Evaluation on the efficacy and immunogenicity of recombinant DNA plasmids expressing spike genes from porcine transmissible gastroenteritis virus and porcine epidemic diarrhea virus

PLoS One. 2013;8(3):e57468. doi: 10.1371/journal.pone.0057468. Epub 2013 Mar 19.

Abstract

Porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PDEV) can cause severe diarrhea in pigs. Development of effective vaccines against TGEV and PEDV is one of important prevention measures. The spike (S) protein is the surface glycoprotein of TGEV and PEDV, which can induce specific neutralization antibodies and is a candidate antigen for vaccination attempts. In this study, the open reading frames of the TGEV S1 protein and in addition of the S or S1 proteins of PEDV were inserted into the eukaryotic expression vector, pIRES, resulting in recombinant plasmids, pIRES-(TGEV-S1-PEDV-S1) and pIRES-(TGEV-S1-PEDV-S). Subsequently, 6-8 weeks old Kunming mice were inoculated with both DNA plasmids. Lymphocyte proliferation assay, virus neutralization assay, IFN-γ assay and CTL activity assay were performed. TGEV/PEDV specific antibody responses as well as kinetic changes of T lymphocyte subgroups of the immunized mice were analyzed. The results showed that the recombinant DNA plasmids increased the proliferation of T lymphocytes and the number of CD4+ and CD8+ T lymphocyte subgroups. In addition, the DNA vaccines induced a high level of IFN-γ in the immunized mice. The specific CTL activity in the pIRES-(TGEV-S1-PEDV-S) group became significant at 42 days post-immunization. At 35 days post-immunization, the recombinant DNA plasmids bearing full-length S genes of TGEV and PEDV stimulated higher levels of specific antibodies and neutralizing antibodies in immunized mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / blood
  • Coronavirus Infections / immunology
  • Coronavirus Infections / prevention & control
  • Coronavirus Infections / veterinary
  • DNA, Recombinant / genetics
  • DNA, Recombinant / immunology
  • DNA, Viral / genetics
  • DNA, Viral / immunology
  • Gastroenteritis, Transmissible, of Swine / immunology
  • Gastroenteritis, Transmissible, of Swine / prevention & control
  • Genes, Viral
  • Interferon-gamma / blood
  • Interleukin-4 / blood
  • Mice
  • Plasmids / genetics
  • Plasmids / immunology
  • Porcine epidemic diarrhea virus / genetics*
  • Porcine epidemic diarrhea virus / immunology*
  • Swine
  • Swine Diseases / immunology
  • Swine Diseases / prevention & control
  • T-Lymphocytes / immunology
  • Transmissible gastroenteritis virus / genetics*
  • Transmissible gastroenteritis virus / immunology*
  • Vaccines, DNA / genetics*
  • Vaccines, DNA / immunology*
  • Viral Proteins / genetics*
  • Viral Proteins / immunology*
  • Viral Vaccines / genetics*
  • Viral Vaccines / immunology*

Substances

  • Antibodies, Viral
  • DNA, Recombinant
  • DNA, Viral
  • Vaccines, DNA
  • Viral Proteins
  • Viral Vaccines
  • spike protein S, Transmissible gastroenteritis virus
  • Interleukin-4
  • Interferon-gamma

Grants and funding

The authors acknowledge National Natural Science Foundation of China (31270187), The Program for New Century Excellent Talents in University of Ministry of Education of P.R. China (NCET-10-0144), Sponsored by Chang Jiang Scholar Candidates Programme for Provincial Universities in Heilongjiang, Research Team Program on Scientific and Technological Innovation in Heilongjiang Provincial University (2011TD001), Innovation Talent Project (Excellent discipline leader) of the Harbin Science and Technology Bureau (RC2012XK002003). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.