Delivery of Helicobacter pylori HpaA to gastrointestinal mucosal immune sites using Lactococcus lactis and its immune efficacy in mice

Biotechnol Lett. 2018 Mar;40(3):585-590. doi: 10.1007/s10529-017-2502-3. Epub 2018 Jan 3.

Abstract

Objective: To develop a safe and effective oral vaccine against Helicobacter pylori using its HpaA protein expressed in Lactococcus lactis.

Results: The gene encoding HpaA was obtained by PCR and ligated to pNZ8110-lysM following digestion with NaeI + SphI. The recombinant plasmid was transferred into E. coli for multiplication, and then into L. lactis. The recombinant L. lactis was induced to express HpaA, resulting in two products of 29 and 25 kDa, both of which yielded positive immunoreaction with mouse antisera against H. pylori, as confirmed by immunoblot assays. The 29 kDa product constituted 12% of the cell lysates. Oral inoculation with the engineered L. lactis evoked significantly elevated serum IgG level in mice (P < 0.05).

Conclusions: A novel engineered L. lactis strain was developed that efficiently produces whole HpaA protein with desired antigenicity and potent immunogenicity. It provides a basis for approaches to L. lactis-delivered anti-H. pylori vaccination.

Keywords: Adhesin; Genetic engineering; Helicobacter pylori; Lactococcus lactis; Mucosal immune.

MeSH terms

  • Adhesins, Bacterial / chemistry
  • Adhesins, Bacterial / genetics
  • Adhesins, Bacterial / immunology*
  • Adhesins, Bacterial / metabolism
  • Animals
  • Antibodies, Bacterial / blood*
  • Antibodies, Bacterial / immunology
  • Bacterial Vaccines / chemistry
  • Bacterial Vaccines / genetics
  • Bacterial Vaccines / immunology*
  • Bacterial Vaccines / metabolism
  • Cloning, Molecular
  • Genetic Engineering
  • Helicobacter Infections / immunology
  • Helicobacter Infections / prevention & control
  • Helicobacter pylori / genetics
  • Helicobacter pylori / immunology*
  • Immunity, Mucosal / immunology
  • Lactococcus lactis / genetics
  • Lactococcus lactis / metabolism*
  • Mice
  • Mice, Inbred BALB C
  • Probiotics
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology*
  • Recombinant Fusion Proteins / metabolism

Substances

  • Adhesins, Bacterial
  • Antibodies, Bacterial
  • Bacterial Vaccines
  • Recombinant Fusion Proteins
  • adhesin A, Helicobacter