Skip to main content

Advertisement

SpringerLink
Log in

Nucleocapsid-like particles of dengue-2 virus enhance the immune response against a recombinant protein of dengue-4 virus

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

In this study, we evaluate in mice a novel formulation containing nucleocapsid-like particles of dengue-2 virus (recNLP) co-immunized with a chimeric protein composed of the dengue-4 envelope domain III fused twice within the meningococcal P64k protein of Neisseria meningitidis (PD24). The animals receiving the PD24–recNLP mixture showed the highest levels of antiviral antibodies. Similar results were obtained for IFNγ secretion levels, indicating a functional Th1 cellular response. Consistently, the percentage of mice surviving after viral challenge was significantly higher for those immunized with the mixture than for those inoculated with PD24 protein alone. In addition, in vivo depletion experiments demonstrated the decisive role of CD4+ and CD8+ cells in the protection conferred by immunization with PD24–recNLP. In conclusion, this report demonstrates for the first time the adjuvant capacity of dengue-2 virus recNLP. Additionally, the evidence presented highlights the potential of these particles for enhancing the immune response against heterologous recombinant proteins.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Edelman R (2007) Dengue vaccines approach the finish line. Clin Infect Dis 45(Suppl 1):S56–S60

    Article  PubMed  Google Scholar 

  2. Grgacic EV, Anderson DA (2006) Virus-like particles, passport to immune recognition. Methods 40:60–65

    Article  CAS  PubMed  Google Scholar 

  3. Sugrue RJ, Fu J, Howe J, Chan YC (1997) Expression of the dengue virus structural proteins in Pichia pastoris leads to the generation of virus-like particles. J Gen Virol 78(Pt 8):1861–1866

    CAS  PubMed  Google Scholar 

  4. Pushko P, Tumpey TM, Bu F, Knell J, Robinson R, Smith G (2005) Influenza virus-like particles comprised of the HA, NA and M1 proteins of H9N2 influenza virus induce protective immune response in balb/c mice. Vaccine 23:5751–5759

    Article  CAS  PubMed  Google Scholar 

  5. Tacket CO, Sztein MB, Losonsky GA, Wasserman SS, Estes MK (2003) Humoral, mucosal, and cellular immune responses to oral Noralk virus-like particles in volunteers. Clin Immunol 108:241–247

    Article  CAS  PubMed  Google Scholar 

  6. Warfield K, Bosio C, Welcher B, Deal E, Mohamadzadeh M, Schmaljohn A, Aman MJ, Bavari S (2003) Ebola virus-like particles protect form lethal Ebola virus infection. Proc Natl Acad Sci USA 100:15889–15894

    Article  CAS  PubMed  Google Scholar 

  7. Saini M, Gratis S (2003) A Japanese encephalitis virus peptide present on Johnson grass mosaic virus-like particles induces virus-neutralizing antibodies and protects mice against lethal challenge. J Virol 77:3487–3494

    Article  CAS  PubMed  Google Scholar 

  8. Sadeyen J, Toume S, Shkreli M, Sizaret P, Coursaget P (2003) Insertion of a foreign sequence on capsid surface loops of human papillomavirus type 16 virus-like particles reduces their capacity to induce neutralizing antibodies and delineates a conformational neutralizing epitope. Virology 309:32–40

    Article  CAS  PubMed  Google Scholar 

  9. Kang S, Yao Q, Guo LCR (2009) Mucosal immunization with virus-like particles of Simian Immunodeficiency Virus conjugated with Cholera Toxin Subunit B. J Virol 77(18):9823–9830

    Article  Google Scholar 

  10. Alvarez-Lajonchere L, Gonzalez M, varez-Obregon JC, Guerra I, Vina A, costa-Rivero N, Musacchio A, Morales J, Duenas-Carrera S (2006) Hepatitis C virus (HCV) core protein enhances the immunogenicity of a co-delivered DNA vaccine encoding HCV structural antigens in mice. Biotechnol Appl Biochem 44:9–17

    Article  CAS  PubMed  Google Scholar 

  11. Storni T, Ruedl C, Schwarz K, Schwendener R, Renner W, Bachmann MF (2004) Nonmethylated CG motifs packaged into virus-like particles induce protective cytotoxic T cell responses in the absence of systemic side effects. J Immunol 172:1777–1785

    CAS  PubMed  Google Scholar 

  12. Lazo L, Hermida L, Zulueta A, Sanchez J, Lopez C, Silva R, Guillen G, Guzman MG (2007) A recombinant capsid protein from Dengue-2 induces protection in mice against homologous virus. Vaccine 25:1064–1070

    Article  CAS  PubMed  Google Scholar 

  13. Lopez C, Gil L, Lazo L, Menendez I, Marcos E, Sanchez J, Valdes I, Falcon V, de la Rosa MC, Marquez G, Guillen G, Hermida L (2009) In vitro assembly of nucleocapsid-like particles from purified recombinant capsid protein of dengue-2 virus. Arch Virol 154:695–698

    Article  CAS  PubMed  Google Scholar 

  14. Gil L, Lopez C, Lazo L, Valdes I, Marcos E, Alonso R, Gambe A, Martin J, Romero Y, Guzman MG, Guillen G, Hermida L (2009) Recombinant nucleocapsid-like particles from dengue-2 virus induce protective CD4+ and CD8+ cells against viral encephalitis in mice. Int Immunol 21(10):1175–1183

    Article  CAS  PubMed  Google Scholar 

  15. Zulueta A, Hermida L, Lazo L, Valdes I, Rodriguez R, Lopez C, Silva R, Rosario D, Martin J, Guzman MG, Guillen G (2003) The fusion site of envelope fragments from each serotype of Dengue virus in the P64k protein, influence some parameters of the resulting chimeric constructs. Biochem Biophys Res Commun 308:619–626

    Article  CAS  PubMed  Google Scholar 

  16. Hermida L, Rodriguez R, Lazo L, Silva R, Zulueta A, Chinea G, Lopez C, Guzman MG, Guillen G (2004) A dengue-2 Envelope fragment inserted within the structure of the P64k meningococcal protein carrier enables a functional immune response against the virus in mice. J Virol Methods 115:41–49

    Article  CAS  PubMed  Google Scholar 

  17. Bernardo L, Izquierdo A, Alvarez M, Rosario D, Prado I, Lopez C, Martinez R, Castro J, Santana E, Hermida L, Guillen G, Guzman MG (2008) Immunogenicity and protective efficacy of a recombinant fusion protein containing the domain III of the dengue 1 envelope protein in non-human primates. Antiviral Res 80:194–199

    Article  CAS  PubMed  Google Scholar 

  18. Gonzalez S, Alvarez A, Caballero E, Vina L, Guillen G, Silva R (2000) P64k meningococcal protein as immunological carrier for weak immunogens. Scand J Immunol 52:113–116

    Article  CAS  PubMed  Google Scholar 

  19. Perez A, Dickinson F, Cinza Z, Ruiz A, Serrano T, Sosa J, Gonzalez S, Gutierrez Y, Nazabal C, Gutierrez O, Guzman D, Diaz M, Delgado M, Caballero E, Sardinas G, Alvarez A, Martin A, Guillen G, Silva R (2001) Safety and preliminary immunogenicity of the recombinant outer membrane protein P64k of Neisseria meningitidis in human volunteers. Biotechnol Appl Biochem 34:121–125

    Article  CAS  PubMed  Google Scholar 

  20. Zulueta A, Martin J, Hermida L, Alvarez M, Valdes I, Prado I, Chinea G, Rosario D, Guillen G, Guzman MG (2006) Amino acid changes in the recombinant Dengue 3 envelope domain III determine its antigenicity and immunogenicity in mice. Virus Res 121:65–73

    Article  CAS  PubMed  Google Scholar 

  21. Roehrig JT (2003) Antigenic structure of flavivirus proteins. Adv Virus Res 59:141–175

    Article  CAS  PubMed  Google Scholar 

  22. Valdes I, Hermida L, Martin J, Menendez T, Gil L, Lazo L, Castro J, Niebla O, Lopez C, Bernardo L, Sanchez J, Romero Y, Martinez R, Guzman MG, Guillen G (2009) Immunological evaluation in nonhuman primates of formulations based on the chimeric protein P64k-domain III of dengue 2 and two components of Neisseria meningitidis. Vaccine 27:995–1001

    Article  CAS  PubMed  Google Scholar 

  23. Lazo L, Zulueta A, Hermida L, Blanco A, Sanchez J, Valdes I, Gil L, Lopez C, Romero Y, Guzman MG, Guillen G (2009) Dengue-4 envelope domain III fused twice within the meningococcal P64k protein carrier induces partial protection in mice. Biotechnol Appl Biochem 52:265–271

    Article  CAS  PubMed  Google Scholar 

  24. Kawano H, Rostapshov V, Rosen L, Lai CJ (1993) Genetic determinants of dengue type 4 virus neurovirulence for mice. J Virol 67:6567–6575

    CAS  PubMed  Google Scholar 

  25. Clarke DH, Casals J (1958) Techniques for hemagglutination and hemagglutination-inhibition with arthropod-borne viruses. Am J Trop Med Hyg 7:561–573

    CAS  PubMed  Google Scholar 

  26. Morens DM, Halstead SB, Repik PM, Putvatana R, Raybourne N (1985) Simplified plaque reduction neutralization assay for dengue viruses by semimicro methods in BHK-21 cells: comparison of the BHK suspension test with standard plaque reduction neutralization. J Clin Microbiol 22:250–254

    CAS  PubMed  Google Scholar 

  27. Kuhn RJ, Zhang W, Rossmann MG, Pletnev SV, Corver J, Lenches E, Jones CT, Mukhopadhyay S, Chipman PR, Strauss EG, Baker TS, Strauss JH (2002) Structure of dengue virus: implications for flavivirus organization, maturation, and fusion. Cell 108:717–725

    Article  CAS  PubMed  Google Scholar 

  28. Chen Y, Maguire T, Marks RM (1996) Demonstration of binding of dengue virus envelope protein to target cells. J Virol 70:8765–8772

    CAS  PubMed  Google Scholar 

  29. Mune M, Rodriguez R, Ramirez R, Soto Y, Sierra B, Rodriguez RR, Marquez G, Garcia J, Guillen G, Guzman MG (2003) Carboxy-terminally truncated Dengue 4 virus envelope glycoprotein expressed in Pichia pastoris induced neutralizing antibodies and resistance to Dengue 4 virus challenge in mice. Arch Virol 148:2267–2273

    Article  CAS  PubMed  Google Scholar 

  30. Simmons M, Murphy GS, Hayes CG (2001) Short report: Antibody responses of mice immunized with a tetravalent dengue recombinant protein subunit vaccine. Am J Trop Med Hyg 65:159–161

    CAS  PubMed  Google Scholar 

  31. Edelman R, Wasserman SS, Bodison SA, Putnak RJ, Eckels KH, Tang D, Kanesa-Thasan N, Vaughn DW, Innis BL, Sun W (2003) Phase I trial of 16 formulations of a tetravalent live-attenuated dengue vaccine. Am J Trop Med Hyg 69:48–60

    PubMed  Google Scholar 

  32. Vaughn DW (2000) Invited commentary: dengue lessons from Cuba. Am J Epidemiol 152:800–803

    Article  CAS  PubMed  Google Scholar 

  33. Putnak JR, de la Barrera R, Burgess T, Pardo J, Dessy F, Gheysen D, Lobet Y, Green S, Endy TP, Thomas SJ, Eckels KH, Innis BL, Sun W (2008) Comparative evaluation of three assays for measurement of dengue virus neutralizing antibodies. Am J Trop Med Hyg 79:115–122

    PubMed  Google Scholar 

  34. Chung YC, Ho MS, Wu JC, Chen WJ, Huang JH, Chou ST, Hu YC (2008) Immunization with virus-like particles of enterovirus 71 elicits potent immune responses and protects mice against lethal challenge. Vaccine 26:1855–1862

    CAS  PubMed  Google Scholar 

  35. Gagnon SJ, Zeng W, Kurane I, Ennis FA (1996) Identification of two epitopes on the dengue 4 virus capsid protein recognized by a serotype-specific and a panel of serotype-cross-reactive human CD4+ cytotoxic T-lymphocyte clones. J Virol 70:141–147

    CAS  PubMed  Google Scholar 

  36. Gagnon SJ, Ennis FA, Rothman AL (1999) Bystander target cell lysis and cytokine production by dengue virus-specific human CD4(+) cytotoxic T-lymphocyte clones. J Virol 73:3623–3629

    CAS  PubMed  Google Scholar 

  37. Mangada MM, Rothman AL (2005) Altered cytokine responses of dengue-specific CD4+ T cells to heterologous serotypes. J Immunol 175:2676–2683

    CAS  PubMed  Google Scholar 

  38. Mongkolsapaya J, Duangchinda T, Dejnirattisai W, Vasanawathana S, Avirutnan P, Jairungsri A, Khemnu N, Tangthawornchaikul N, Chotiyarnwong P, Sae-Jang K, Koch M, Jones Y, McMichael A, Xu X, Malasit P, Screaton G (2006) T cell responses in dengue hemorrhagic fever: are cross-reactive T cells suboptimal? J Immunol 176:3821–3829

    CAS  PubMed  Google Scholar 

  39. van der Most RG, Murali-Krishna K, Ahmed R (2003) Prolonged presence of effector-memory CD8 T cells in the central nervous system after dengue virus encephalitis. Int Immunol 15:119–125

    Article  PubMed  Google Scholar 

  40. van der Most RG, Murali-Krishna K, Ahmed R, Strauss JH (2000) Chimeric yellow fever/dengue virus as a candidate dengue vaccine: quantitation of the dengue virus-specific CD8 T-cell response. J Virol 74:8094–8101

    Article  Google Scholar 

  41. Iglesias E, Thompson R, Carrazana Y, Lobaina Y, Garcia D, Sanchez J, Garcia J, Cruz O, Brown E, Martin A, Muzio VL, Aguilar JC (2006) Coinoculation with hepatitis B surface and core antigen promotes a Th1 immune response to a multiepitopic protein of HIV-1. Immunol Cell Biol 84:174–183

    Article  CAS  PubMed  Google Scholar 

  42. Aguilar JC, Lobaina Y, Muzio V, Garcia D, Penton E, Iglesias E, Pichardo D, Urquiza D, Rodriguez D, Silva D, Petrovsky N, Guillen G (2004) Development of a nasal vaccine for chronic hepatitis B infection that uses the ability of hepatitis B core antigen to stimulate a strong Th1 response against hepatitis B surface antigen. Immunol Cell Biol 82:539–546

    Article  CAS  PubMed  Google Scholar 

  43. Chaung HC (2006) CpG oligodeoxynucleotides as DNA adjuvants in vertebrates and their applications in immunotherapy. Int Immunopharmacol 6:1586–1596

    Article  CAS  PubMed  Google Scholar 

  44. Riedl P, Stober D, Oehninger C, Melber K, Reimann J, Schirmbeck R (2002) Priming Th1 immunity to viral core particles is facilitated by trace amounts of RNA bound to its arginine-rich domain. J Immunol 168:4951–4959

    CAS  PubMed  Google Scholar 

  45. Donnelly J, Berry K, Ulmer JB (2003) Technical and regulatory hurdles for DNA vaccines. Int J Parasitol 33:457–467

    Article  CAS  PubMed  Google Scholar 

  46. Takeshita F, Leifer CA, Gursel I, Ishii KJ, Takeshita S, Gursel M, Klinman DM (2001) Cutting edge: role of Toll-like receptor 9 in CpG DNA-induced activation of human cells. J Immunol 167:3555–3558

    CAS  PubMed  Google Scholar 

  47. Lobigs M, Mullbacher A, Lee E (2004) Evidence that a mechanism for efficient flavivirus budding upregulates MHC class I. Immunol Cell Biol 82:184–188

    Article  CAS  PubMed  Google Scholar 

  48. Gil L, Lopez C, Blanco A, Lazo L, Martin J, Valdes I, Romero Y, Figueroa Y, Guillen G, Hermida L (2009) The cellular immune response plays an important role in protecting against dengue virus in the mouse encephalitis model. Viral Immunol 22:23–30

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Dr. Ricardo Silva, Dr. Eduardo Martínez-Montes and Dr. Alejandro Martín for their critical reading of the manuscript. This work was financed by the Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba.

Author information

Authors and Affiliations

  1. Vaccine Division, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box. 6162, 10600, Havana 6, Cuba

    Laura Lazo, Lázaro Gil, Carlos Lopez, Iris Valdes, Ernesto Marcos, Aracelys Blanco, Yaremis Romero, Viviana Falcon, Gerardo Guillén & Lisset Hermida

  2. PAHO/WHO Collaborating Center for Viral Diseases, Department of Virology, “Pedro Kourí” Tropical Medicine Institute (IPK), P.O. Box. 601, Havana, Cuba

    Mayling Álvarez & María G. Guzmán

Authors
  1. Laura Lazo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lázaro Gil
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carlos Lopez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Iris Valdes
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ernesto Marcos
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mayling Álvarez
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Aracelys Blanco
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yaremis Romero
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Viviana Falcon
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. María G. Guzmán
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Gerardo Guillén
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Lisset Hermida
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Laura Lazo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lazo, L., Gil, L., Lopez, C. et al. Nucleocapsid-like particles of dengue-2 virus enhance the immune response against a recombinant protein of dengue-4 virus. Arch Virol 155, 1587–1595 (2010). https://doi.org/10.1007/s00705-010-0734-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-010-0734-9

Keywords

Search

Navigation