Oral Immunization of Chickens with Lactococcus lactis Expressing cjaA Temporarily Reduces Campylobacter jejuni Colonization

Foodborne Pathog Dis. 2020 Jun;17(6):366-372. doi: 10.1089/fpd.2019.2727. Epub 2019 Nov 13.

Abstract

Campylobacter jejuni is the leading cause of human foodborne enteritis worldwide. Poultry products are regarded as the main source of human campylobacteriosis. Strategies are being developed to reduce colonization of poultry by Campylobacter. The membrane transport protein CjaA was reported to stimulate mucosal immune responses, which can reduce the C. jejuni load in chickens. In this study, oral immunization of broilers with food-grade Lactococcus lactis NZ3900/pNZ8149 carrying the C. jejuni cjaA gene was examined for the ability to reduce colonization of broilers by Campylobacter. The Usp45 signal peptide and the Escherichia coli heat-labile enterotoxin B subunit (LTB) gene fragments were inserted into the upstream and downstream of the cjaA gene for secretory expression and immune enhancement, respectively. The cjaA gene and the fusion cjaA-ltb gene were both expressed in recombinant L. lactis, and the single cjaA gene was secretory expressed in the recombinant strain. Oral administration of two recombinant L. lactis strains expressing the cjaA gene and the fusion cjaA-ltb gene both stimulated specific anti-CjaA serum IgY responses significantly. While the average intestinal sIgA responses in these groups were higher compared with the control groups, they were not significantly different. Chicken challenge experiments showed that the colonization levels of C. jejuni in the groups provided oral immunization with two recombinant L. lactis-delivered CjaA strains were significantly lower than that of the control group at 5 d postinoculation, but there was no significant difference in C. jejuni colonization among all groups at 9 d. These results indicated that recombinant L. lactis with secretory expression of CjaA is a promising live vector vaccine against C. jejuni colonization of chickens. The immunization regimen requires further optimization to ideally stimulate detectable levels of intestinal sIgA to enhance the level of inhibition of C. jejuni colonization.

Keywords: Campylobacter jejuni; Lactococcus lactis; membrane transport protein CjaA; oral immunization; secretory expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / genetics
  • ATP-Binding Cassette Transporters / immunology*
  • Administration, Oral
  • Amino Acid Transport Systems, Neutral / genetics
  • Amino Acid Transport Systems, Neutral / immunology*
  • Animals
  • Bacterial Toxins / genetics
  • Bacterial Toxins / immunology
  • Bacterial Vaccines
  • Campylobacter Infections / prevention & control*
  • Campylobacter Infections / veterinary
  • Campylobacter jejuni / growth & development*
  • Chickens
  • Enterotoxins / genetics
  • Enterotoxins / immunology
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / immunology
  • Humans
  • Immunization / methods
  • Immunization / veterinary
  • Lactococcus lactis*
  • Membrane Transport Proteins / genetics
  • Membrane Transport Proteins / immunology
  • Poultry Diseases / prevention & control
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Specific Pathogen-Free Organisms

Substances

  • ATP-Binding Cassette Transporters
  • Amino Acid Transport Systems, Neutral
  • Bacterial Toxins
  • Bacterial Vaccines
  • CjaA protein, Campylobacter jejuni
  • Enterotoxins
  • Escherichia coli Proteins
  • Membrane Transport Proteins
  • Recombinant Proteins
  • heat-labile enterotoxin, E coli