A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice

J Virol Methods. 2015 Dec 1:225:35-40. doi: 10.1016/j.jviromet.2015.09.001. Epub 2015 Sep 5.

Abstract

Peste des petits ruminants (PPR), a highly contagious disease induced by PPR virus (PPRV), affects sheep and goats. PPRV fusion (F) protein is important for the induction of immune responses against PPRV. We constructed a Semliki Forest virus (SFV) replicon-vectored DNA vaccine ("suicidal DNA vaccine") and evaluated its immunogenicity in BALB/c mice. The F gene of PPRV was cloned and inserted into the SFV replicon-based vector pSCA1. The antigenicity of the resultant plasmid pSCA1/F was identified by indirect immunofluorescence and western blotting. BALB/c mice were then intramuscularly injected with pSCA1/F three times at 14-d intervals. Specific antibodies and virus-neutralizing antibodies against PPRV were quantified by indirect ELISA and microneutralization tests, respectively. Cell-mediated immune responses were examined by cytokine and lymphocyte proliferation assays. The pSCA1/F expressed F protein in vitro and induced specific and neutralizing antibody production, and lymphocyte proliferation in mice. Mice vaccinated with pSCA1/F had increased IL-2 and IL-10 levels after 24-h post first immunization. IFN-γ and TNF-α levels increased from that time point and gradually decreased thereafter. Thus, the Semliki Forest virus replicon-vectored DNA vaccine expressing the F protein of PPRV induced both humoral and cell-mediated immune responses in mice. This could be considered as a novel strategy for vaccine development against PPR.

Keywords: Fusion protein; Immune response; Peste des petits ruminants virus; Suicidal DNA vaccine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Viral / blood*
  • Cell Proliferation
  • Cloning, Molecular
  • Cytokines / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Immunization Schedule
  • Injections, Intramuscular
  • Leukocytes, Mononuclear / immunology*
  • Mice, Inbred BALB C
  • Neutralization Tests
  • Peste-des-petits-ruminants virus / genetics
  • Peste-des-petits-ruminants virus / immunology*
  • Plasmids
  • Semliki forest virus / genetics
  • Vaccines, DNA / administration & dosage
  • Vaccines, DNA / genetics
  • Vaccines, DNA / immunology*
  • Viral Fusion Proteins / genetics
  • Viral Fusion Proteins / immunology*

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Cytokines
  • F protein, Peste-des-petits-ruminants virus
  • Vaccines, DNA
  • Viral Fusion Proteins