Differential humoral and cellular immunity induced by vaccination using plasmid DNA and protein recombinant expressing the NS3 protein of dengue virus type 3

Abstract

BackgroundThe 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.ResultsBALB/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.ConclusionThe 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.

Long-Term Protection Elicited by a DNA Vaccine Candidate Expressing the prM-E Antigen of Dengue Virus Serotype 3 in Mice

“…The generation of both cell-mediated and NAb responses against DENV3 increased substantially (Versiani et al, 2017). Hurtado-Melgoza et al reported that the DNA vaccine candidate targeting DENV3 NS3 triggered a favorable response with the activation of T lymphocytes; however, immunization with this DNA vaccine presented no detectable antibody titers against DENV3 (Hurtado-Melgoza et al, 2016).…”

Commentary: Differential Humoral and Cellular Immunity Induced by Vaccination using Plasmid DNA and Protein Recombinant Expressing the NS3 Protein of Dengue Virus type 3

“…This response was of low magnitude, especially when compared to the recombinant protein vaccine in which antibody titers were 320 times higher. These results suggested that mice immunized with the gene coding for the NS3-DEN3 protein are primed to develop a Th1 response which provide protection against infection with dengue virus 12 .…”

“…Having a vaccine against the four strains of DENV which is safe, effective and affordable will be a breakthrough for disease control. This was one of the main triggers of the work of Hurtado-Melgoza et al 12 . First, we would have to consider that DENV infection induces potent innate immune responses, which in turn shape the adaptive immune responses, including not only B cell humoral responses with antibody production but also cellular responses mediated by CD4+ and CD8+ T cells 13 .…”

NS38 protein of GCRV 096: Functional analysis and effect on grass carp reovirus replication

“…Non-structural proteins play important role in viral infection, viral replication and proliferation, pathogenesis and immune evasion (Schierhorn et al, 2017). Some viral non-structural proteins have been considered as virulence factors (DeDiego et al, 2016), vaccine candidates (Hurtado-Melgoza et al, 2016;Kitagawa et al, 2017;Reyes-Sandoval and Ludert, 2019) and major targets for antiviral therapy (Chu et al, 2017). However, functions of GCRV non-structural proteins are not well understood.…”