Oral administration of Lactococcus lactis expressing Helicobacter pylori Cag7-ct383 protein induces systemic anti-Cag7 immune response in mice

FEMS Immunol Med Microbiol. 2009 Dec;57(3):257-68. doi: 10.1111/j.1574-695X.2009.00605.x. Epub 2009 Sep 1.

Abstract

To express the 3'-region (1152 bp) of the cag7 gene of Helicobacter pylori 51 strain, encoding the C-terminal 383 amino acid (ct383 aa) region of Cag7 protein that is known to cover the needle region of T4SS, in a live delivery vehicle Lactococcus lactis, the cag7-ct383 gene was amplified by PCR. DNA sequence analysis revealed that the amino acid sequence of Cag7-ct383 of H. pylori 51 shared 98.4% and 97.4% identity with H. pylori 26695 and J99, respectively. Intramuscular injection of the GST-Cag7-ct383 fusion protein into a rat could raise the anti-Cag7 antibody, indicating the immunogenicity of the Cag7-ct383 protein. When the cag7-ct383 gene was cloned in Escherichia coli-L. lactis shuttle vector (pMG36e) and transformed into L. lactis, the transformant could produce the Cag7-ct383 protein, as evidenced by Western blot analysis. The Cag7-ct383 protein level in the L. lactis transformant reached a maximum at the early stationary phase without extracellular secretion. The oral administration of the L. lactis transformant into mice generated anti-Cag7 antibody in serum in five of five mice. These results suggest that L. lactis transformant expressing Cag7-ct383 protein may be applicable as an oral vaccine to induce mucosal and systemic immunity to H. pylori.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Oral
  • Amino Acid Sequence
  • Animals
  • Antibodies, Bacterial / blood
  • Bacterial Proteins / genetics
  • Bacterial Proteins / immunology*
  • Bacterial Vaccines / administration & dosage
  • Bacterial Vaccines / genetics
  • Bacterial Vaccines / immunology*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Genetic Vectors*
  • Helicobacter Infections / immunology
  • Helicobacter Infections / prevention & control*
  • Helicobacter pylori / genetics
  • Helicobacter pylori / immunology*
  • Lactococcus lactis / genetics*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods
  • Rats
  • Rats, Sprague-Dawley
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology

Substances

  • Antibodies, Bacterial
  • Bacterial Proteins
  • Bacterial Vaccines
  • CAG7 protein, Helicobacter pylori
  • DNA, Bacterial
  • Vaccines, Synthetic