC: 2557096. DOI: 10.3791/220. View

16.

Padwad Y, Mishra K, Jain M, Chanda S, Karan D, Ganju L . RNA interference mediated silencing of Hsp60 gene in human monocytic myeloma cell line U937 revealed decreased dengue virus multiplication. Immunobiology. 2009; 214(6):422-9. DOI: 10.1016/j.imbio.2008.11.010. View

17.

Reyes-Del Valle J, Chavez-Salinas S, Medina F, Del Angel R . Heat shock protein 90 and heat shock protein 70 are components of dengue virus receptor complex in human cells. J Virol. 2005; 79(8):4557-67. PMC: 1069525. DOI: 10.1128/JVI.79.8.4557-4567.2005. View

18.

Hase T, Summers P, Eckels K . Flavivirus entry into cultured mosquito cells and human peripheral blood monocytes. Arch Virol. 1989; 104(1-2):129-43. DOI: 10.1007/BF01313814. View

19.

Tuschl T . RNA interference and small interfering RNAs. Chembiochem. 2002; 2(4):239-45. DOI: 10.1002/1439-7633(20010401)2:4<239::AID-CBIC239>3.0.CO;2-R. View

20.

Peng T, Wang J, Chen W, Zhang J, Gao N, Chen Z . Entry of dengue virus serotype 2 into ECV304 cells depends on clathrin-dependent endocytosis, but not on caveolae-dependent endocytosis. Can J Microbiol. 2009; 55(2):139-45. DOI: 10.1139/w08-107. View