Vol. 47, No. 2, 2004
Issue release date: March–April 2004
Intervirology 2004;47:78–86
Original Paper
Add to my selection

Internalization and Propagation of the Dengue Virus in Human Hepatoma (HepG2) Cells

Thepparit C. · Phoolcharoen W. · Suksanpaisan L. · Smith D.R.
Molecular Pathology Laboratory, Institute of Molecular Biology and Genetics, Mahidol University, Nakorn Pathom, Thailand
email Corresponding Author

 goto top of outline Key Words

  • Flavivirus
  • Glycosaminoglycans
  • Infection
  • Liver

 goto top of outline Abstract

Objectives: This study sought to undertake a comparative analysis of the internalization and propagation of all four dengue serotypes in a single cell line of human liver origin, HepG2. Methods: Virus production after infection was determined by the plaque assay technique. Internalization profiles were determined by incubating virus and cells on ice and then raising the temperature for various times. The contribution of extracellular matrix components to internalization was determined by pretreatment of cells with either trypsin or heparinase III. Results: HepG2 cells were able to support the propagation of all four serotypes with mature viruses being produced by 12 h for dengue serotype 4 and by 17–18 h for the remaining serotypes. Virus internalization showed a plateau for serotypes 1, 2 and 4 entry while serotype 3 showed a constant increase in internalization for up to 5 h. Pretreatment of HepG2 cells with heparinase III or trypsin both resulted in a reduction in viral production, with the smallest effect being noted for dengue serotype 3. Conclusion: These results suggest that the interaction between the dengue virus and liver cells is a complex one that requires both protein and nonprotein elements, and has a significant serotype/strain element.

Copyright © 2004 S. Karger AG, Basel

 goto top of outline References
  1. Hales S, de Wet N, Maindonald J, Woodward A: Potential effect of population and climate changes on global distribution of dengue fever: An empirical model. Lancet 2002;360:830–834.
  2. Burke DS, Nisalak A, Johnson DE, Scott RM: A prospective study of dengue infections in Bangkok. Am J Trop Med Hyg 1988;381:172–180.
  3. Halstead SB: Antibody, macrophages, dengue virus infection, shock, and hemorrhage: A pathogenetic cascade. Rev Infect Dis 1989(suppl 4):S830–839.
  4. Guzman MG, Kouri G, Bravo J, Valdes L, Vazquez S, Halstead SB: Effect of age on outcome of secondary dengue 2 infections. Int J Infect Dis 2002;6:118–124.
  5. Pinheiro FP, Corber SJ: Global situation of dengue and dengue haemorrhagic fever, and its emergence in the Americas. World Health Stat Q 1997;50:161–169.
  6. Halstead SB, O’Rourke EJ: Dengue viruses and mononuclear phagocytes. I. Infection enhancement by non-neutralizing antibody. J Exp Med 1977;146:201–217.
  7. Halstead SB, Nimmannitya S, Yamarat C, Russell PK: Hemorrhagic fever in Thailand; recent knowledge regarding etiology. Jpn J Med Sci Biol 1967;20:96–103.
  8. Russell PK, Yuill TM, Nisalak A, Udomsakdi S, Gould DJ, Winter PE: An insular outbreak of dengue hemorrhagic fever. II. Virologic and serologic studies. Am J Trop Med Hyg 1968;17:600–608.
  9. Sangkawibha N, Rojanasuphot S, Ahandrik S, Viriyapongse S, Jatanasen S, Salitul V, Phanthumachinda B, Halstead SB: Risk factors in dengue shock syndrome: A prospective epidemiologic study in Rayong, Thailand. I. The 1980 outbreak. Am J Epidemiol 1984;120:653–669.
  10. Mohan B, Patwari AK, Anand VK: Hepatic dysfunction in childhood dengue infection. J Trop Pediatr 2000;46:40–43.
  11. Couvelard A, Marianneau P, Bedel C, Drouet MT, Vachon F, Henin D, Deubel V: Report of a fatal case of dengue infection with hepatitis: Demonstration of dengue antigens in hepatocytes and liver apoptosis. Hum Pathol 1999;30:1106–1110.
  12. Nguyen TL, Nguyen TH, Tieu NT: The impact of dengue haemorrhagic fever on liver function. Res Virol 1997;148:273–277.
  13. Rosen L, Khin MM, U T: Recovery of virus from the liver of children with fatal dengue: Reflections on the pathogenesis of the disease and its possible analogy with that of yellow fever. Res Virol 1989;140:351–360.
  14. Kuo CH, Tai DI, Chang-Chien CS, Lan CK, Chiou SS, Liaw YF: Liver biochemical tests and dengue fever. Am J Trop Med Hyg 1992;47:265–270.
  15. Edelman R, Nimmannitya S, Colman RW, Talamo RC, Top FH Jr: Evaluation of the plasma kinin system in dengue hemorrhagic fever. J Lab Clin Med 1975;86:410–421.
  16. Huerre MR, Lan NT, Marianneau P, Hue NB, Khun H, Hung NT, Khen NT, Drouet MT, Huong VT, Ha DQ, Buisson Y, Deubel V: Liver histopathology and biological correlates in five cases of fatal dengue fever in Vietnamese children. Virchows Arch 2001;438:107–115.
  17. Marianneau P, Flamand M, Deubel V, Despres P: Apoptotic cell death in response to dengue virus infection: The pathogenesis of dengue haemorrhagic fever revisited. Clin Diagn Virol 1998;10:113–119.
  18. Bhamarapravati N, Tuchinda P, Boonyapaknavik V: Pathology of Thailand haemorrhagic fever: A study of 100 autopsy cases. Ann Trop Med Parasitol 1967;61:500–510.
  19. Bielefeldt-Ohmann H, Meyer M, Fitzpatrick DR, Mackenzie JS: Dengue virus binding to human leukocyte cell lines: Receptor usage differs between cell types and virus strains. Virus Res 2001;73:81–89.
  20. Hung SL, Lee PL, Chen HW, Chen LK, Kao CL, King CC: Analysis of the steps involved in dengue virus entry into host cells. Virology 1999;257:156–167.
  21. Long D, Cohen GH, Muggeridge MI, Eisenberg RJ: Cysteine mutants of herpes simplex virus type 1 glycoprotein D exhibit temperature-sensitive properties in structure and function. J Virol 1990;64:5542–5552.
  22. Hase T, Summers PL, Eckels KH: Flavivirus entry into cultured mosquito cells and human peripheral blood monocytes. Arch Virol 1989;104:129–143.
  23. Hilgard P, Stockert R: Heparan sulfate proteoglycans initiate dengue virus infection of hepatocytes. Hepatology 2000;32:1069–1077.
  24. Germi R, Crance JM, Garin D, Guimet J, Lortat-Jacob H, Ruigrok RW, Zarski JP, Drouet E: Heparan sulfate-mediated binding of infectious dengue virus type 2 and yellow fever virus. Virology 2002;292:162–168.
  25. Lin YL, Lei HY, Lin YS, Yeh TM, Chen SH, Liu HS: Heparin inhibits dengue-2 virus infection of five human liver cell lines. Antiviral Res 2002;56:93–96.
  26. Lin YL, Liu CC, Lei HY, Yeh TM, Lin YS, Chen RM, Liu HS: Infection of five human liver cell lines by dengue-2 virus. J Med Virol 2000;60:425–431.
  27. Marianneau P, Cardona A, Edelman L, Deubel V, Despres P: Dengue virus replication in human hepatoma cells activates NF-kappaB which in turn induces apoptotic cell death. J Virol 1997;71:3244–3249.
  28. Diamond MS, Edgil D, Roberts TG, Lu B, Harris E: Infection of human cells by dengue virus is modulated by different cell types and viral strains. J Virol 2000;74:7814–7823.
  29. Barth OM, Schatzmayr HG: Brazilian dengue virus type 1 replication in mosquito cell cultures. Mem Inst Oswaldo Cruz 1992;87:1–7.
  30. Se-Thoe SY, Ling AE, Ng MM: Alteration of virus entry mode: A neutralisation mechanism for dengue-2 virus. J Med Virol 2000;62:364–376.
  31. Lim HY, Ng ML: A different mode of entry by dengue-2 neutralisation escape mutant virus. Arch Virol 1999;144:989–995.
  32. Daughaday CC, Brandt WE, McCown JM, Russell PK: Evidence for two mechanisms of dengue virus infection of adherent human monocytes: Trypsin-sensitive virus receptors and trypsin-resistant immune complex receptors. Infect Immun 1981;32:469–473.
  33. Chen Y, Maguire T, Hileman RE, Fromm JR, Esko JD, Linhardt RJ, Marks RM: Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate. Nat Med 1997;3:866–871.
  34. Chen YC, Wang SY, King CC: Bacterial lipopolysaccharide inhibits dengue virus infection of primary human monocytes/macrophages by blockade of virus entry via a CD14-dependent mechanism. J Virol 1999;73:2650–2657.
  35. Moreno-Altamirano MM, Sanchez-Garcia FJ, Munoz ML: Non-Fc receptor-mediated infection of human macrophages by dengue virus serotype 2. J Gen Virol 2002;83:1123–1130.
  36. Martinez-Barragan JJ, del Angel RM: Identification of a putative coreceptor on Vero cells that participates in dengue 4 virus infection. J Virol 2001;75:7818–7827.
  37. Ramos-Castaneda J, Imbert JL, Barron BL, Ramos C: A 65-kDa trypsin-sensible membrane cell protein as a possible receptor for dengue virus in cultured neuroblastoma cells. J Neurovirol 1997;3:435–440.

 goto top of outline Author Contacts

Duncan R. Smith
Molecular Pathology Laboratory, Institute of Molecular Biology and Genetics
Mahidol University, Salaya Campus, 25/25 Phuttamonthol Sai 4
Salaya, Nakorn Pathom, 73170 (Thailand)
Tel. +66 2 441 9003, Fax +66 2 441 9906, E-Mail duncan_r_smith@hotmail.com

 goto top of outline Article Information

Received: July 16, 2003
Accepted after revision: September 5, 2003
Number of Print Pages : 9
Number of Figures : 5, Number of Tables : 0, Number of References : 37

 goto top of outline Publication Details

Intervirology (International Journal of Basic and Medical Virology)
Founded 1973 by J.L. Melnick; continued by F. Rapp (1986–1990); M.J. Buchmeier and C.R. Howard (1991–1993)

Vol. 47, No. 2, Year 2004 (Cover Date: March-April 2004)

Journal Editor: Rüdiger W. Braun, Stuttgart
ISSN: 0300–5526 (print), 1423–0100 (Online)

For additional information: http://www.karger.com/journals/int

Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.