Construction of a recombinant Lactococcus lactis strain expressing a fusion protein of Omp22 and HpaA from Helicobacter pylori for oral vaccine development

Biotechnol Lett. 2016 Nov;38(11):1911-1916. doi: 10.1007/s10529-016-2173-5. Epub 2016 Jul 12.

Abstract

Objectives: To develop orally administrated anti-Helicobacter pylori vaccination, a Lactococcus lactis strain was genetically constructed for fusion expression of H. pylori protective antigens HpaA and Omp22.

Results: The fusion gene of omp22 and hpaA with an adapter encoding three glycines was cloned from a plasmid pMAL-c2x-omp22-hpaA into Escherichia coli MC1061 and L. lactis NZ3900 successively using a shutter vector pNZ8110. Expression of the fusion gene in L. lactis was induced with nisin resulting in production of proteins with molecular weights of 50 and 28 kDa. Both of them were immunoreactive with mouse anti-H. pylori sera as determined via western blotting. Oral vaccination of BALB/c mice using the L. lactis strain carrying pNZ8110-omp22-hpaA elicited significant systematic humoral immune response (P < 0.05).

Conclusions: This is the first report showing that a fusion protein of two H. pylori antigens was efficiently expressed in L. lactis with immunogenicity. This is a considerable step towards H. pylori vaccines.

Keywords: Fusion protein; Helicobactor pylori; Lactococcus lactis; Oral vaccine; hpaA; omp22.

MeSH terms

  • Adhesins, Bacterial / genetics
  • Adhesins, Bacterial / immunology
  • Adhesins, Bacterial / metabolism*
  • Administration, Oral
  • Animals
  • Antibodies, Bacterial / blood
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology
  • Antigens, Bacterial / metabolism*
  • Bacterial Vaccines / immunology
  • Cloning, Molecular
  • Helicobacter pylori / genetics
  • Helicobacter pylori / immunology
  • Helicobacter pylori / metabolism*
  • Lactococcus lactis / genetics*
  • Lactococcus lactis / immunology
  • Lactococcus lactis / metabolism
  • Lipoproteins / genetics
  • Lipoproteins / immunology
  • Lipoproteins / metabolism*
  • Mice
  • Mice, Inbred BALB C
  • Nisin / pharmacology
  • Recombinant Fusion Proteins / administration & dosage
  • Recombinant Fusion Proteins / immunology*
  • Recombinant Fusion Proteins / metabolism
  • Vaccination

Substances

  • Adhesins, Bacterial
  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Bacterial Vaccines
  • Lipoproteins
  • Lpp20 protein, Helicobacter pylori
  • Recombinant Fusion Proteins
  • Nisin
  • adhesin A, Helicobacter