OmpW expressed by recombinant Lactobacillus casei elicits protective immunity against Aeromonas veronii in common carp

Microb Pathog. 2019 Aug:133:103552. doi: 10.1016/j.micpath.2019.103552. Epub 2019 May 20.

Abstract

Aeromonas veronii is an opportunistic pathogen that is capable of infecting both aquatic livestock and mammals. Natural infection in fishes results in irreparable damage to the aquaculture industry. In this study, we sought to investigate whether recombinant Lactobacillus casei expressing the outer membrane protein W (OmpW) of A.veronii could elicit protective immunity against A.veronii infections. We generated two recombinant Lactobacillus casei (L.casei) strains expressing the OmpW of A.veronii (surface-displayed or secreted) and evaluated the effect on immune responses in a fish model. A 600-bp gene fragment was subcloned into the L.casei expression plasmids pPG-1 (surface-displayed) and pPG-2 (secreted). Expression of the recombinant OmpW protein was also confirmed by Western blot and immunofluorescence assays. Common carp immunized with Lc-pPG-1- OmpW and Lc-pPG-2- OmpW via oral administration elicited high serum specific antibody titers and high LZM, ACP, and SOD activities. High levels of the IL-10, IL-β, IFN-γ, and TNF-α genes in different organs indicated that the inflammatory response and cell immune response were triggered. Additionally, when immunized fish were challenged with A.veronii, Lc-pPG1-OmpW and Lc-pPG2-OmpW demonstrated 40% and 50% protective efficacy. These data indicate that the combination of OmpW delivery and the lactic acid bacteria (LAB) approach may be a promising mucosal therapeutic strategy for treatment of A.veronii.

Keywords: Aeromonas veronii; Immune protection; Lactobacillus casei; OmpW.

MeSH terms

  • Administration, Oral
  • Aeromonas veronii / genetics*
  • Aeromonas veronii / immunology
  • Animals
  • Antibodies, Bacterial / blood
  • Bacterial Outer Membrane Proteins / administration & dosage
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / immunology*
  • Bacterial Vaccines
  • Base Sequence
  • Carps / immunology*
  • Carps / microbiology
  • Cytokines / genetics
  • Cytokines / metabolism
  • Disease Models, Animal
  • Fish Diseases / immunology
  • Fish Diseases / prevention & control*
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial / genetics
  • Gram-Negative Bacterial Infections / immunology
  • Gram-Negative Bacterial Infections / prevention & control*
  • Gram-Negative Bacterial Infections / veterinary*
  • Immunity, Cellular
  • Immunity, Humoral
  • Immunization / veterinary*
  • Interferon-gamma / genetics
  • Interleukin-10 / genetics
  • Intestines / microbiology
  • Lacticaseibacillus casei / genetics
  • Lacticaseibacillus casei / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Tumor Necrosis Factor-alpha / genetics

Substances

  • Antibodies, Bacterial
  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Cytokines
  • Recombinant Proteins
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • Interferon-gamma