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Virus Genes

, Volume 49, Issue 2, pp 185–195 | Cite as

Recombinant dengue 2 virus NS3 protein conserves structural antigenic and immunological properties relevant for dengue vaccine design

  • Rosa Ramírez
  • Rosabel Falcón
  • Alienys Izquierdo
  • Angélica García
  • Mayling Alvarez
  • Ana Beatriz Pérez
  • Yudira Soto
  • Mayra Muné
  • Emiliana Mandarano da Silva
  • Oney Ortega
  • Ronaldo Mohana-Borges
  • María G. Guzmán
Article

Abstract

The NS3 protein is a multifunctional non-structural protein of flaviviruses implicated in the polyprotein processing. The predominance of cytotoxic T cell lymphocytes epitopes on the NS3 protein suggests a protective role of this protein in limiting virus replication. In this work, we studied the antigenicity and immunogenicity of a recombinant NS3 protein of the Dengue virus 2. The full-length NS3 gene was cloned and expressed as a His-tagged fusion protein in Escherichia coli. The pNS3 protein was purified by two chromatography steps. The recombinant NS3 protein was recognized by anti-protease NS3 polyclonal antibody and anti-DENV2 HMAF by Western Blot. This purified protein was able to stimulate the secretion of high levels of gamma interferon and low levels of interleukin-10 and tumor necrosis factor-α in mice splenocytes, suggesting a predominantly Th-1-type T cell response. Immunized BALB/c mice with the purified NS3 protein showed a strong induction of anti-NS3 IgG antibodies, essentially IgG2b, as determined by ELISA. Immunized mice sera with recombinant NS3 protein showed specific recognition of native dengue protein by Western blotting and immunofluorescence techniques. The successfully purified recombinant protein was able to preserv the structural and antigenic determinants of the native dengue protein. The antigenicity shown by the recombinant NS3 protein suggests its possible inclusion into future DENV vaccine preparations.

Keywords

Dengue virus NS3 Recombinant protein Immunogenicity Vaccine 

Notes

Acknowledgments

We would like to thank the International Society for Infectious Diseases (ISID) for providing financial support supports through an ISID Exchange Fellowship to Rosa Ramirez. We are grateful to technician Ramón Linares for his valuable help in the care and maintenance of the animals during the experiment. We also thank Melkis Alfonso, Osmel Fleitas and Tatiana Almaguer for the technical assistance. This study was also jointly supported by funds from the Cuban government, and CAPES, agency from de Brazilian government.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Rosa Ramírez
    • 1
  • Rosabel Falcón
    • 1
  • Alienys Izquierdo
    • 1
  • Angélica García
    • 1
  • Mayling Alvarez
    • 1
  • Ana Beatriz Pérez
    • 1
  • Yudira Soto
    • 1
  • Mayra Muné
    • 1
  • Emiliana Mandarano da Silva
    • 2
  • Oney Ortega
    • 1
  • Ronaldo Mohana-Borges
    • 2
  • María G. Guzmán
    • 1
  1. 1.PAHO/WHO Collaborating Centre for the Study of Dengue and its VectorInstitute of Tropical Medicine “Pedro Kourí”, IPKHavanaCuba
  2. 2.Laboratorio de Genômica Estrutural, Instituto de Biofísica Carlos Chagas FilhoUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil

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