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Archives of Virology

, Volume 154, Issue 6, pp 919–928 | Cite as

A clinical isolate of dengue virus and its proteins induce apoptosis in HMEC-1 cells: a possible implication in pathogenesis

  • Melina Vásquez Ochoa
  • Julio García Cordero
  • Benito Gutiérrez Castañeda
  • Leopoldo Santos Argumedo
  • Nicolas Villegas Sepúlveda
  • Leticia Cedillo Barrón
Original Article

Abstract

Cumulative studies have demonstrated that dengue virus infection results in the induction of apoptosis of certain cells in vitro. Moreover, apoptosis of microvascular endothelial cells in the brain and in the intestinal serosa has been demonstrated postmortem in dengue virus (DENV)-infected patients. In this work, human microvascular endothelial cells (HMEC-1) infected with a DENV-2 clinical isolate, or HMEC-1 cells transfected with its protease sequence (NS3pro) or its complex (NS2BNS3pro) were able to trigger apoptosis after 24 h of infection or transfection. The infected or transfected HMEC-1 cells displayed the distinctive apoptotic hallmarks, which include cytoplasmic shrinkage and plasma membrane blebbing. In addition, the transfected HMEC-1 cells showed biochemical changes such as exposure of phosphatidylserine on the outer leaflet of the plasma membrane, TUNEL positivity, caspase 3 activation and cleaved PARP, a central regulator of apoptosis. These findings suggest the role of such proteins from the clinical isolate in the induction of apoptosis.

Keywords

Viral Protein Dengue Virus Microvascular Endothelial Cell DENV Infection Human Microvascular Endothelial Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We are thankful to Fernando Medina for his assistance in C6/36 culture, and Julio García Cordero (185263) and Melina Vasquez Ochoa (158086), who received predoctoral fellowships from CONACYT. This work was supported by CONACYT (grant no. 40199) and by DENFRAME Consortium INCO 517711.

References

  1. 1.
    Andrews BS, Theofilopoulos AN, Peters CJ, Loskutoff DJ, Brandt WE, Dixon FJ (1978) Replication of dengue and junin viruses in cultured rabbit and human endothelial cells. Infect Immun 20:776–781PubMedGoogle Scholar
  2. 2.
    Avirutnan P, Malasit P, Seliger B, Bhakdi S, Husmann M (1998) Dengue virus infection of human endothelial cells leads to chemokine production, complement activation, and apoptosis. J Immunol 161:6338–6346PubMedGoogle Scholar
  3. 3.
    Basu A, Chaturvedi UC (2008) Vascular endothelium: the battlefield of dengue viruses. FEMS Immunol Med Microbiol 53:287–299PubMedCrossRefGoogle Scholar
  4. 4.
    Bosch I, Xhaja K, Estevez L, Raines G, Melichar H, Warke RV, Fournier MV, Ennis FA, Rothman AL (2002) Increased production of interleukin-8 in primary human monocytes and in human epithelial and endothelial cell lines after dengue virus challenge. J Virol 76:5588–5597PubMedCrossRefGoogle Scholar
  5. 5.
    Bunyaratvej A, Butthep P, Yoksan S, Bhamarapravati N (1997) Dengue viruses induce cell proliferation and morphological changes of endothelial cells. Southeast Asian J Trop Med Public Health 28(Suppl 3):32–37PubMedGoogle Scholar
  6. 6.
    Cardier JE, Marino E, Romano E, Taylor P, Liprandi F, Bosch N, Rothman AL (2005) Proinflammatory factors present in sera from patients with acute dengue infection induce activation and apoptosis of human microvascular endothelial cells: possible role of TNF-alpha in endothelial cell damage in dengue. Cytokine 30:359–365PubMedCrossRefGoogle Scholar
  7. 7.
    Catteau A, Kalinina O, Wagner MC, Deubel V, Courageot MP, Despres P (2003) Dengue virus M protein contains a proapoptotic sequence referred to as ApoptoM. J Gen Virol 84:2781–2793PubMedCrossRefGoogle Scholar
  8. 8.
    Catteau A, Roue G, Yuste VJ, Susin SA, Despres P (2003) Expression of dengue ApoptoM sequence results in disruption of mitochondrial potential and caspase activation. Biochimie 85:789–793PubMedCrossRefGoogle Scholar
  9. 9.
    Chen SO, Chang TJ, Stone G, Chen CH, Liu JJ (2006) Programmed cell death induced by Japanese encephalitis virus YL vaccine strain or its recombinant envelope protein in varied cultured cells. Intervirology 49:346–351PubMedCrossRefGoogle Scholar
  10. 10.
    Clarke P, Tyler KL (2009) Apoptosis in animal models of virus-induced disease. Nat Rev Microbiol 7:144–155PubMedCrossRefGoogle Scholar
  11. 11.
    Couvelard A, Marianneau P, Bedel C, Drouet MT, Vachon F, Henin D, Deubel V (1999) Report of a fatal case of dengue infection with hepatitis: demonstration of dengue antigens in hepatocytes and liver apoptosis. Hum Pathol 30:1106–1110PubMedCrossRefGoogle Scholar
  12. 12.
    Darzynkiewicz Z, Traganos F (1998) Measurement of apoptosis. Adv Biochem Eng Biotechnol 62:33–73PubMedGoogle Scholar
  13. 13.
    Despres P, Frenkiel MP, Ceccaldi PE, Duarte Dos Santos C, Deubel V (1998) Apoptosis in the mouse central nervous system in response to infection with mouse-neurovirulent dengue viruses. J Virol 72:823–829PubMedGoogle Scholar
  14. 14.
    Duarte dos Santos CN, Frenkiel MP, Courageot MP, Rocha CF, Vazeille-Falcoz MC, Wien MW, Rey FA, Deubel V, Despres P (2000) Determinants in the envelope E protein and viral RNA helicase NS3 that influence the induction of apoptosis in response to infection with dengue type 1 virus. Virology 274:292–308PubMedCrossRefGoogle Scholar
  15. 15.
    Honda M, Kaneko S, Shimazaki T, Matsushita E, Kobayashi K, Ping LH, Zhang HC, Lemon SM (2000) Hepatitis C virus core protein induces apoptosis and impairs cell-cycle regulation in stably transformed Chinese hamster ovary cells. Hepatology 31:1351–1359PubMedCrossRefGoogle Scholar
  16. 16.
    Huang YH, Lei HY, Liu HS, Lin YS, Liu CC, Yeh TM (2000) Dengue virus infects human endothelial cells and induces IL-6 and IL-8 production. Am J Trop Med Hyg 63:71–75PubMedGoogle Scholar
  17. 17.
    Jan JT, Chen BH, Ma SH, Liu CI, Tsai HP, Wu HC, Jiang SY, Yang KD, Shaio MF (2000) Potential dengue virus-triggered apoptotic pathway in human neuroblastoma cells: arachidonic acid, superoxide anion, and NF-kappaB are sequentially involved. J Virol 74:8680–8691PubMedCrossRefGoogle Scholar
  18. 18.
    Jessie K, Fong MY, Devi S, Lam SK, Wong KT (2004) Localization of dengue virus in naturally infected human tissues, by immunohistochemistry and in situ hybridization. J Infect Dis 189:1411–1418PubMedCrossRefGoogle Scholar
  19. 19.
    Krishnamurti C, Peat RA, Cutting MA, Rothwell SW (2002) Platelet adhesion to dengue-2 virus-infected endothelial cells. Am J Trop Med Hyg 66:435–441PubMedGoogle Scholar
  20. 20.
    Kurane I (2007) Dengue hemorrhagic fever with special emphasis on immunopathogenesis. Comp Immunol Microbiol Infect Dis 30:329–340PubMedCrossRefGoogle Scholar
  21. 21.
    Labat-Moleur F, Guillermet C, Lorimier P, Robert C, Lantuejoul S, Brambilla E, Negoescu A (1998) TUNEL apoptotic cell detection in tissue sections: critical evaluation and improvement. J Histochem Cytochem 46:327–334PubMedGoogle Scholar
  22. 22.
    Lam M, Stallcup M, Distelhorst CW (1997) Expression of a defective mouse mammary tumor virus envelope glycoprotein precursor which binds stably to GRP78 within the lumen of the endoplasmic reticulum is associated with decreased glucocorticoid-induced apoptosis in mouse lymphoma cells. Cell Death Differ 4:283–288PubMedCrossRefGoogle Scholar
  23. 23.
    Lanciotti RS, Calisher CH, Gubler DJ, Chang GJ, Vorndam AV (1992) Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 30:545–551PubMedGoogle Scholar
  24. 24.
    Lei HY, Yeh TM, Liu HS, Lin YS, Chen SH, Liu CC (2001) Immunopathogenesis of dengue virus infection. J Biomed Sci 8:377–388PubMedCrossRefGoogle Scholar
  25. 25.
    Leitmeyer KC, Vaughn DW, Watts DM, Salas R, Villalobos I, de C, Ramos C, Rico-Hesse R (1999) Dengue virus structural differences that correlate with pathogenesis. J Virol 73:4738–4747PubMedGoogle Scholar
  26. 26.
    Liew KJ, Chow VT (2004) Differential display RT-PCR analysis of ECV304 endothelial-like cells infected with dengue virus type 2 reveals messenger RNA expression profiles of multiple human genes involved in known and novel roles. J Med Virol 72:597–609PubMedCrossRefGoogle Scholar
  27. 27.
    Limon-Flores AY, Perez-Tapia M, Estrada-Garcia I, Vaughan G, Escobar-Gutierrez A, Calderon-Amador J, Herrera-Rodriguez SE, Brizuela-Garcia A, Heras-Chavarria M, Flores-Langarica A, Cedillo-Barron L, Flores-Romo L (2005) Dengue virus inoculation to human skin explants: an effective approach to assess in situ the early infection and the effects on cutaneous dendritic cells. Int J Exp Pathol 86:323–334PubMedCrossRefGoogle Scholar
  28. 28.
    Limonta D, Capo V, Torres G, Perez AB, Guzman MG (2007) Apoptosis in tissues from fatal dengue shock syndrome. J Clin Virol 40:50–54PubMedCrossRefGoogle Scholar
  29. 29.
    Lindenbach BD, Rice CM (2003) Molecular biology of flaviviruses. Adv Virus Res 59:23–61PubMedCrossRefGoogle Scholar
  30. 30.
    Marianneau P, Cardona A, Edelman L, Deubel V, Despres P (1997) Dengue virus replication in human hepatoma cells activates NF-kappaB which in turn induces apoptotic cell death. J Virol 71:3244–3249PubMedGoogle Scholar
  31. 31.
    Marusawa H, Hijikata M, Chiba T, Shimotohno K (1999) Hepatitis C virus core protein inhibits Fas- and tumor necrosis factor alpha-mediated apoptosis via NF-kappaB activation. J Virol 73:4713–4720PubMedGoogle Scholar
  32. 32.
    Moorman JP, Prayther D, McVay D, Hahn YS, Hahn CS (2003) The C-terminal region of hepatitis C core protein is required for Fas-ligand independent apoptosis in Jurkat cells by facilitating Fas oligomerization. Virology 312:320–329PubMedCrossRefGoogle Scholar
  33. 33.
    Negoescu A, Lorimier P, Labat-Moleur F, Drouet C, Robert C, Guillermet C, Brambilla C, Brambilla E (1996) In situ apoptotic cell labeling by the TUNEL method: improvement and evaluation on cell preparations. J Histochem Cytochem 44:959–968PubMedGoogle Scholar
  34. 34.
    Nicholson DW, Ali A, Thornberry NA, Vaillancourt JP, Ding CK, Gallant M, Gareau Y, Griffin PR, Labelle M, Lazebnik YA et al (1995) Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 376:37–43PubMedCrossRefGoogle Scholar
  35. 35.
    Prikhod’ko GG, Prikhod’ko EA, Pletnev AG, Cohen JI (2002) Langat flavivirus protease NS3 binds caspase-8 and induces apoptosis. J Virol 76:5701–5710PubMedCrossRefGoogle Scholar
  36. 36.
    Prikhod’ko EA, Prikhod’ko GG, Siegel RM, Thompson P, Major ME, Cohen JI (2004) The NS3 protein of hepatitis C virus induces caspase-8-mediated apoptosis independent of its protease or helicase activities. Virology 329:53–67PubMedCrossRefGoogle Scholar
  37. 37.
    Ramanathan MP, Chambers JA, Pankhong P, Chattergoon M, Attatippaholkun W, Dang K, Shah N, Weiner DB (2006) Host cell killing by the West Nile Virus NS2B-NS3 proteolytic complex: NS3 alone is sufficient to recruit caspase-8-based apoptotic pathway. Virology 345:56–72PubMedCrossRefGoogle Scholar
  38. 38.
    Rico-Hesse R, Harrison LM, Salas RA, Tovar D, Nisalak A, Ramos C, Boshell J, de Mesa MT, Nogueira RM, da Rosa AT (1997) Origins of dengue type 2 viruses associated with increased pathogenicity in the Americas. Virology 230:244–251PubMedCrossRefGoogle Scholar
  39. 39.
    Roehrig JT (2003) Antigenic structure of flavivirus proteins. Adv Virus Res 59:141–175PubMedCrossRefGoogle Scholar
  40. 40.
    Sacco R, Tsutsumi T, Suzuki R, Otsuka M, Aizaki H, Sakamoto S, Matsuda M, Seki N, Matsuura Y, Miyamura T, Suzuki T (2003) Antiapoptotic regulation by hepatitis C virus core protein through up-regulation of inhibitor of caspase-activated DNase. Virology 317:24–35PubMedCrossRefGoogle Scholar
  41. 41.
    Shafee N, AbuBakar S (2003) Dengue virus type 2 NS3 protease and NS2B-NS3 protease precursor induce apoptosis. J Gen Virol 84:2191–2195PubMedCrossRefGoogle Scholar
  42. 42.
    Su HL, Lin YL, Yu HP, Tsao CH, Chen LK, Liu YT, Liao CL (2001) The effect of human bcl-2 and bcl-X genes on dengue virus-induced apoptosis in cultured cells. Virology 282:141–153PubMedCrossRefGoogle Scholar
  43. 43.
    Thongtan T, Panyim S, Smith DR (2004) Apoptosis in dengue virus infected liver cell lines HepG2 and Hep3B. J Med Virol 72:436–444PubMedCrossRefGoogle Scholar
  44. 44.
    Van Cruchten S, Van Den Broeck W (2002) Morphological and biochemical aspects of apoptosis, oncosis and necrosis. Anat Histol Embryol 31:214–223PubMedCrossRefGoogle Scholar
  45. 45.
    Warke RV, Xhaja K, Martin KJ, Fournier MF, Shaw SK, Brizuela N, de Bosch N, Lapointe D, Ennis FA, Rothman AL, Bosch I (2003) Dengue virus induces novel changes in gene expression of human umbilical vein endothelial cells. J Virol 77:11822–11832PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Melina Vásquez Ochoa
    • 1
  • Julio García Cordero
    • 1
  • Benito Gutiérrez Castañeda
    • 2
  • Leopoldo Santos Argumedo
    • 1
  • Nicolas Villegas Sepúlveda
    • 1
  • Leticia Cedillo Barrón
    • 1
  1. 1.Departamento de Biomedicina MolecularCentro de Investigación y de Estudios AvanzadosMexico DFMexico
  2. 2.Laboratorio de Inmunología UMF de la FES IztacalaUniversidad Autónoma de MéxicoTlanepantlaMexico

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