title: Differential humoral and cellular immunity induced by vaccination using plasmid DNA and protein recombinant expressing the NS3 protein of dengue virus type 3.
abstract: BACKGROUND
  The dengue non-structural 3 (NS3) is a multifunctional protein, containing a serine-protease domain, located at the N-terminal portion, and helicase, NTPase and RTPase domains present in the C-terminal region. This protein is considered the main target for CD4+ and CD8+ T cell responses during dengue infection, which may be involved in protection. However, few studies have been undertaken evaluating the use of this protein as a protective antigen against dengue, as well as other flavivirus. In the present work we evaluated the potential of the NS3 (protease domain) as a protective antigen by comparing the administration of a recombinant protein versus a DNA vaccine in the mouse model.
  
  
  RESULTS
  BALB/c mice were immunized with the recombinant protein NS3-DEN3 via intraperitoneal and with plasmid pcDNA3/NS3-DEN3 intramuscularly and the immune response was evaluated. The activity of T lymphocytes was analyzed by the MTT assay, and cells of mice immunized with the recombinant protein showed no activity when stimulated with the homologous protein. However, cells from mice immunized with DNA, responded to stimulation with the recombinant protein. When the expression (RT-PCR) and cytokine production (ELISA) was evaluated in the splenocytes, different behavior depending on the type of immunization was observed, splenocytes of mice immunized with the recombinant protein expressed cytokines such as IL-4, IL-10 and produced high concentrations of IL-1, IL-6 and TNFα. Splenocytes from mice immunized with DNA expressed IL-2 and IFNγ and did not produce IL-6. In addition, immunization with the recombinant protein induced the production of antibodies that are detected up to a dilution 1:3200 by ELISA and Western blot assays, however, the serum of mice immunized with DNA presented no detectable antibody titers.
  
  
  CONCLUSIONS
  The results obtained in this study show that administration of pcDNA3/NS3-DEN3 induces a favorable response in the activation of T lymphocytes with low production of specific antibodies against NS3-DEN3.
authors.affiliation: Doctorado en Ciencias Biomédicas, Universidad Veracruzana, Xalapa, Veracruz, Mexico.
authors.forename: M L
authors.initials: ML
authors.lastname: Hurtado-Melgoza
authors.affiliation: LADISER Inmunología y Biología Molecular, Facultad de Ciencias Químicas, Universidad Veracruzana, Orizaba, Veracruz, Mexico. angramos@uv.mx.|Centro de Investigaciones Biomédicas, Universidad Veracruzana, Xalapa, Veracruz, Mexico. angramos@uv.mx.
authors.forename: A
authors.initials: A
authors.lastname: Ramos-Ligonio
authors.affiliation: LADISER Inmunología y Biología Molecular, Facultad de Ciencias Químicas, Universidad Veracruzana, Orizaba, Veracruz, Mexico.
authors.forename: L M
authors.initials: LM
authors.lastname: Álvarez-Rodríguez
authors.affiliation: Facultad de Medicina, Universidad Veracruzana, Xalapa, Veracruz, Mexico.
authors.forename: T
authors.initials: T
authors.lastname: Meza-Menchaca
authors.affiliation: LADISER Inmunología y Biología Molecular, Facultad de Ciencias Químicas, Universidad Veracruzana, Orizaba, Veracruz, Mexico.|Centro de Investigaciones Biomédicas, Universidad Veracruzana, Xalapa, Veracruz, Mexico.
authors.forename: A
authors.initials: A
authors.lastname: López-Monteon
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003715:Dengue / Q000517:prevention & control / Q000821:virology; D003716:Dengue Virus / Q000276:immunology* / Q000472:pathogenicity; D006801:Humans; D007111:Immunity, Cellular; D056724:Immunity, Humoral; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage / Q000276:immunology; D020365:RNA Helicases / Q000008:administration & dosage / Q000276:immunology; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology; D012697:Serine Endopeptidases / Q000008:administration & dosage / Q000276:immunology; D014611:Vaccination; D017361:Viral Nonstructural Proteins / Q000008:administration & dosage / Q000276:immunology*; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000956:Antigens, Viral; D020365:RNA Helicases; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
keywords: DNA vaccine; Dengue virus; NS3 protein; Recombinant protein