Molecular mechanisms underlying protection against H9N2 influenza virus challenge in mice by recombinant Lactobacillus plantarum with surface displayed HA2-LTB

J Biotechnol. 2017 Oct 10:259:6-14. doi: 10.1016/j.jbiotec.2017.08.011. Epub 2017 Aug 12.

Abstract

It has been considered that the Avian influenza virus (AIV) causes severe threats to poultry industry. In this study, we constructed a series of recombinant Lactobacillus plantarum (L. plantarum) with surface displayed hemagglutinin subunit 2 (HA2) alone or together with heat-labile toxin B subunit (LTB) from enterotoxigenic Escherichia coli. Balb/c mice were used as model to evaluate the protective effects of recombinant L. plantarum strains against H9N2 subtype challenge. The results showed that the presence of LTB significantly increased the percentages of CD3+CD4+IL-4+, CD3+CD4+IFN-γ+ and CD3+CD4+IL-17+ T cells, as well as CD3+CD8+IFN-γ+ T cells in spleen and MLNs determined by Fluorescence-Activated Cell Sorting assay. Similar increased production of serum IFN-γ was also confirmed by enzyme linked immunosorbent assay (ELISA). The L. plantarum with surface displayed HA2-LTB also dramatically increased the percentages of B220+ IgA+ B cells in peyer patch, in consistent with elevated production of mucosal SIgA antibody determined by ELISA. Finally, the orally administrated HA2-LTB expressing strain efficiently protected mice against H9N2 subtype AIV challenge shown by increased survival percentages, body weight gains and decreased lung lesions in histopathologic analysis. In conclusion, this study provides more detail mechanisms underlying the adjuvant effects of LTB on heterologous antigen produced in recombinant lactic acid bacteria.

Keywords: AIV; HA2 protein; Heat-labile toxin B subunit (LTB); Immune response; L. plantarum.

MeSH terms

  • Animals
  • Antibodies, Viral
  • Bacterial Toxins / genetics*
  • Cell Surface Display Techniques
  • Cytokines / blood
  • Escherichia coli
  • Hemagglutinins, Viral* / genetics
  • Hemagglutinins, Viral* / immunology
  • Influenza A Virus, H9N2 Subtype / genetics*
  • Influenza Vaccines* / genetics
  • Influenza Vaccines* / immunology
  • Lactobacillus plantarum* / genetics
  • Lactobacillus plantarum* / immunology
  • Lactobacillus plantarum* / metabolism
  • Mice
  • Orthomyxoviridae Infections / prevention & control*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology
  • Vaccines, Synthetic / metabolism

Substances

  • Antibodies, Viral
  • Bacterial Toxins
  • Cytokines
  • Hemagglutinins, Viral
  • Influenza Vaccines
  • Recombinant Fusion Proteins
  • Vaccines, Synthetic